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Sample records for piasy-mediated sumo modification

  1. Regulation of SUMO Modification

    NARCIS (Netherlands)

    P.M. Knipscheer (Puck Maria)

    2007-01-01

    textabstractThe small ubiquitin related modifier SUMO is a posttranslational modifier that functions in a wide range of cellular processes like intracellular transport, cell cycle regulation, DNA repair and regulation of transcription. SUMO is an 11 kDa protein and is ligated to its target proteins

  2. Regulation of REGγ cellular distribution and function by SUMO modification

    Institute of Scientific and Technical Information of China (English)

    Yan Wu; Honglin Luo; Xiaotao Li; Lu Wang; Ping Zhou; Guangqiang Wang; Yu Zeng; Ying Wang; Jian Liu; Bianhong Zhang; Shuang Liu

    2011-01-01

    Discovery of emerging REGy-regulated proteins has accentuated the RECry-proteasome as an important pathway in multiple biological processes, including cell growth, cell cycle regulation, and apoptosis. However, little is known about the regulation of the REGy-proteasome pathway. Here we demonstrate that REGγ can be SUMOylated in vitro and in vivo by SUMO-1, SUMO-2, and SUMO-3. The SUMO-E3 protein inhibitor of activated STAT(PIAS)1physically associates with REGy and promotes SUMOylation of REGy. SUMOylation of RECry was found to occur at multiple sites, including K6, K14, and K12. Mutation analysis indicated that these SUMO sites simultaneously contributed to the SUMOylation status of REGy in cells. Posttranslational modification of REGγ by SUMO conjugation was revealed to mediate cytosolic translocation of REGγ and to cause increased stability of this proteasome activator.SUMOylation-deficient REGγ displayed attenuated ability to degrade p21waf//Cipl due to reduced affinity of the REGγ SUMOylation-defective mutant for p21. Taken together, we report a previously unrecognized mechanism regulating the activity of the proteasome activator REGy. This regulatory mechanism may enable REGy to function as a more potent factor in protein degradation with a broader substrate spectrum.

  3. Principles of ubiquitin and SUMO modifications in DNA repair

    NARCIS (Netherlands)

    Bergink, Steven; Jentsch, Stefan

    2009-01-01

    With the discovery in the late 1980s that the DNA-repair gene RAD6 encodes a ubiquitin-conjugating enzyme, it became clear that protein modification by ubiquitin conjugation has a much broader significance than had previously been assumed. Now, two decades later, ubiquitin and its cousin SUMO are im

  4. Structure of the Siz/PIAS SUMO E3 Ligase Siz1 and Determinants Required for SUMO Modification of PCNA

    Energy Technology Data Exchange (ETDEWEB)

    Yunus, Ali A.; Lima, Christopher D.; (SKI)

    2010-01-12

    Siz1 is a founding member of the Siz/PIAS RING family of SUMO E3 ligases. The X-ray structure of an active Siz1 ligase revealed an elongated tripartite architecture comprised of an N-terminal PINIT domain, a central zinc-containing RING-like SP-RING domain, and a C-terminal domain we term the SP-CTD. Structure-based mutational analysis and biochemical studies show that the SP-RING and SP-CTD are required for activation of the E2SUMO thioester, while the PINIT domain is essential for redirecting SUMO conjugation to the proliferating cell nuclear antigen (PCNA) at lysine 164, a nonconsensus lysine residue that is not modified by the SUMO E2 in the absence of Siz1. Mutational analysis of Siz1 and PCNA revealed surfaces on both proteins that are required for efficient SUMO modification of PCNA in vitro and in vivo.

  5. A Critical SUMO1 Modification of LKB1 Regulates AMPK Activity during Energy Stress

    KAUST Repository

    Ritho, Joan

    2015-07-23

    SUMOylation has been implicated in cellular stress adaptation, but its role in regulating liver kinase B1 (LKB1), a major upstream kinase of the energy sensor AMP-activated protein kinase (AMPK), is unknown. Here, we show that energy stress triggers an increase in SUMO1 modification of LKB1, despite a global reduction in both SUMO1 and SUMO2/3 conjugates. During metabolic stress, SUMO1 modification of LKB1 lysine 178 is essential in promoting its interaction with AMPK via a SUMO-interacting motif (SIM) essential for AMPK activation. The LKB1 K178R SUMO mutant had defective AMPK signaling and mitochondrial function, inducing death in energy-deprived cells. These results provide additional insight into how LKB1-AMPK signaling is regulated during energy stress, and they highlight the critical role of SUMOylation in maintaining the cell’s energy equilibrium.

  6. Modification of Akt by SUMO conjugation regulates alternative splicing and cell cycle

    OpenAIRE

    Risso, Guillermo; Pelisch, Federico; Pozzi, Berta; Mammi, Pablo; Blaustein, Matías; Colman-Lerner, Alejandro; Srebrow, Anabella

    2013-01-01

    Akt/PKB is a key signaling molecule in higher eukaryotes and a crucial protein kinase in human health and disease. Phosphorylation, acetylation, and ubiquitylation have been reported as important regulatory post-translational modifications of this kinase. We describe here that Akt is modified by SUMO conjugation, and show that lysine residues 276 and 301 are the major SUMO attachment sites within this protein. We found that phosphorylation and SUMOylation of Akt appear as independent events. ...

  7. Divergent signaling via SUMO modification: potential for CFTR modulation.

    Science.gov (United States)

    Ahner, Annette; Gong, Xiaoyan; Frizzell, Raymond A

    2016-02-01

    The cystic fibrosis transmembrane conductance regulator (CFTR) is generally responsible for the cAMP/PKA regulated anion conductance at the apical membranes of secretory epithelial cells. Mutations in CFTR underlie cystic fibrosis (CF), in which the most common variant, F508del, causes protein misfolding and its proteasome-mediated degradation. A new pathway that contributes to mutant CFTR degradation is mediated by the small heat shock protein, Hsp27, which cooperates with Ubc9, the E2 enzyme for SUMOylation, to selectively conjugate mutant CFTR with SUMO-2/3. This SUMO paralog can form polychains, which are recognized by the ubiquitin E3 enzyme, RNF4, leading to CFTR ubiquitylation and recognition by the proteasome. We found also that F508del CFTR could be modified by SUMO-1, a paralog that does not support SUMO polychain formation. The use of different SUMO paralogs to modify and target a single substrate for divergent purposes is not uncommon. In this short review we discuss the possibility that conjugation with SUMO-1 could protect mutant CFTR from disposal by RNF4 and similar ubiquitin ligases. We hypothesize that such a pathway could contribute to therapeutic efforts to stabilize immature mutant CFTR and thereby enhance the action of therapeutics that correct CFTR trafficking to the apical membranes.

  8. KSHV latent protein LANA2 inhibits sumo2 modification of p53

    Science.gov (United States)

    Laura, Marcos-Villar; de la Cruz-Herrera, Carlos F; Ferreirós, Alba; Baz-Martínez, Maite; Lang, Valerie; Vidal, Anxo; Muñoz-Fontela, Cesar; Rodríguez, Manuel S; Collado, Manuel; Rivas, Carmen

    2015-01-01

    Tumor suppressor p53 plays a crucial antiviral role and targeting of p53 by viral proteins is a common mechanism involved in virus oncogenesis. The activity of p53 is tightly regulated at the post-translational levels through a myriad of modifications. Among them, modification of p53 by SUMO has been associated with the onset of cellular senescence. Kaposi´s sarcoma-associated herpesvirus (KSHV) expresses several proteins targeting p53, including the latent protein LANA2 that regulates polyubiquitylation and phosphorylation of p53. Here we show that LANA2 also inhibits the modification of p53 by SUMO2. Furthermore, we show that the reduction of p53-SUMO2 conjugation by LANA2, as well as the p53-LANA2 interaction, both require the SUMOylation of the viral protein and its interaction with SUMO or SUMOylated proteins in a non-covalent manner. Finally, we show that the control of p53-SUMO2 conjugation by LANA2 correlates with its ability to inhibit SUMO2- and type I interferon-induced senescence. These results highlight the importance of p53 SUMOylation in the control of virus infection and suggest that viral oncoproteins could contribute to viral infection and cell transformation by abrogating p53 SUMOylation. PMID:25607652

  9. Analysis of Changes in SUMO-2/3 Modification during Breast Cancer Progression and Metastasis

    DEFF Research Database (Denmark)

    Subramonian, Divya; Raghunayakula, Sarita; Olsen, Jesper V;

    2014-01-01

    SUMOylation is an essential posttranslational modification and regulates many cellular processes. Dysregulation of SUMOylation plays a critical role in metastasis, yet how its perturbation affects this lethal process of cancer is not well understood. We found that SUMO-2/3 modification is greatly...

  10. Characterization of a novel posttranslational modification in polypyrimidine tract-binding proteins by SUMO1.

    Science.gov (United States)

    Han, Wei; Wang, Lin; Yin, Bin; Peng, Xiaozhong

    2014-04-01

    Polypyrimidine tract-binding protein 1 (PTBP1) and its brainspecific homologue, PTBP2, are associated with pre-mRNAs and influence pre-mRNA processing, as well as mRNA metabolism and transport. They play important roles in neural differentiation and glioma development. In our study, we detected the expression of the two proteins in glioma cells and predicted that they may be sumoylated using SUMOplot analyses. We confirmed that PTBP1 and PTBP2 can be modified by SUMO1 with co-immunoprecipitation experiments using 293ET cells transiently co-expressing SUMO1 and either PTBP1 or PTBP2. We also found that SUMO1 modification of PTBP2 was enhanced by Ubc9 (E2). The mutation of the sumoylation site (Lys137) of PTBP2 markedly inhibited its modification by SUMO1. Interestingly, in T98G glioma cells, the level of sumoylated PTBP2 was reduced compared to that of normal brain cells. Overall, this study shows that PTBP2 is posttranslationally modified by SUMO1.

  11. SUMO modification regulates BLM and RAD51 interaction at damaged replication forks.

    Directory of Open Access Journals (Sweden)

    Karen J Ouyang

    2009-12-01

    Full Text Available The gene mutated in Bloom's syndrome, BLM, is important in the repair of damaged replication forks, and it has both pro- and anti-recombinogenic roles in homologous recombination (HR. At damaged forks, BLM interacts with RAD51 recombinase, the essential enzyme in HR that catalyzes homology-dependent strand invasion. We have previously shown that defects in BLM modification by the small ubiquitin-related modifier (SUMO cause increased gamma-H2AX foci. Because the increased gamma-H2AX could result from defective repair of spontaneous DNA damage, we hypothesized that SUMO modification regulates BLM's function in HR repair at damaged forks. To test this hypothesis, we treated cells that stably expressed a normal BLM (BLM+ or a SUMO-mutant BLM (SM-BLM with hydroxyurea (HU and examined the effects of stalled replication forks on RAD51 and its DNA repair functions. HU treatment generated excess gamma-H2AX in SM-BLM compared to BLM+ cells, consistent with a defect in replication-fork repair. SM-BLM cells accumulated increased numbers of DNA breaks and were hypersensitive to DNA damage. Importantly, HU treatment failed to induce sister-chromatid exchanges in SM-BLM cells compared to BLM+ cells, indicating a specific defect in HR repair and suggesting that RAD51 function could be compromised. Consistent with this hypothesis, RAD51 localization to HU-induced repair foci was impaired in SM-BLM cells. These data suggested that RAD51 might interact noncovalently with SUMO. We found that in vitro RAD51 interacts noncovalently with SUMO and that it interacts more efficiently with SUMO-modified BLM compared to unmodified BLM. These data suggest that SUMOylation controls the switch between BLM's pro- and anti-recombinogenic roles in HR. In the absence of BLM SUMOylation, BLM perturbs RAD51 localization at damaged replication forks and inhibits fork repair by HR. Conversely, BLM SUMOylation relieves its inhibitory effects on HR, and it promotes RAD51 function.

  12. SIZ1-Dependent Post-Translational Modification by SUMO Modulates Sugar Signaling and Metabolism in Arabidopsis thaliana.

    Science.gov (United States)

    Castro, Pedro Humberto; Verde, Nuno; Lourenço, Tiago; Magalhães, Alexandre Papadopoulos; Tavares, Rui Manuel; Bejarano, Eduardo Rodríguez; Azevedo, Herlânder

    2015-12-01

    Post-translational modification mechanisms function as switches that mediate the balance between optimum growth and the response to environmental stimuli, by regulating the activity of key proteins. SUMO (small ubiquitin-like modifier) attachment, or sumoylation, is a post-translational modification that is essential for the plant stress response, also modulating hormonal circuits to co-ordinate developmental processes. The Arabidopsis SUMO E3 ligase SAP and Miz 1 (SIZ1) is the major SUMO conjugation enhancer in response to stress, and is implicated in several aspects of plant development. Here we report that known SUMO targets are over-represented in multiple carbohydrate-related proteins, suggesting a functional link between sumoylation and sugar metabolism and signaling in plants. We subsequently observed that SUMO-conjugated proteins accumulate in response to high doses of sugar in a SIZ1-dependent manner, and that the null siz1 mutant displays increased expression of sucrose and starch catabolic genes and shows reduced starch levels. We demonstrated that SIZ1 controls germination time and post-germination growth via osmotic and sugar-dependent signaling, respectively. Glucose was specifically linked to SUMO-sugar interplay, with high levels inducing root growth inhibition and aberrant root hair morphology in siz1. The use of sugar analogs and sugar marker gene expression analysis allowed us to implicate SIZ1 in a signaling pathway dependent on glucose metabolism, probably involving modulation of SNF1-related kinase 1 (SnRK1) activity.

  13. Principles of protein group SUMO modification substantiated in DNA repair

    OpenAIRE

    Psakhye, Ivan

    2013-01-01

    Posttranslational modifications (PTMs) of proteins by covalent attachment of functional groups (like phosphorylation, acetylation, methylation, glycosylation, etc.) are of key importance for the cell as they regulate various aspects of protein behavior after its synthesis, e.g., dictate protein interaction properties, change catalytic activity of enzymes, induce conformational changes, guide subcellular localization and determine protein stability. A special class of protein PTMs is the conju...

  14. Small ubiquitin-like modifier (SUMO) modification of zinc finger protein 131 potentiates its negative effect on estrogen signaling.

    Science.gov (United States)

    Oh, Yohan; Chung, Kwang Chul

    2012-05-18

    Like ubiquitin, small ubiquitin-like modifier (SUMO) covalently attaches to specific target proteins and modulates their functional properties, including subcellular localization, protein dimerization, DNA binding, and transactivation of transcription factors. Diverse transcriptional co-regulator complexes regulate the ability of estrogen receptors to respond to positive and negative acting hormones. Zinc finger protein 131 (ZNF131) is poorly characterized but may act as a repressor of estrogen receptor α (ERα)-mediated trans-activation. Here, we identify ZNF131 as a target for SUMO modification and as a substrate for the SUMO E3 ligase human polycomb protein 2 (hPc2). We report that the SUMO-interacting motif 1 (SIM1) and the C-box of hPc2 are critical regions required for ZNF131 SUMOylation and define the ZNF131 SUMOylation site as lysine 567. We further show that SUMO modification potentiates the negative effect of ZNF131 on estrogen signaling and consequently attenuates estrogen-induced cell growth in a breast cancer cell line. Our findings suggest that SUMOylation is a novel regulator of ZNF131 action in estrogen signaling and breast cancer cell proliferation.

  15. The Smc5-Smc6 complex and SUMO modification of Rad52 regulates recombinational repair at the ribosomal gene locus

    DEFF Research Database (Denmark)

    Torres-Rosell, Jordi; Sunjevaric, Ivana; De Piccoli, Giacomo;

    2007-01-01

    at an extranucleolar site. The nucleolar exclusion of Rad52 recombination foci entails Mre11 and Smc5-Smc6 complexes and depends on Rad52 SUMO (small ubiquitin-related modifier) modification. Remarkably, mutations that abrogate these activities result in the formation of Rad52 foci within the nucleolus and cause r...

  16. SUMO: regulating the regulator

    Directory of Open Access Journals (Sweden)

    Bossis Guillaume

    2006-06-01

    Full Text Available Abstract Post-translational modifiers of the SUMO (Small Ubiquitin-related Modifier family have emerged as key regulators of protein function and fate. While the past few years have seen an enormous increase in knowledge on SUMO enzymes, substrates, and consequences of modification, regulation of SUMO conjugation is far from being understood. This brief review will provide an overview on recent advances concerning (i the interplay between sumoylation and other post-translational modifications at the level of individual targets and (ii global regulation of SUMO conjugation and deconjugation.

  17. Post-Translational Modifications of Kaposi’s Sarcoma-Associated Herpesvirus (KSHV Regulatory Proteins-SUMO and KSHV

    Directory of Open Access Journals (Sweden)

    Mel eCampbell

    2012-02-01

    Full Text Available Reactivation from a latent state is an important feature of infection and disease caused by many herpesviruses. KSHV latency can be envisioned as an outcome that is balanced between factors that promote viral gene expression and lytic replication against those that facilitate gene silencing and establish or maintain latency. A large body of work has focused on the activities of the key viral regulatory proteins involved in KSHV latent or lytic states. Moreover, recent studies have also begun to document the importance of epigenetic landscape evolution of the KSHV viral genome during latency and reactivation. However, one area of KSHV molecular virology that remains largely unanswered is the precise role of post-translational modifications on the activities of viral factors that function during latency and reactivation. In this review, we will summarize the post-translational modifications associated with three viral factors whose activities contribute to the viral state. The viral proteins discussed are the two major KSHV encoded transcription factors, K-Rta and K-bZIP (KSHV basic leucine zipper and the viral latency-associated nuclear antigen (LANA. A special emphasis will be placed on the role of the sumoylation pathway in the modulation of the KSHV lifecycle. Newly uncovered SUMO-dependent properties of LANA and K-Rta will also be presented, namely LANA histone-targeting SUMO E3 ligase activity and K-Rta SUMO-targeted ubiquitin ligase function.

  18. DNA Damage Response and SUMO Modification%SUMO化修饰在DNA损伤响应中的作用

    Institute of Scientific and Technical Information of China (English)

    蒋华东; 李钰

    2012-01-01

    物理或化学等多种因素均可以引起DNA损伤.为维持机体基因组的稳定性,机体形成了精确完整的机制来修复损伤的DNA. SUMO(small ubiquitin-related modifier,SUMO)化修饰与其他蛋白翻译后修饰一样,具有多种生物学功能.近年来的研究表明,其在DNA损伤修复中也具有非常重要的作用.该文就DNA损伤修复、SUMO化修饰系统及其二者关系的最新研究进展作了较为全面的介绍和总结.%DNA damage can be produced widely in our body by physical and/or chemical factors. Therefore, in order to maintain the genomic integrity, DNA must be protected from DNA damage and repaired correctly by forming accurate mechanism if DNA damage happened. SUMOylation has multiple biological functions like other post-translation modification. Emerging evidence has showed that SUMOs play vital roles in DNA damage repair. This review focuses on the newly research about DNA damage, SUMOylation and the relationship between them and provides a complete introducing and summarization.

  19. SUMO and KSHV Replication

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Pei-Ching [Institute of Microbiology and Immunology, National Yang-Ming University, Taipei 112, Taiwan (China); Kung, Hsing-Jien, E-mail: hkung@nhri.org.tw [Institute for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei 110, Taiwan (China); Department of Biochemistry and Molecular Medicine, University of California, Davis, CA 95616 (United States); UC Davis Cancer Center, University of California, Davis, CA 95616 (United States); Division of Molecular and Genomic Medicine, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli County 35053, Taiwan (China)

    2014-09-29

    Small Ubiquitin-related MOdifier (SUMO) modification was initially identified as a reversible post-translational modification that affects the regulation of diverse cellular processes, including signal transduction, protein trafficking, chromosome segregation, and DNA repair. Increasing evidence suggests that the SUMO system also plays an important role in regulating chromatin organization and transcription. It is thus not surprising that double-stranded DNA viruses, such as Kaposi’s sarcoma-associated herpesvirus (KSHV), have exploited SUMO modification as a means of modulating viral chromatin remodeling during the latent-lytic switch. In addition, SUMO regulation allows the disassembly and assembly of promyelocytic leukemia protein-nuclear bodies (PML-NBs), an intrinsic antiviral host defense, during the viral replication cycle. Overcoming PML-NB-mediated cellular intrinsic immunity is essential to allow the initial transcription and replication of the herpesvirus genome after de novo infection. As a consequence, KSHV has evolved a way as to produce multiple SUMO regulatory viral proteins to modulate the cellular SUMO environment in a dynamic way during its life cycle. Remarkably, KSHV encodes one gene product (K-bZIP) with SUMO-ligase activities and one gene product (K-Rta) that exhibits SUMO-targeting ubiquitin ligase (STUbL) activity. In addition, at least two viral products are sumoylated that have functional importance. Furthermore, sumoylation can be modulated by other viral gene products, such as the viral protein kinase Orf36. Interference with the sumoylation of specific viral targets represents a potential therapeutic strategy when treating KSHV, as well as other oncogenic herpesviruses. Here, we summarize the different ways KSHV exploits and manipulates the cellular SUMO system and explore the multi-faceted functions of SUMO during KSHV’s life cycle and pathogenesis.

  20. SUMO modification of Stra13 is required for repression of cyclin D1 expression and cellular growth arrest.

    Directory of Open Access Journals (Sweden)

    Yaju Wang

    Full Text Available Stra13, a basic helix-loop-helix (bHLH transcription factor is involved in myriad biological functions including cellular growth arrest, differentiation and senescence. However, the mechanisms by which its transcriptional activity and function are regulated remain unclear. In this study, we provide evidence that post-translational modification of Stra13 by Small Ubiquitin-like Modifier (SUMO dramatically potentiates its ability to transcriptionally repress cyclin D1 and mediate G(1 cell cycle arrest in fibroblast cells. Mutation of SUMO acceptor lysines 159 and 279 located in the C-terminal repression domain has no impact on nuclear localization; however, it abrogates association with the co-repressor histone deacetylase 1 (HDAC1, attenuates repression of cyclin D1, and prevents Stra13-mediated growth suppression. HDAC1, which promotes cellular proliferation and cell cycle progression, antagonizes Stra13 sumoylation-dependent growth arrest. Our results uncover an unidentified regulatory axis between Stra13 and HDAC1 in progression through the G(1/S phase of the cell cycle, and provide new mechanistic insights into regulation of Stra13-mediated transcriptional repression by sumoylation.

  1. KAP1 Is a Host Restriction Factor That Promotes Human Adenovirus E1B-55K SUMO Modification

    DEFF Research Database (Denmark)

    Bürck, Carolin; Mund, Andreas; Berscheminski, Julia

    2016-01-01

    characterized, but represent a decisive moment in establishing a productive infection. Here, we identify a novel host viral restriction factor, KAP1. This heterochromatin associated transcription factor regulates the dynamic organization of host chromatin structure via its ability to influence epigenetic marks...... epigenetic gene silencing and to promote SUMO modification of E1B-55K by a so far unknown mechanism. IMPORTANCE: Here we describe a novel cellular restriction factor for Human Adenovirus (HAdV) that sheds light on very early modulation processes in viral infection. We reported that chromatin formation...... and cellular SWI/SNF chromatin remodeling play a key role in HAdV transcriptional regulation (1-4). We observed that the cellular chromatin-associated factor, and epigenetic reader SPOC1 represses HAdV infection and gene expression. Here, we illustrate the role of the SPOC1 interacting factor KAP1 during...

  2. Senp1 Is Essential for Desumoylating Sumo1-Modified Proteins but Dispensable for Sumo2 and Sumo3 Deconjugation in the Mouse Embryo

    Directory of Open Access Journals (Sweden)

    Prashant Sharma

    2013-05-01

    Full Text Available Posttranslational modification with small ubiquitin-like modifier (Sumo regulates numerous cellular and developmental processes. Sumoylation is dynamic with deconjugation by Sumo-specific proteases (Senps regulating steady-state levels. Different Senps are found in distinct subcellular domains, which may limit their deconjugation activity to colocalizing Sumo-modified proteins. In vitro, Senps can discriminate between the different Sumo paralogs: Sumo1 versus the highly related Sumo2 and Sumo3 (Sumo2/3, which can form poly-Sumo chains. However, a full understanding of Senp specificity in vivo is still lacking. Here, using biochemical and genetic approaches, we establish that Senp1 has an essential, nonredundant function to desumoylate Sumo1-modified proteins during mouse embryonic development. Senp1 specificity for Sumo1 conjugates represents an intrinsic function and not simply a product of colocalization. In contrast, Senp1 has only a limited role in Sumo2/3 desumoylation, although it may regulate Sumo1-mediated termination of poly-Sumo2/3 chains.

  3. SUMO Wrestles with Recombination

    Directory of Open Access Journals (Sweden)

    Lumír Krejčí

    2012-07-01

    Full Text Available DNA double-strand breaks (DSBs comprise one of the most toxic DNA lesions, as the failure to repair a single DSB has detrimental consequences on the cell. Homologous recombination (HR constitutes an error-free repair pathway for the repair of DSBs. On the other hand, when uncontrolled, HR can lead to genome rearrangements and needs to be tightly regulated. In recent years, several proteins involved in different steps of HR have been shown to undergo modification by small ubiquitin-like modifier (SUMO peptide and it has been suggested that deficient sumoylation impairs the progression of HR. This review addresses specific effects of sumoylation on the properties of various HR proteins and describes its importance for the homeostasis of DNA repetitive sequences. The article further illustrates the role of sumoylation in meiotic recombination and the interplay between SUMO and other post-translational modifications.

  4. SUMO-1 possesses DNA binding activity

    Directory of Open Access Journals (Sweden)

    Wieruszeski Jean-Michel

    2010-05-01

    Full Text Available Abstract Background Conjugation of small ubiquitin-related modifiers (SUMOs is a frequent post-translational modification of proteins. SUMOs can also temporally associate with protein-targets via SUMO binding motifs (SBMs. Protein sumoylation has been identified as an important regulatory mechanism especially in the regulation of transcription and the maintenance of genome stability. The precise molecular mechanisms by which SUMO conjugation and association act are, however, not understood. Findings Using NMR spectroscopy and protein-DNA cross-linking experiments, we demonstrate here that SUMO-1 can specifically interact with dsDNA in a sequence-independent fashion. We also show that SUMO-1 binding to DNA can compete with other protein-DNA interactions at the example of the regulatory domain of Thymine-DNA Glycosylase and, based on these competition studies, estimate the DNA binding constant of SUMO1 in the range 1 mM. Conclusion This finding provides an important insight into how SUMO-1 might exert its activity. SUMO-1 might play a general role in destabilizing DNA bound protein complexes thereby operating in a bottle-opener way of fashion, explaining its pivotal role in regulating the activity of many central transcription and DNA repair complexes.

  5. SUMO wrestling in cell movement

    Institute of Scientific and Technical Information of China (English)

    Miia M Rytinki; Jorma J Palvimo

    2011-01-01

    @@ Small ubiquitin-like modifier(SUMO)proteins 1,2 and 3 can be covalently conjugated to specific lysine residues on target proteins in a process dubbed SUMOylation.This conserved posttranslational modification that was reported for the first time in 1996 has emerged as an important regulatory mechanism in cell physiology,especially in nuclear signaling,transport,transcription and DNA replication/repair[1,2].

  6. ATPase-Dependent Control of the Mms21 SUMO Ligase during DNA Repair

    NARCIS (Netherlands)

    M. Bermúdez-López (Marcelino); I. Pociño-Merino (Irene); H. Sanchez (Humberto); A. Bueno (Andrés); C. Guasch (Clàudia); S. Almedawar (Seba); S. Bru-Virgili (Sergi); E. Garí (Eloi); C. Wyman (Claire); D. Reverter (David); N. Colomina (Neus); J. Torres-Rosell (Jordi)

    2015-01-01

    textabstractModification of proteins by SUMO is essential for the maintenance of genome integrity. During DNA replication, the Mms21-branch of the SUMO pathway counteracts recombination intermediates at damaged replication forks, thus facilitating sister chromatid disjunction. The Mms21 SUMO ligase

  7. Interleukin-32α downregulates the activity of the B-cell CLL/lymphoma 6 protein by inhibiting protein kinase Cε-dependent SUMO-2 modification.

    Science.gov (United States)

    Park, Yun Sun; Kang, Jeong-Woo; Lee, Dong Hun; Kim, Man Sub; Bak, Yesol; Yang, Young; Lee, Hee Gu; Hong, JinTae; Yoon, Do-Young

    2014-09-30

    A proinflammatory cytokine IL-32 acts as an intracellular mediator. IL-32α interacts with many intracellular molecules, but there are no reports of interaction with a transcriptional repressor BCL6. In this study, we showed that PMA induces an interaction between IL-32α, PKCε, and BCL6, forming a trimer. To identify the mechanism of the interaction, we treated cells with various inhibitors. In HEK293 and THP-1 cell lines, treatment with a pan-PKC inhibitor, PKCε inhibitor, and PKCδ inhibitor decreased BCL6 and IL-32α protein expression. MAPK inhibitors and classical PKC inhibitor did not decrease PMA-induced BCL6 and IL-32α protein expression. Further, the pan-PKC inhibitor and PKCε inhibitor disrupted PMA-induced interaction between IL-32α and BCL6. These data demonstrate that the intracellular interaction between IL-32α and BCL6 is induced by PMA-activated PKCε. PMA induces post-translational modification of BCL6 by conjugation to SUMO-2, while IL-32α inhibits. PKCε inhibition eliminated PMA-induced SUMOylation of BCL6. Inhibition of BCL6 SUMOylation by IL-32α affected the cellular function and activity of the transcriptional repressor BCL6 in THP-1 cells. Thus, we showed that IL-32α is a negative regulator of the transcriptional repressor BCL6. IL-32α inhibits BCL6 SUMOylation by activating PKCε, resulting in the modulation of BCL6 target genes and cellular functions of BCL6.

  8. Advances on SUMO substrates in Arabidopsis%拟南芥SUMO 底物的研究进展

    Institute of Scientific and Technical Information of China (English)

    郭敏霞; 傅永福

    2013-01-01

    SUMO(Small ubiquitin-related modifier)化修饰是普遍存在于真核生物中的一种翻译后修饰,在很多细胞过程中发挥重要作用.文章阐述了拟南芥中SUMO 底物的最新研究进展.首先介绍SUMO 化修饰过程,SUMO底物的鉴定,包括鉴定方法的进展和鉴定成果.然后依据底物的亚细胞定位、生理功能和参与的生理生化过程对其进行分类和分析,以期更好地理解蛋白质SUMO 化修饰方式在植物生长发育中的功能.%SUMO (Small ubiquitin-related modifier) modification, one of the essential posttranslational modifications in eukaryotes, plays an important role in various cellular processes. This review summarized the recent progresses on SUMO substrates in Arabidopsis. We firstly described the pathway of SUMO conjugation, and then focused on screening for SUMO substrates, including the methods of identification and SUMO substrates identified. Finally, we classified these substrates on the basis of their subcellular localization, functions, and biological processes. It is expected to provide the basis for better understanding the roles of sumoylation of proteins in plants.

  9. The SUMO Pathway in Mitosis.

    Science.gov (United States)

    Mukhopadhyay, Debaditya; Dasso, Mary

    2017-01-01

    Mitosis is the stage of the cell cycle during which replicated chromosomes must be precisely divided to allow the formation of two daughter cells possessing equal genetic material. Much of the careful spatial and temporal organization of mitosis is maintained through post-translational modifications, such as phosphorylation and ubiquitination, of key cellular proteins. Here, we will review evidence that sumoylation, conjugation to the SUMO family of small ubiquitin-like modifiers, also serves essential regulatory roles during mitosis. We will discuss the basic biology of sumoylation, how the SUMO pathway has been implicated in particular mitotic functions, including chromosome condensation, centromere/kinetochore organization and cytokinesis, and what cellular proteins may be the targets underlying these phenomena.

  10. The Ubiquitin-Like SUMO System and Heart Function: From Development to Disease.

    Science.gov (United States)

    Mendler, Luca; Braun, Thomas; Müller, Stefan

    2016-01-01

    SUMOylation is a ubiquitin-related transient posttranslational modification pathway catalyzing the conjugation of small ubiquitin-like modifier (SUMO) proteins (SUMO1, SUMO2, and SUMO3) to lysine residues of proteins. SUMOylation targets a wide variety of cellular regulators and thereby affects a multitude of different cellular processes. SUMO/sentrin-specific proteases are able to remove SUMOs from targets, contributing to a tight control of SUMOylated proteins. Genetic and cell biological experiments indicate a critical role of balanced SUMOylation/deSUMOylation for proper cardiac development, metabolism, and stress adaptation. Here, we review the current knowledge about SUMOylation/deSUMOylation in the heart and provide an integrated picture of cardiac functions of the SUMO system under physiologic or pathologic conditions. We also describe potential therapeutic approaches targeting the SUMO machinery to combat heart disease.

  11. Construction, Expression and Purification of SUMO1-GST Fusion Protein

    Institute of Scientific and Technical Information of China (English)

    QIAO Xiao-fang; FANG Xue-dong; LIU Jun

    2011-01-01

    Sumoylation is an important protein modification discovered recently. SUMO(small ubiquitin-related modifier) pathway regulates the protein stability and transcriptional activity with a 12-kDa small molecular protein,SUMO, ligated to the target protein. The purification of SUMO proteins is a key step to reveal their function. The purpose of this study was to construct the recombinant SUMO1 gene cloned to a pGEX-4T-1 vector to express and purify the SUMO1-GST fusion protein in Escherichia coli. First, the full length DNA sequence of SUMO1 gene was amplified by PCR and was ligated to pMD18-T vector. Then the SUMO1 gene was subcloned to pGEX-4T-1 prokaryotic expression vector between BamHI and XhoI sites, and transformed in Escherichia coli DH5a cells. The right colonies were identified by restrictive enzyme digestion and sequencing. The correct rebombinant plasmid of pGEX-4T-1-SUMO1 was transformed in Escherichia coli BL21 cells and then induced by IPTG(isopropyl-β-D-lthiogalacto-pyranoside) to express the SUMO1-GST fusion protein. The highly purified SUMOl-GST(glutathione S-transferase) fusion protein was obtained by affinity chromatography. Finally, the properties of SUMO1-GST fusion protein were confirmed by Coomassie brilliant blue strain and Western blot analysis. The recombinant plasmid of pGEX-4T-1-SUMO 1 was successfully constructed, and SUMO1-GST fusion proteins were successfully expressed.

  12. USP7 is a SUMO deubiquitinase essential for DNA replication

    DEFF Research Database (Denmark)

    Lecona, Emilio; Rodriguez-Acebes, Sara; Specks, Julia

    2016-01-01

    Post-translational modification of proteins by ubiquitin (Ub) and Ub-like modifiers regulates DNA replication. We have previously shown that chromatin around replisomes is rich in SUMO and poor in Ub, whereas mature chromatin exhibits an opposite pattern. How this SUMO-rich, Ub-poor environment...... is maintained at sites of DNA replication in mammalian cells remains unexplored. Here we identify USP7 as a replisome-enriched SUMO deubiquitinase that is essential for DNA replication. By acting on SUMO and SUMOylated proteins, USP7 counteracts their ubiquitination. Inhibition or genetic deletion of USP7 leads...... to the accumulation of Ub on SUMOylated proteins, which are displaced away from replisomes. Our findings provide a model explaining the differential accumulation of SUMO and Ub at replication forks and identify an essential role of USP7 in DNA replication that should be considered in the development of USP7...

  13. USP7 is a SUMO deubiquitinase essential for DNA replication.

    Science.gov (United States)

    Lecona, Emilio; Rodriguez-Acebes, Sara; Specks, Julia; Lopez-Contreras, Andres J; Ruppen, Isabel; Murga, Matilde; Muñoz, Javier; Mendez, Juan; Fernandez-Capetillo, Oscar

    2016-04-01

    Post-translational modification of proteins by ubiquitin (Ub) and Ub-like modifiers regulates DNA replication. We have previously shown that chromatin around replisomes is rich in SUMO and poor in Ub, whereas mature chromatin exhibits an opposite pattern. How this SUMO-rich, Ub-poor environment is maintained at sites of DNA replication in mammalian cells remains unexplored. Here we identify USP7 as a replisome-enriched SUMO deubiquitinase that is essential for DNA replication. By acting on SUMO and SUMOylated proteins, USP7 counteracts their ubiquitination. Inhibition or genetic deletion of USP7 leads to the accumulation of Ub on SUMOylated proteins, which are displaced away from replisomes. Our findings provide a model explaining the differential accumulation of SUMO and Ub at replication forks and identify an essential role of USP7 in DNA replication that should be considered in the development of USP7 inhibitors as anticancer agents.

  14. Identification and developmental expression of Xenopus laevis SUMO proteases.

    Directory of Open Access Journals (Sweden)

    Yonggang Wang

    Full Text Available SUMO proteins are small ubiquitin-related modifiers. All SUMOs are synthesized as propeptides that are post-translationally cleaved prior to conjugation. After processing, SUMOs become covalently conjugated to cellular targets through a pathway that is similar to ubiquitination. Ubiquitin like protein proteases/Sentrin specific proteases (Ulp/SENPs mediate both processing and deconjugation of SUMOs. The action of Ulp/SENPs makes SUMOylation a highly dynamic post-translational modification. To investigate how Ulp/SENPs are regulated in a developmental context, we isolated and characterized all Ulp/SENPs in Xenopus laevis. Xenopus possess homologues of mammalian SENP3, 5, 6 and 7. All of these enzymes reacted with HA-tagged vinyl sulfone derivatives of SUMO-2 (HA-SU2-VS but not SUMO-1 (HA-SU1-VS, suggesting that they act primarily on SUMO-2 and -3. In contrast, Xenopus possess a single member of the SENP1/SENP2 subfamily of Ulp/SENPs, most closely related to mammalian SENP1. Xenopus SENP1 reacted with HA-SU1-VS and HA-SU2-VS, suggesting that it acts on all SUMO paralogues. We analyzed the mRNA and protein levels for each of the Ulp/SENPs through development; we found that they show distinct patterns of expression that may involve both transcriptional and post-transcriptional regulation. Finally, we have characterized the developmental function of the most abundant Ulp/SENP found within Xenopus eggs, SENP3. Depletion of SENP3 using morpholino antisense oligonucleotides (morpholinos caused accumulation of high molecular weight SUMO-2/3 conjugated species, defects in developing embryos and changes in the expression of some genes regulated by the transforming growth factor beta (TGF-beta pathway. These findings collectively indicate that SUMO proteases are both highly regulated and essential for normal development.

  15. SUMO-Enriched Proteome for Drosophila Innate Immune Response

    Science.gov (United States)

    Handu, Mithila; Kaduskar, Bhagyashree; Ravindranathan, Ramya; Soory, Amarendranath; Giri, Ritika; Elango, Vijay Barathi; Gowda, Harsha; Ratnaparkhi, Girish S.

    2015-01-01

    Small ubiquitin-like modifier (SUMO) modification modulates the expression of defense genes in Drosophila, activated by the Toll/nuclear factor-κB and immune-deficient/nuclear factor-κB signaling networks. We have, however, limited understanding of the SUMO-modulated regulation of the immune response and lack information on SUMO targets in the immune system. In this study, we measured the changes to the SUMO proteome in S2 cells in response to a lipopolysaccharide challenge and identified 1619 unique proteins in SUMO-enriched lysates. A confident set of 710 proteins represents the immune-induced SUMO proteome and analysis suggests that specific protein domains, cellular pathways, and protein complexes respond to immune stress. A small subset of the confident set was validated by in-bacto SUMOylation and shown to be bona-fide SUMO targets. These include components of immune signaling pathways such as Caspar, Jra, Kay, cdc42, p38b, 14-3-3ε, as well as cellular proteins with diverse functions, many being components of protein complexes, such as prosß4, Rps10b, SmD3, Tango7, and Aats-arg. Caspar, a human FAF1 ortholog that negatively regulates immune-deficient signaling, is SUMOylated at K551 and responds to treatment with lipopolysaccharide in cultured cells. Our study is one of the first to describe SUMO proteome for the Drosophila immune response. Our data and analysis provide a global framework for the understanding of SUMO modification in the host response to pathogens. PMID:26290570

  16. SUMO-2 and PIAS1 modulate insoluble mutant huntingtin protein accumulation.

    Science.gov (United States)

    O'Rourke, Jacqueline Gire; Gareau, Jaclyn R; Ochaba, Joseph; Song, Wan; Raskó, Tamás; Reverter, David; Lee, John; Monteys, Alex Mas; Pallos, Judit; Mee, Lisa; Vashishtha, Malini; Apostol, Barbara L; Nicholson, Thomas Peter; Illes, Katalin; Zhu, Ya-Zhen; Dasso, Mary; Bates, Gillian P; Difiglia, Marian; Davidson, Beverly; Wanker, Erich E; Marsh, J Lawrence; Lima, Christopher D; Steffan, Joan S; Thompson, Leslie M

    2013-07-25

    A key feature in Huntington disease (HD) is the accumulation of mutant Huntingtin (HTT) protein, which may be regulated by posttranslational modifications. Here, we define the primary sites of SUMO modification in the amino-terminal domain of HTT, show modification downstream of this domain, and demonstrate that HTT is modified by the stress-inducible SUMO-2. A systematic study of E3 SUMO ligases demonstrates that PIAS1 is an E3 SUMO ligase for both HTT SUMO-1 and SUMO-2 modification and that reduction of dPIAS in a mutant HTT Drosophila model is protective. SUMO-2 modification regulates accumulation of insoluble HTT in HeLa cells in a manner that mimics proteasome inhibition and can be modulated by overexpression and acute knockdown of PIAS1. Finally, the accumulation of SUMO-2-modified proteins in the insoluble fraction of HD postmortem striata implicates SUMO-2 modification in the age-related pathogenic accumulation of mutant HTT and other cellular proteins that occurs during HD progression.

  17. SUMO-2 and PIAS1 Modulate Insoluble Mutant Huntingtin Protein Accumulation

    Directory of Open Access Journals (Sweden)

    Jacqueline Gire O’Rourke

    2013-07-01

    Full Text Available A key feature in Huntington disease (HD is the accumulation of mutant Huntingtin (HTT protein, which may be regulated by posttranslational modifications. Here, we define the primary sites of SUMO modification in the amino-terminal domain of HTT, show modification downstream of this domain, and demonstrate that HTT is modified by the stress-inducible SUMO-2. A systematic study of E3 SUMO ligases demonstrates that PIAS1 is an E3 SUMO ligase for both HTT SUMO-1 and SUMO-2 modification and that reduction of dPIAS in a mutant HTT Drosophila model is protective. SUMO-2 modification regulates accumulation of insoluble HTT in HeLa cells in a manner that mimics proteasome inhibition and can be modulated by overexpression and acute knockdown of PIAS1. Finally, the accumulation of SUMO-2-modified proteins in the insoluble fraction of HD postmortem striata implicates SUMO-2 modification in the age-related pathogenic accumulation of mutant HTT and other cellular proteins that occurs during HD progression.

  18. SUMO-2 Orchestrates Chromatin Modifiers in Response to DNA Damage

    DEFF Research Database (Denmark)

    Hendriks, Ivo A; Treffers, Louise W; Verlaan-de Vries, Matty

    2015-01-01

    with the DNA damaging agent methyl methanesulfonate (MMS). We have uncovered a dynamic set of 20 upregulated and 33 downregulated SUMO-2 conjugates, and 755 SUMO-2 sites, of which 362 were dynamic in response to MMS. In contrast to yeast, where a response is centered on homologous recombination, we identified......Small ubiquitin-like modifiers play critical roles in the DNA damage response (DDR). To increase our understanding of SUMOylation in the mammalian DDR, we employed a quantitative proteomics approach in order to identify dynamically regulated SUMO-2 conjugates and modification sites upon treatment...... dynamically SUMOylated interaction networks of chromatin modifiers, transcription factors, DNA repair factors, and nuclear body components. SUMOylated chromatin modifiers include JARID1B/KDM5B, JARID1C/KDM5C, p300, CBP, PARP1, SetDB1, and MBD1. Whereas SUMOylated JARID1B was ubiquitylated by the SUMO...

  19. High Glucose Induces Sumoylation of Smad4 via SUMO2/3 in Mesangial Cells

    Directory of Open Access Journals (Sweden)

    Xueqin Zhou

    2014-01-01

    Full Text Available Recent studies have shown that sumoylation is a posttranslational modification involved in regulation of the transforming growth factor-β (TGF-β signaling pathway, which plays a critical role in renal fibrosis in diabetic nephropathy (DN. However, the role of sumoylation in the regulation of TGF-β signaling in DN is still unclear. In the present study, we investigated the expression of SUMO (SUMO1 and SUMO2/3 and Smad4 and the interaction between SUMO and Smad4 in cultured rat mesangial cells induced by high glucose. We found that SUMO1 and SUMO2/3 expression was significantly increased in the high glucose groups compared to the normal group P<0.05. Smad4 and fibronectin (FN levels were also increased in the high glucose groups in a dose-dependent manner. Coimmunoprecipitation and confocal laser scanning revealed that Smad4 interacted and colocalized with SUMO2/3, but not with SUMO1 in mesangial cells. Sumoylation (SUMO2/3 of Smad4 under high glucose condition was strongly enhanced compared to normal control P<0.05. These results suggest that high glucose may activate TGF-β/Smad signaling through sumoylation of Samd4 by SUMO2/3 in mesangial cells.

  20. Construction of a mouse Aos1-Uba2 chimeric SUMO-E1 enzyme, mAU, and its expression in baculovirus-insect cells

    Science.gov (United States)

    Nakayama, Tomofumi; Yuasa, Eri; Kanemaru, Ayumi; Saito, Masayuki; Saitoh, Hisato

    2014-01-01

    Small ubiquitin-related modifier (SUMO) is a highly conserved protein that is covalently attached to target proteins. This posttranslational modification, designated SUMOylation, is a major protein-conjugation-driven strategy designed to regulate structure and function of cellular proteins. SUMOylation consists of an enzymatic cascade involving the E1-activating enzyme and the E2-conjugating enzyme. The SUMO-E1 enzyme consists of two subunits, a heterodimer of activation of Smt3p 1 (Aos1) and ubiquitin activating enzyme 2 (Uba2), which resembles the N- and C-terminal halves of ubiquitin E1 (Uba1). Herein, we describe the rational design of a single polypeptide version of SUMO-E1, a chimera of mouse Aos1 and Uba2 subunits, termed mAU, in which the functional domains appear to be arranged in a fashion similar to Uba1. We also describe the construction of a mAU plasmid for expression in a baculovirus-insect cell system and present an in situ SUMOylation assay using the recombinant mAU. Our results showed that mAU has SUMO-E1 activity, thereby indicating that mAU can be expressed in baculovirus-insect cells and represents a suitable source of SUMO-E1. PMID:24637489

  1. Meiosis and SUMO

    DEFF Research Database (Denmark)

    Holm, Lærke Rebekka

    to target proteins can be catalyzed by the SUMO E3 ligase Pli1. In this study we investigate the role of Pli1 and Pmt3 during meiotic differentiation and at repetitive DNA during mitotic growth. Target proteins for Pmt3 are many; however, Pli1 has a meiosis-specic function regulating meiotic recombination...

  2. Meiosis and SUMO

    DEFF Research Database (Denmark)

    Holm, Lærke Rebekka

    to target proteins can be catalyzed by the SUMO E3 ligase Pli1. In this study we investigate the role of Pli1 and Pmt3 during meiotic differentiation and at repetitive DNA during mitotic growth. Target proteins for Pmt3 are many; however, Pli1 has a meiosis-specic function regulating meiotic recombination...

  3. Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells

    DEFF Research Database (Denmark)

    Bursomanno, Sara; Beli, Petra; Khan, Asif M;

    2015-01-01

    SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related...... subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic...... repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability...

  4. 2nd SUMO Conference

    CERN Document Server

    Weber, Melanie

    2015-01-01

    This contributed volume contains the conference proceedings of the Simulation of Urban Mobility (SUMO) conference 2014, Berlin. The included research papers cover a wide range of topics in traffic planning and simulation, including open data, vehicular communication, e-mobility, urban mobility, multimodal traffic as well as usage approaches. The target audience primarily comprises researchers and experts in the field, but the book may also be beneficial for graduate students.  

  5. Comprehensive Identification of SUMO2/3 Targets and Their Dynamics during Mitosis

    DEFF Research Database (Denmark)

    Schou, Julie; Kelstrup, Christian D; Hayward, Daniel G

    2014-01-01

    During mitosis large alterations in cellular structures occur rapidly, which to a large extent is regulated by post-translational modification of proteins. Modification of proteins with the small ubiquitin-related protein SUMO2/3 regulates mitotic progression, but few mitotic targets have been...... identified so far. To deepen our understanding of SUMO2/3 during this window of the cell cycle, we undertook a comprehensive proteomic characterization of SUMO2/3 modified proteins in mitosis and upon mitotic exit. We developed an efficient tandem affinity purification strategy of SUMO2/3 modified proteins...... from mitotic cells. Combining this purification strategy with cell synchronization procedures and quantitative mass spectrometry allowed for the mapping of numerous novel targets and their dynamics as cells progressed out of mitosis. This identified RhoGDIα as a major SUMO2/3 modified protein...

  6. A monoclonal antibody against the human SUMO-1 protein obtained by immunization with recombinant protein and CpG-DNA-liposome complex.

    Science.gov (United States)

    Kim, Dongbum; Lee, Joo Young; Song, Dae-Geun; Kwon, Sanghoon; Lee, Younghee; Pan, Cheol-Ho; Kwon, Hyung-Joo

    2013-10-01

    Post-translational modification regulated by conjugation of a small ubiquitin-like modifier (SUMO) is involved in various cellular processes. In this study, we expressed and purified recombinant human SUMO-1 (hSUMO-1). BALB/c mice were immunized with a complex of hSUMO-1 protein and Lipoplex(O) to produce hSUMO-1-specific antibodies. Using conventional hybridoma technology, we obtained four hybridoma clones derived from the mouse with the highest antibody titer against hSUMO-1. Based on Western blot analysis, our hSUMO-1 monoclonal antibody specifically recognizes hSUMO-1, but not other SUMO proteins. These results support that the anti-hSUMO-1 monoclonal antibody produced with the aid of Lipoplex(O) adjuvant is specific and that Lipoplex(O) is useful for development of monoclonal antibodies against recombinant protein. In addition, we analyzed human tissues to examine the distribution of hSUMO-1. Higher expression of hSUMO-1 was detected in normal adrenal gland, esophagus, pancreas, liver, stomach, kidney, and uterus than in corresponding cancer tissues, suggesting a tumor suppressive function of hSUMO-1.

  7. 丙烯醛调控糖皮质激素受体类小泛素化水平对气道黏液高分泌的影响%Acrolein decreases SUMO modification of glucocorticoid receptor and reduces its sensitivity to corticosteroids in airway hypersecretion

    Institute of Scientific and Technical Information of China (English)

    许瑞; 李琪; 周向东; 尤列·皮尔曼; 维克多·科罗索夫

    2012-01-01

    Objective To explore the variation of small ubiquitin-related modification (SUMO) level of glucocorticoid receptors (GRs) exposed to acrolein stimulation as well as the influence of the variation on mucus hypersecretion.Methods The recombinant plasmid was constructed by inserting small ubiquitin-like modifier 1 (SUMO1) cDNA into eukaryotic expression plasmid pcDNA3.1-EGFP with a flag sequence marker.The recombinant plasmid pcDNA3.1-EGFP-flag-SUMO1 was identified by enzyme digestion analysis.The 16HBE cells were cultured and randomized divided into 9 following groups:A.control; B.acrolein stimulated; C.dexamethasone incubated; D.acrolein stimulated and dexamethasone incubated; E.SUMO1 transfected; F.pcDNA3.1-EGFP vector transfected; G.SUMO1 transfected and acrolein stimulated; H.SUMO1 transfected,acrolein stimulated and dexamethasone incubated; Ⅰ.pcDNA3.1-EGFP vector transfected,acrolein stimulated and dexamethasone incubated.Each group consisted of 5 parallel wells and experiments were repeated for 4 times for statistical analysis.The transfection efficiency was evaluated by the expression of enhanced green fluorescent protein (EGFP) via fluorescence microscope.The SUMO modification level of GRs was measured with co-immunoprecipitation and Western blot.The transcription level of mucin (MUC) 5AC was evaluated with reverse transcriptionpolymerase chain reaction (RT-PCR).The MUC5AC secreted in supernatant was measured by enzymelinked immunosorbent assay (ELISA).Results The SUMO modification level was much lower in group B (0.079 ± 0.023) than that in group A (0.446 ± 0.068) (t =-23.13,P =0.000) and in group D (0.574 ±0.018) than that in group C (0.843 ±0.028) (t =-36.34,P =0.000).The transcription level of MUC5AC was significantly lower in group H (0.330 ±0.063) than group D (0.617 ± 0.190) (q =-7.80,P =0.000).Through ELISA,there was no significance between groups G and B in the term of secretion level of MUC5AC.While the secretion level of MUC5AC was much

  8. Comprehensive identification of SUMO2/3 targets and their dynamics during mitosis.

    Science.gov (United States)

    Schou, Julie; Kelstrup, Christian D; Hayward, Daniel G; Olsen, Jesper V; Nilsson, Jakob

    2014-01-01

    During mitosis large alterations in cellular structures occur rapidly, which to a large extent is regulated by post-translational modification of proteins. Modification of proteins with the small ubiquitin-related protein SUMO2/3 regulates mitotic progression, but few mitotic targets have been identified so far. To deepen our understanding of SUMO2/3 during this window of the cell cycle, we undertook a comprehensive proteomic characterization of SUMO2/3 modified proteins in mitosis and upon mitotic exit. We developed an efficient tandem affinity purification strategy of SUMO2/3 modified proteins from mitotic cells. Combining this purification strategy with cell synchronization procedures and quantitative mass spectrometry allowed for the mapping of numerous novel targets and their dynamics as cells progressed out of mitosis. This identified RhoGDIα as a major SUMO2/3 modified protein, specifically during mitosis, mediated by the SUMO ligases PIAS2 and PIAS3. Our data provide a rich resource for further exploring the role of SUMO2/3 modifications in mitosis and cell cycle regulation.

  9. SUMO fusion technology for enhanced protein production in prokaryotic and eukaryotic expression systems.

    Science.gov (United States)

    Panavas, Tadas; Sanders, Carsten; Butt, Tauseef R

    2009-01-01

    In eukaryotic cells, the reversible attachment of small ubiquitin-like modifier (SUMO) protein is a post-translational modification that has been demonstrated to play an important role in various cellular processes. Moreover, it has been found that SUMO as an N-terminal fusion partner enhances functional protein production in prokaryotic and eukaryotic expression systems, based upon significantly improved protein stability and solubility. Following the expression and purification of the fusion protein, the SUMO-tag can be cleaved by specific (SUMO) proteases via their endopeptidase activity in vitro to generate the desired N-terminus of the released protein partner. In addition to its physiological relevance in eukaryotes, SUMO can, thus, be used as a powerful biotechnological tool for protein expression in prokaryotic and eukaryotic cell systems.In this chapter, we will describe the construction of a fusion protein with the SUMO-tag, its expression in Escherichia coli, and its purification followed by the removal of the SUMO-tag by a SUMO-specific protease in vitro.

  10. Joosep ja sumo / Mart Velsker

    Index Scriptorium Estoniae

    Velsker, Mart, 1966-

    2008-01-01

    Arvustus: Kivisildnik. Sumo. Pärnu : Jumalikud Ilmutused, 2007 ; Rilke, Rainer Maria. Joosepi kahtlused / tõlk. Kivisildnik. Pärnu : Jumalikud Ilmutused, 2008. - Artikkel sisaldab bibliograafiat joonealustes viidetes

  11. The potential role of As-sumo-1 in the embryonic diapause process and early embryo development of Artemia sinica.

    Science.gov (United States)

    Chu, Bing; Yao, Feng; Cheng, Cheng; Wu, Yang; Mei, Yanli; Li, Xuejie; Liu, Yan; Wang, Peisheng; Hou, Lin; Zou, Xiangyang

    2014-01-01

    During embryonic development of Artemia sinica, environmental stresses induce the embryo diapause phenomenon, required to resist apoptosis and regulate cell cycle activity. The small ubiquitin-related modifier-1 (SUMO), a reversible post-translational protein modifier, plays an important role in embryo development. SUMO regulates multiple cellular processes, including development and other biological processes. The molecular mechanism of diapause, diapause termination and the role of As-sumo-1 in this processes and in early embryo development of Artemia sinica still remains unknown. In this study, the complete cDNA sequences of the sumo-1 homolog, sumo ligase homolog, caspase-1 homolog and cyclin B homolog from Artemia sinica were cloned. The mRNA expression patterns of As-sumo-1, sumo ligase, caspase-1, cyclin B and the location of As-sumo-1 were investigated. SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E proteins were analyzed during different developmental stages of the embryo of A. sinica. Small interfering RNA (siRNA) was used to verify the function of sumo-1 in A. sinica. The full-length cDNA of As-sumo-1 was 476 bp, encoding a 92 amino acid protein. The As-caspases-1 cDNA was 966 bp, encoding a 245 amino-acid protein. The As-sumo ligase cDNA was 1556 bp encoding, a 343 amino acid protein, and the cyclin B cDNA was 739 bp, encoding a 133 amino acid protein. The expressions of As-sumo-1, As-caspase-1 and As-cyclin B were highest at the 10 h stage of embryonic development, and As-sumo ligase showed its highest expression at 0 h. The expression of As-SUMO-1 showed no tissue or organ specificity. Western blotting showed high expression of As-SUMO-1, p53, Mdm2, Caspase-1, Cyclin B and Cyclin E at the 10 h stage. The siRNA caused abnormal development of the embryo, with increased malformation and mortality. As-SUMO-1 is a crucial regulation and modification protein resumption of embryonic diapause and early embryo development of A. sinica.

  12. At the crossroads of SUMO and NF-κB

    Directory of Open Access Journals (Sweden)

    Schmidt Christian

    2003-11-01

    Full Text Available Abstract Background Recognition of pathogens by immune receptors leads to activation of macrophages, dendritic cells, and lymphocytes. Signals are communicated to enhance expression of target molecules such as cytokines and adhesion molecules, depending on activation of various inducible transcription factors, among which the family NF-κB transcription factors plays an evolutionarily conserved and critical role. Classical activation of NF-κB involves phosphorylation, polyubiquitination and subsequent degradation of the inhibitor molecules of NF-κB, referred to as IκB. Modification of IκBα, one of the mammalian IκB isoforms, with the small ubiquitin-like modifier (SUMO results its protection from degradation. Presentation of the hypothesis SUMO-IκBα localizes in the nucleus. The nuclear SUMO-IκBα pool may be dynamic. SUMO-IκBα functions as synergy control factor. Testing the hypothesis Immunoprecipitation from cellular fractions, 35S methionine pulse-chase, and FRET assays should reveal the localization of SUMO-IκBα and the dynamics of the pool. Expression of SUMOylation defective IκBα in an IκBα -/- background should yield insights into the function of SUMO-IκBα. Implication of the hypothesis IκBα contains the required SUMOylation motif but IκBβ does not. The suggested study would provide evidence whether or not IκBα and IκBβ can substitute each other. In addition, the suggested assays would reveal a possible redundancy in controlling transcriptional activity of NF-κB.

  13. Cell cycle-dependent SUMO-1 conjugation to nuclear mitotic apparatus protein (NuMA)

    Energy Technology Data Exchange (ETDEWEB)

    Seo, Jae Sung; Kim, Ha Na; Kim, Sun-Jick; Bang, Jiyoung; Kim, Eun-A; Sung, Ki Sa [Department of Biological Sciences, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Yoon, Hyun-Joo [TissueGene Inc. 9605 Medical Center Dr., Rockville, MD 20850 (United States); Yoo, Hae Yong [Department of Health Sciences and Technology, Samsung Advanced Institute for Health Sciences and Technology, Samsung Medical Center, Sungkyunkwan University, Seoul 135-710 (Korea, Republic of); Choi, Cheol Yong, E-mail: choicy@skku.ac.kr [Department of Biological Sciences, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of)

    2014-01-03

    Highlights: •NuMA is modified by SUMO-1 in a cell cycle-dependent manner. •NuMA lysine 1766 is the primary target site for SUMOylation. •SUMOylation-deficient NuMA induces multiple spindle poles during mitosis. •SUMOylated NuMA induces microtubule bundling. -- Abstract: Covalent conjugation of proteins with small ubiquitin-like modifier 1 (SUMO-1) plays a critical role in a variety of cellular functions including cell cycle control, replication, and transcriptional regulation. Nuclear mitotic apparatus protein (NuMA) localizes to spindle poles during mitosis, and is an essential component in the formation and maintenance of mitotic spindle poles. Here we show that NuMA is a target for covalent conjugation to SUMO-1. We find that the lysine 1766 residue is the primary NuMA acceptor site for SUMO-1 conjugation. Interestingly, SUMO modification of endogenous NuMA occurs at the entry into mitosis and this modification is reversed after exiting from mitosis. Knockdown of Ubc9 or forced expression of SENP1 results in impairment of the localization of NuMA to mitotic spindle poles during mitosis. The SUMOylation-deficient NuMA mutant is defective in microtubule bundling, and multiple spindles are induced during mitosis. The mitosis-dependent dynamic SUMO-1 modification of NuMA might contribute to NuMA-mediated formation and maintenance of mitotic spindle poles during mitosis.

  14. Quantitative Förster resonance energy transfer analysis for kinetic determinations of SUMO-specific protease.

    Science.gov (United States)

    Liu, Yan; Song, Yang; Madahar, Vipul; Liao, Jiayu

    2012-03-01

    Förster resonance energy transfer (FRET) technology has been widely used in biological and biomedical research, and it is a very powerful tool for elucidating protein interactions in either dynamic or steady state. SUMOylation (the process of SUMO [small ubiquitin-like modifier] conjugation to substrates) is an important posttranslational protein modification with critical roles in multiple biological processes. Conjugating SUMO to substrates requires an enzymatic cascade. Sentrin/SUMO-specific proteases (SENPs) act as an endopeptidase to process the pre-SUMO or as an isopeptidase to deconjugate SUMO from its substrate. To fully understand the roles of SENPs in the SUMOylation cycle, it is critical to understand their kinetics. Here, we report a novel development of a quantitative FRET-based protease assay for SENP1 kinetic parameter determination. The assay is based on the quantitative analysis of the FRET signal from the total fluorescent signal at acceptor emission wavelength, which consists of three components: donor (CyPet-SUMO1) emission, acceptor (YPet) emission, and FRET signal during the digestion process. Subsequently, we developed novel theoretical and experimental procedures to determine the kinetic parameters, k(cat), K(M), and catalytic efficiency (k(cat)/K(M)) of catalytic domain SENP1 toward pre-SUMO1. Importantly, the general principles of this quantitative FRET-based protease kinetic determination can be applied to other proteases. Copyright © 2011 Elsevier Inc. All rights reserved.

  15. Cerebral ischemia/stroke and small ubiquitin-like modifier (SUMO) conjugation--a new target for therapeutic intervention?

    Science.gov (United States)

    Yang, Wei; Sheng, Huaxin; Homi, H Mayumi; Warner, David S; Paschen, Wulf

    2008-08-01

    Transient cerebral ischemia/stroke activates various post-translational protein modifications such as phosphorylation and ubiquitin conjugation that are believed to play a major role in the pathological process triggered by an interruption of blood supply and culminating in cell death. A new system of post-translational protein modification has been identified, termed as small ubiquitin-like modifier (SUMO) conjugation. Like ubiquitin, SUMO is conjugated to the lysine residue of target proteins in a complex process. This review summarizes observations from recent experiments focusing on the effect of cerebral ischemia on SUMO conjugation. Transient global and focal cerebral ischemia both induced a rapid, dramatic and long-lasting rise in levels of SUMO2/3 conjugation. After transient focal cerebral ischemia, SUMO conjugation was particularly prominent in neurons located at the border of the ischemic territory where SUMO-conjugated proteins translocated to the nucleus. Many SUMO conjugation target proteins are transcription factors and sumoylation has been shown to have a major impact on the activity, stability, and cellular localization of target proteins. The rise in levels of SUMO-conjugated proteins is therefore likely to have a major effect on the fate of post-ischemic neurons. The sumoylation process could provide an exciting new target for therapeutic intervention.

  16. Viral Mimicry to Usurp Ubiquitin and SUMO Host Pathways

    Directory of Open Access Journals (Sweden)

    Peter Wimmer

    2015-08-01

    Full Text Available Posttranslational modifications (PTMs of proteins include enzymatic changes by covalent addition of cellular regulatory determinants such as ubiquitin (Ub and small ubiquitin-like modifier (SUMO moieties. These modifications are widely used by eukaryotic cells to control the functional repertoire of proteins. Over the last decade, it became apparent that the repertoire of ubiquitiylation and SUMOylation regulating various biological functions is not restricted to eukaryotic cells, but is also a feature of human virus families, used to extensively exploit complex host-cell networks and homeostasis. Intriguingly, besides binding to host SUMO/Ub control proteins and interfering with the respective enzymatic cascade, many viral proteins mimic key regulatory factors to usurp this host machinery and promote efficient viral outcomes. Advanced detection methods and functional studies of ubiquitiylation and SUMOylation during virus-host interplay have revealed that human viruses have evolved a large arsenal of strategies to exploit these specific PTM processes. In this review, we highlight the known viral analogs orchestrating ubiquitin and SUMO conjugation events to subvert and utilize basic enzymatic pathways.

  17. Proteome-wide analysis of SUMO2 targets in response to pathological DNA replication stress in human cells.

    Science.gov (United States)

    Bursomanno, Sara; Beli, Petra; Khan, Asif M; Minocherhomji, Sheroy; Wagner, Sebastian A; Bekker-Jensen, Simon; Mailand, Niels; Choudhary, Chunaram; Hickson, Ian D; Liu, Ying

    2015-01-01

    SUMOylation is a form of post-translational modification involving covalent attachment of SUMO (Small Ubiquitin-like Modifier) polypeptides to specific lysine residues in the target protein. In human cells, there are four SUMO proteins, SUMO1-4, with SUMO2 and SUMO3 forming a closely related subfamily. SUMO2/3, in contrast to SUMO1, are predominantly involved in the cellular response to certain stresses, including heat shock. Substantial evidence from studies in yeast has shown that SUMOylation plays an important role in the regulation of DNA replication and repair. Here, we report a proteomic analysis of proteins modified by SUMO2 in response to DNA replication stress in S phase in human cells. We have identified a panel of 22 SUMO2 targets with increased SUMOylation during DNA replication stress, many of which play key functions within the DNA replication machinery and/or in the cellular response to DNA damage. Interestingly, POLD3 was found modified most significantly in response to a low dose aphidicolin treatment protocol that promotes common fragile site (CFS) breakage. POLD3 is the human ortholog of POL32 in budding yeast, and has been shown to act during break-induced recombinational repair. We have also shown that deficiency of POLD3 leads to an increase in RPA-bound ssDNA when cells are under replication stress, suggesting that POLD3 plays a role in the cellular response to DNA replication stress. Considering that DNA replication stress is a source of genome instability, and that excessive replication stress is a hallmark of pre-neoplastic and tumor cells, our characterization of SUMO2 targets during a perturbed S-phase should provide a valuable resource for future functional studies in the fields of DNA metabolism and cancer biology.

  18. SUMO meets meiosis: an encounter at the synaptonemal complex: SUMO chains and sumoylated proteins suggest that heterogeneous and complex interactions lie at the centre of the synaptonemal complex.

    Science.gov (United States)

    Watts, Felicity Z; Hoffmann, Eva

    2011-07-01

    Recent discoveries have identified the small ubiquitin-like modifier (SUMO) as the potential 'missing link' that could explain how the synaptonemal complex (SC) is formed during meiosis. The SC is important for a variety of chromosome interactions during meiosis and appears ladder-like. It is formed when 'axes' of the two homologous chromosomes become connected by the deposition of transverse filaments, forming the steps of the ladder. Although several components of axial and transverse elements have been identified, how the two are connected to form the SC has remained an enigma. Recent discoveries suggest that SUMO modification underlies protein-protein interactions within the SC of budding yeast. The versatility of SUMO in regulating protein-protein interactions adds an exciting new dimension to our understanding of the SC and suggests that SCs are not homogenous structures throughout the nucleus. We propose that this heterogeneity may allow differential regulation of chromosome structure and function.

  19. Identification of SUMO Targets by a Novel Proteomic Approach in Plants

    Institute of Scientific and Technical Information of China (English)

    Gema López-Torrejón; Davide Guerra; Rafael Catalá; Julio Salinas; Juan C.del Pozo

    2013-01-01

    Post-translational modifications (PTMs) chemically and physically alter the properties of proteins,including their folding,subcellular localization,stability,activity,and consequently their function.In spite of their relevance,studies on PTMs in plants are still limited.Small Ubiquitin-like Modifier (SUMO) modification regulates several biological processes by affecting protein-protein interactions,or changing the subcellular localizations of the target proteins.Here,we describe a novel proteomic approach to identify SUMO targets that combines 2-D liquid chromatography,immunodetection,and mass spectrometry (MS) analyses.We have applied this approach to identify nuclear SUMO targets in response to heat shock.Using a bacterial SUMOylation system,we validated that some of the targets identified here are,in fact,labeled with SUMO1.Interestingly,we found that GIGANTEA (GI),a photoperiodic-pathway protein,is modified with SUMO in response to heat shock both in vitro and in vivo.

  20. Defining the SUMO System in Maize: SUMOylation Is Up-Regulated during Endosperm Development and Rapidly Induced by Stress.

    Science.gov (United States)

    Augustine, Robert C; York, Samuel L; Rytz, Thérèse C; Vierstra, Richard D

    2016-07-01

    In response to abiotic and biotic challenges, plants rapidly attach small ubiquitin-related modifier (SUMO) to a large collection of nuclear proteins, with studies in Arabidopsis (Arabidopsis thaliana) linking SUMOylation to stress tolerance via its modification of factors involved in chromatin and RNA dynamics. Despite this importance, little is known about SUMOylation in crop species. Here, we describe the plant SUMO system at the phylogenetic, biochemical, and transcriptional levels with a focus on maize (Zea mays). In addition to canonical SUMOs, land plants encode a loosely constrained noncanonical isoform and a variant containing a long extension upstream of the signature β-grasp fold, with cereals also expressing a novel diSUMO polypeptide bearing two SUMO β-grasp domains in tandem. Maize and other cereals also synthesize a unique SUMO-conjugating enzyme variant with more restricted expression patterns that is enzymatically active despite a distinct electrostatic surface. Maize SUMOylation primarily impacts nuclear substrates, is strongly induced by high temperatures, and displays a memory that suppresses subsequent conjugation. Both in-depth transcript and conjugate profiles in various maize organs point to tissue/cell-specific functions for SUMOylation, with potentially significant roles during embryo and endosperm maturation. Collectively, these studies define the organization of the maize SUMO system and imply important functions during seed development and stress defense.

  1. Identification of SUMO target proteins by quantitative proteomics

    DEFF Research Database (Denmark)

    Andersen, Jens S; Matic, Ivan; Vertegaal, Alfred C O

    2009-01-01

    The identification of target proteins for small ubiquitin-like modifiers (SUMOs) is a critical step towards a detailed understanding of the cellular functions of SUMOs. Substrate protein identification for SUMOs is hampered by the low abundance of SUMO targets, the finding that only a small fract...

  2. GPS-SUMO: a tool for the prediction of sumoylation sites and SUMO-interaction motifs.

    Science.gov (United States)

    Zhao, Qi; Xie, Yubin; Zheng, Yueyuan; Jiang, Shuai; Liu, Wenzhong; Mu, Weiping; Liu, Zexian; Zhao, Yong; Xue, Yu; Ren, Jian

    2014-07-01

    Small ubiquitin-like modifiers (SUMOs) regulate a variety of cellular processes through two distinct mechanisms, including covalent sumoylation and non-covalent SUMO interaction. The complexity of SUMO regulations has greatly hampered the large-scale identification of SUMO substrates or interaction partners on a proteome-wide level. In this work, we developed a new tool called GPS-SUMO for the prediction of both sumoylation sites and SUMO-interaction motifs (SIMs) in proteins. To obtain an accurate performance, a new generation group-based prediction system (GPS) algorithm integrated with Particle Swarm Optimization approach was applied. By critical evaluation and comparison, GPS-SUMO was demonstrated to be substantially superior against other existing tools and methods. With the help of GPS-SUMO, it is now possible to further investigate the relationship between sumoylation and SUMO interaction processes. A web service of GPS-SUMO was implemented in PHP+JavaScript and freely available at http://sumosp.biocuckoo.org.

  3. Characterization of the loss of SUMO pathway function on cancer cells and tumor proliferation.

    Science.gov (United States)

    He, Xingyue; Riceberg, Jessica; Pulukuri, Sai M; Grossman, Steve; Shinde, Vaishali; Shah, Pooja; Brownell, James E; Dick, Larry; Newcomb, John; Bence, Neil

    2015-01-01

    SUMOylation is a post-translational ubiquitin-like protein modification pathway that regulates important cellular processes including chromosome structure, kinetochore function, chromosome segregation, nuclear and sub-nuclear organization, transcription and DNA damage repair. There is increasing evidence that the SUMO pathway is dysregulated in cancer, raising the possibility that modulation of this pathway may have therapeutic potential. To investigate the importance of the SUMO pathway in the context of cancer cell proliferation and tumor growth, we applied lentivirus-based short hairpin RNAs (shRNA) to knockdown SUMO pathway genes in human cancer cells. shRNAs for SAE2 and UBC9 reduced SUMO conjugation activity and inhibited proliferation of human cancer cells. To expand upon these observations, we generated doxycycline inducible conditional shRNA cell lines for SAE2 to achieve acute and reversible SAE2 knockdown. Conditional SAE2 knockdown in U2OS and HCT116 cells slowed cell growth in vitro, and SAE2 knockdown induced multiple terminal outcomes including apoptosis, endoreduplication and senescence. Multinucleated cells became senescent and stained positive for the senescence marker, SA-β Gal, and displayed elevated levels of p53 and p21. In an attempt to explain these phenotypes, we confirmed that loss of SUMO pathway activity leads to a loss of SUMOylated Topoisomerase IIα and the appearance of chromatin bridges which can impair proper cytokinesis and lead to multinucleation. Furthermore, knockdown of SAE2 induces disruption of PML nuclear bodies which may further promote apoptosis or senescence. In an in vivo HCT116 xenograft tumor model, conditional SAE2 knockdown strongly impaired tumor growth. These data demonstrate that the SUMO pathway is required for cancer cell proliferation in vitro and tumor growth in vivo, implicating the SUMO pathway as a potential cancer therapeutic target.

  4. Characterization of the loss of SUMO pathway function on cancer cells and tumor proliferation.

    Directory of Open Access Journals (Sweden)

    Xingyue He

    Full Text Available SUMOylation is a post-translational ubiquitin-like protein modification pathway that regulates important cellular processes including chromosome structure, kinetochore function, chromosome segregation, nuclear and sub-nuclear organization, transcription and DNA damage repair. There is increasing evidence that the SUMO pathway is dysregulated in cancer, raising the possibility that modulation of this pathway may have therapeutic potential. To investigate the importance of the SUMO pathway in the context of cancer cell proliferation and tumor growth, we applied lentivirus-based short hairpin RNAs (shRNA to knockdown SUMO pathway genes in human cancer cells. shRNAs for SAE2 and UBC9 reduced SUMO conjugation activity and inhibited proliferation of human cancer cells. To expand upon these observations, we generated doxycycline inducible conditional shRNA cell lines for SAE2 to achieve acute and reversible SAE2 knockdown. Conditional SAE2 knockdown in U2OS and HCT116 cells slowed cell growth in vitro, and SAE2 knockdown induced multiple terminal outcomes including apoptosis, endoreduplication and senescence. Multinucleated cells became senescent and stained positive for the senescence marker, SA-β Gal, and displayed elevated levels of p53 and p21. In an attempt to explain these phenotypes, we confirmed that loss of SUMO pathway activity leads to a loss of SUMOylated Topoisomerase IIα and the appearance of chromatin bridges which can impair proper cytokinesis and lead to multinucleation. Furthermore, knockdown of SAE2 induces disruption of PML nuclear bodies which may further promote apoptosis or senescence. In an in vivo HCT116 xenograft tumor model, conditional SAE2 knockdown strongly impaired tumor growth. These data demonstrate that the SUMO pathway is required for cancer cell proliferation in vitro and tumor growth in vivo, implicating the SUMO pathway as a potential cancer therapeutic target.

  5. An Arabidopsis SUMO E3 Ligase, SIZ1, Negatively Regulates Photomorphogenesis by Promoting COP1 Activity

    KAUST Repository

    Lin, Xiao-Li

    2016-04-29

    COP1 (CONSTITUTIVE PHOTOMORPHOGENIC 1), a ubiquitin E3 ligase, is a central negative regulator of photomorphogenesis. However, how COP1 activity is regulated by post-translational modifications remains largely unknown. Here we show that SUMO (small ubiquitin-like modifier) modification enhances COP1 activity. Loss-of-function siz1 mutant seedlings exhibit a weak constitutive photomorphogenic phenotype. SIZ1 physically interacts with COP1 and mediates the sumoylation of COP1. A K193R substitution in COP1 blocks its SUMO modification and reduces COP1 activity in vitro and in planta. Consistently, COP1 activity is reduced in siz1 and the level of HY5, a COP1 target protein, is increased in siz1. Sumoylated COP1 may exhibits higher transubiquitination activity than does non-sumoylated COP1, but SIZ1-mediated SUMO modification does not affect COP1 dimerization, COP1-HY5 interaction, and nuclear accumulation of COP1. Interestingly, prolonged light exposure reduces the sumoylation level of COP1, and COP1 mediates the ubiquitination and degradation of SIZ1. These regulatory mechanisms may maintain the homeostasis of COP1 activity, ensuing proper photomorphogenic development in changing light environment. Our genetic and biochemical studies identify a function for SIZ1 in photomorphogenesis and reveal a novel SUMO-regulated ubiquitin ligase, COP1, in plants.

  6. SUMO-1 Enhancing the p53-induced HepG2 Cell Apoptosis

    Institute of Scientific and Technical Information of China (English)

    LU Xingrong; YI Jilin

    2005-01-01

    Summary: In order to investigate the effect of small ubiquitin-like modifier-1 (SUMO-1) on the p53-induced HepG2 cell apoptosis, HepG2 cells were transfected by recombinant plasmids as pwtp53, pMDM2 and pSUMO-1 respectively. Western blot was employed to detect the protein expression of the transfected recombinant plasmids and the rate of apoptosis was measured by flow cytometry. The results showed that in cells transfected with pwtp53 and pwtp53+pSUMO-1, the apoptosis rate was (16.79±1.62) % and (18.15±1.36) % respectively, while transfected with pwtp53+pMDM2, the rate was decreased to (5.17±1.23) %. The apoptosis rate was (14.06±1.84) % in the cells transfected with pwtp53+pMDM2+pSUMO-1, significantly higher than that in the cells Transfected with pwtp53+pMDM2 (P<0.01). The apoptosis rates in the cells were all less than 2 % and had no significant difference among the groups. It was suggested that in the HepG2 cells, SUMO-1 can increase the apoptosis induced by wild-type p53 through binding to p53 protein, post-translational modification and inhibiting the p53 degradation by MDM2.

  7. Lamin A reassembly at the end of mitosis is regulated by its SUMO-interacting motif.

    Science.gov (United States)

    Moriuchi, Takanobu; Kuroda, Masaki; Kusumoto, Fumiya; Osumi, Takashi; Hirose, Fumiko

    2016-03-01

    Modification of proteins with small ubiquitin-related modifier (SUMO; SUMOylation) is involved in the regulation of various biological processes. Recent studies have demonstrated that noncovalent associations between SUMOylated proteins and co-operative proteins containing SUMO-interacting motifs (SIMs) are important for the spatiotemporal organization of many protein complexes. In this study, we demonstrate that interactions between lamin A, a major component of the nuclear lamina, and SUMO isoforms are dependent on one of the four SIMs (SIM3) resided in lamin A polypeptide in vitro. Live cell imaging and immunofluorescence staining showed that SIM3 is required for accumulation of lamin A on the chromosomes during telophase, and subsequent evaluation of a panel of deletion mutants determined that a 156-amino acid region spanning the carboxyl-terminal Ig-fold domain of lamin A is sufficient for this accumulation. Notably, mutation of SIM3 abrogated the dephosphorylation of mitosis-specific phosphorylation at Ser-22 of lamin A, which normally occurs during telophase, and the subsequent nuclear lamina reorganization. Furthermore, expression of a conjugation-defective SUMO2 mutant, which was previously shown to inhibit endogenous SUMOylation in a dominant-negative manner, also impaired the accumulation of wild type lamin A on telophase chromosomes. These findings suggest that interactions between SIM3 of lamin A and a putative SUMO2-modified protein plays an important role in the reorganization of the nuclear lamina at the end of mitosis.

  8. Production of FMDV virus-like particles by a SUMO fusion protein approach in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Liang Shu-Mei

    2009-08-01

    Full Text Available Abstract Virus-like particles (VLPs are formed by the self-assembly of envelope and/or capsid proteins from many viruses. Some VLPs have been proven successful as vaccines, and others have recently found applications as carriers for foreign antigens or as scaffolds in nanoparticle biotechnology. However, production of VLP was usually impeded due to low water-solubility of recombinant virus capsid proteins. Previous studies revealed that virus capsid and envelope proteins were often posttranslationally modified by SUMO in vivo, leading into a hypothesis that SUMO modification might be a common mechanism for virus proteins to retain water-solubility or prevent improper self-aggregation before virus assembly. We then propose a simple approach to produce VLPs of viruses, e.g., foot-and-mouth disease virus (FMDV. An improved SUMO fusion protein system we developed recently was applied to the simultaneous expression of three capsid proteins of FMDV in E. coli. The three SUMO fusion proteins formed a stable heterotrimeric complex. Proteolytic removal of SUMO moieties from the ternary complexes resulted in VLPs with size and shape resembling the authentic FMDV. The method described here can also apply to produce capsid/envelope protein complexes or VLPs of other disease-causing viruses.

  9. Lamin A/C mutants disturb sumo1 localization and sumoylation in vitro and in vivo.

    Directory of Open Access Journals (Sweden)

    Émilie Boudreau

    Full Text Available A-type lamins A and C are nuclear intermediate filament proteins in which mutations have been implicated in multiple disease phenotypes commonly known as laminopathies. A few studies have implicated sumoylation in the regulation of A-type lamins. Sumoylation is a post-translational protein modification that regulates a wide range of cellular processes through the attachment of small ubiquitin-related modifier (sumo to various substrates. Here we showed that laminopathy mutants result in the mislocalization of sumo1 both in vitro (C2C12 cells overexpressing mutant lamins A and C and in vivo (primary myoblasts and myopathic muscle tissue from the Lmna(H222P/H222P mouse model. In C2C12 cells, we showed that the trapping of sumo1 in p.Asp192Gly, p.Gln353Lys, and p.Arg386Lys aggregates of lamin A/C correlated with an increased steady-state level of sumoylation. However, lamin A and C did not appear to be modified by sumo1. Our results suggest that mutant lamin A/C alters the dynamics of sumo1 and thus misregulation of sumoylation may be contributing to disease progression in laminopathies.

  10. Functional characterization of a SUMO deconjugating protease of Plasmodium falciparum using newly identified small molecule inhibitors.

    Science.gov (United States)

    Ponder, Elizabeth L; Albrow, Victoria E; Leader, Brittany A; Békés, Miklós; Mikolajczyk, Jowita; Fonović, Urša Pečar; Shen, Aimee; Drag, Marcin; Xiao, Junpeng; Deu, Edgar; Campbell, Amy J; Powers, James C; Salvesen, Guy S; Bogyo, Matthew

    2011-06-24

    Small ubiquitin-related modifier (SUMO) is implicated in the regulation of numerous biological processes including transcription, protein localization, and cell cycle control. Protein modification by SUMO is found in Plasmodium falciparum; however, its role in the regulation of the parasite life cycle is poorly understood. Here we describe functional studies of a SUMO-specific protease (SENP) of P. falciparum, PfSENP1 (PFL1635w). Expression of the catalytic domain of PfSENP1 and biochemical profiling using a positional scanning substrate library demonstrated that this protease has unique cleavage sequence preference relative to the human SENPs. In addition, we describe a class of small molecule inhibitors of this protease. The most potent lead compound inhibited both recombinant PfSENP1 activity and P. falciparum replication in infected human blood. These studies provide valuable new tools for the study of SUMOylation in P. falciparum. Copyright © 2011 Elsevier Ltd. All rights reserved.

  11. Proteins with two SUMO-like domains in chromatin-associated complexes: The RENi (Rad60-Esc2-NIP45 family

    Directory of Open Access Journals (Sweden)

    Eisenhaber Birgit

    2005-02-01

    Full Text Available Abstract Background Post-translational modification by Small Ubiquitin-like Modifiers (SUMO has been implicated in protein targeting, in the maintenance of genomic integrity and in transcriptional control. But the specific molecular effects of SUMO modification on many target proteins remain to be elucidated. Recent findings point at the importance of SUMO-mediated histone NAD-dependent deacetylase (HDAC recruitment in transcriptional regulation. Results We describe the RENi family of SUMO-like domain proteins (SDP with the unique feature of typically containing two carboxy-terminal SUMO-like domains. Using sequence analytic evidence, we collect family members from animals, fungi and plants, most prominent being yeast Rad60, Esc2 and mouse NIP45 http://mendel.imp.univie.ac.at/SEQUENCES/reni/. Different proteins of the novel family are known to interact directly with histone NAD-dependent deacetylases (HDACs, structural maintenance of chromosomes (SMC proteins, and transcription factors. In particular, the highly non-trivial designation of the first of the two successive SUMO-domains in non-plant RENi provides a rationale for previously published functionally impaired mutant variants. Conclusions Till now, SUMO-like proteins have been studied exclusively in the context of their covalent conjugation to target proteins. Here, we present the exciting possibility that SUMO domain proteins, similarly to ubiquitin modifiers, have also evolved in a second line – namely as multi-domain proteins that are non-covalently attached to their target proteins. We suggest that the SUMO stable fusion proteins of the RENi family, which we introduce in this work, might mimic SUMO and share its interaction motifs (in analogy to the way that ubiquitin-like domains mimic ubiquitin. This presumption is supported by parallels in the spectrum of modified or bound proteins e.g. transcription factors and chromatin-associated proteins and in the recruitment of HDAC-activity.

  12. Intense Resistance Exercise Promotes the Acute and Transient Nuclear Translocation of Small Ubiquitin-Related Modifier (SUMO-1 in Human Myofibres

    Directory of Open Access Journals (Sweden)

    Sebastian Gehlert

    2016-04-01

    Full Text Available Protein sumoylation is a posttranslational modification triggered by cellular stress. Because general information concerning the role of small ubiquitin-related modifier (SUMO proteins in adult skeletal muscle is sparse, we investigated whether SUMO-1 proteins will be subjected to time-dependent changes in their subcellular localization in sarcoplasmic and nuclear compartments of human type I and II skeletal muscle fibers in response to acute stimulation by resistance exercise (RE. Skeletal muscle biopsies were taken at baseline (PRE, 15, 30, 60, 240 min and 24 h post RE from 6 male subjects subjected to a single bout of one-legged knee extensions. SUMO-1 localization was determined via immunohistochemistry and confocal laser microscopy. At baseline SUMO-1 was localized in perinuclear regions of myonuclei. Within 15 and up to 60 min post exercise, nuclear SUMO-1 localization was significantly increased (p < 0.01, declining towards baseline levels within 240 min post exercise. Sarcoplasmic SUMO-1 localization was increased at 15 min post exercise in type I and up to 30 min post RE in type II myofibres. The changing localization of SUMO-1 proteins acutely after intense muscle contractions points to a role for SUMO proteins in the acute regulation of the skeletal muscle proteome after exercise.

  13. Analysis of the SUMO2 Proteome during HSV-1 Infection.

    Directory of Open Access Journals (Sweden)

    Elizabeth Sloan

    2015-07-01

    Full Text Available Covalent linkage to members of the small ubiquitin-like (SUMO family of proteins is an important mechanism by which the functions of many cellular proteins are regulated. Sumoylation has roles in the control of protein stability, activity and localization, and is involved in the regulation of transcription, gene expression, chromatin structure, nuclear transport and RNA metabolism. Sumoylation is also linked, both positively and negatively, with the replication of many different viruses both in terms of modification of viral proteins and modulation of sumoylated cellular proteins that influence the efficiency of infection. One prominent example of the latter is the widespread reduction in the levels of cellular sumoylated species induced by herpes simplex virus type 1 (HSV-1 ubiquitin ligase ICP0. This activity correlates with relief from intrinsic immunity antiviral defence mechanisms. Previous work has shown that ICP0 is selective in substrate choice, with some sumoylated proteins such the promyelocytic leukemia protein PML being extremely sensitive, while RanGAP is completely resistant. Here we present a comprehensive proteomic analysis of changes in the cellular SUMO2 proteome during HSV-1 infection. Amongst the 877 potentially sumoylated species detected, we identified 124 whose abundance was decreased by a factor of 3 or more by the virus, several of which were validated by western blot and expression analysis. We found many previously undescribed substrates of ICP0 whose degradation occurs by a range of mechanisms, influenced or not by sumoylation and/or the SUMO2 interaction motif within ICP0. Many of these proteins are known or are predicted to be involved in the regulation of transcription, chromatin assembly or modification. These results present novel insights into mechanisms and host cell proteins that might influence the efficiency of HSV-1 infection.

  14. PIAS1 is a GATA4 SUMO ligase that regulates GATA4-dependent intestinal promoters independent of SUMO ligase activity and GATA4 sumoylation.

    Directory of Open Access Journals (Sweden)

    Narasimhaswamy S Belaguli

    Full Text Available GATA4 confers cell type-specific gene expression on genes expressed in cardiovascular, gastro-intestinal, endocrine and neuronal tissues by interacting with various ubiquitous and cell-type-restricted transcriptional regulators. By using yeast two-hybrid screening approach, we have identified PIAS1 as an intestine-expressed GATA4 interacting protein. The physical interaction between GATA4 and PIAS1 was confirmed in mammalian cells by coimmunoprecipitation and two-hybrid analysis. The interacting domains were mapped to the second zinc finger and the adjacent C-terminal basic region of GATA4 and the RING finger and the adjoining C-terminal 60 amino acids of PIAS1. PIAS1 and GATA4 synergistically activated IFABP and SI promoters but not LPH promoters suggesting that PIAS1 differentially activates GATA4 targeted promoters. In primary murine enterocytes PIAS1 was recruited to the GATA4-regulated IFABP promoter. PIAS1 promoted SUMO-1 modification of GATA4 on lysine 366. However, sumoylation was not required for the nuclear localization and stability of GATA4. Further, neither GATA4 sumoylation nor the SUMO ligase activity of PIAS1 was required for coactivation of IFABP promoter by GATA4 and PIAS1. Together, our results demonstrate that PIAS1 is a SUMO ligase for GATA4 that differentially regulates GATA4 transcriptional activity independent of SUMO ligase activity and GATA4 sumoylation.

  15. Functional Crosstalk between the PP2A and SUMO Pathways Revealed by Analysis of STUbL Suppressor, razor 1-1.

    Directory of Open Access Journals (Sweden)

    Minghua Nie

    2016-07-01

    Full Text Available Posttranslational modifications (PTMs provide dynamic regulation of the cellular proteome, which is critical for both normal cell growth and for orchestrating rapid responses to environmental stresses, e.g. genotoxins. Key PTMs include ubiquitin, the Small Ubiquitin-like MOdifier SUMO, and phosphorylation. Recently, SUMO-targeted ubiquitin ligases (STUbLs were found to integrate signaling through the SUMO and ubiquitin pathways. In general, STUbLs are recruited to target proteins decorated with poly-SUMO chains to ubiquitinate them and drive either their extraction from protein complexes, and/or their degradation at the proteasome. In fission yeast, reducing or preventing the formation of SUMO chains can circumvent the essential and DNA damage response functions of STUbL. This result indicates that whilst some STUbL "targets" have been identified, the crucial function of STUbL is to antagonize SUMO chain formation. Herein, by screening for additional STUbL suppressors, we reveal crosstalk between the serine/threonine phosphatase PP2A-Pab1B55 and the SUMO pathway. A hypomorphic Pab1B55 mutant not only suppresses STUbL dysfunction, but also mitigates the phenotypes associated with deletion of the SUMO protease Ulp2, or mutation of the STUbL cofactor Rad60. Together, our results reveal a novel role for PP2A-Pab1B55 in modulating SUMO pathway output, acting in parallel to known critical regulators of SUMOylation homeostasis. Given the broad evolutionary functional conservation of the PP2A and SUMO pathways, our results could be relevant to the ongoing attempts to therapeutically target these factors.

  16. Sumo-dependent substrate targeting of the SUMO protease Ulp1

    Directory of Open Access Journals (Sweden)

    Westerbeck Jason W

    2011-10-01

    Full Text Available Abstract Background In the yeast Saccharomyces cerevisiae, the essential small ubiquitin-like modifier (SUMO protease Ulp1 is responsible for both removing SUMO/Smt3 from specific target proteins and for processing precursor SUMO into its conjugation-competent form. Ulp1 localizes predominantly to nuclear pore complexes but has also been shown to deconjugate sumoylated septins at the bud-neck of dividing cells. How Ulp1 is directed to bud-neck localized septins and other cytoplasmic deconjugation targets is not well understood. Results Using a structure/function approach, we set out to elucidate features of Ulp1 that are required for substrate targeting. To aid our studies, we took advantage of a catalytically inactive mutant of Ulp1 that is greatly enriched at the septin ring of dividing yeast cells. We found that the localization of Ulp1 to the septins requires both SUMO and specific structural features of Ulp1's catalytic domain. Our analysis identified a 218-amino acid, substrate-trapping mutant of the catalytic domain of Ulp1, Ulp1(3(C580S, that is necessary and sufficient for septin localization. We also used the targeting and SUMO-binding properties of Ulp1(3(C580S to purify Smt3-modified proteins from cell extracts. Conclusions Our study provides novel insights into how the Ulp1 SUMO protease is actively targeted to its substrates in vivo and in vitro. Furthermore, we found that a substrate-trapping Ulp1(3(C580S interacts robustly with human SUMO1, SUMO2 and SUMO2 chains, making it a potentially useful tool for the analysis and purification of SUMO-modified proteins.

  17. Elevated salicylic acid levels conferred by increased expression of ISOCHORISMATE SYNTHASE 1 contribute to hyperaccumulation of SUMO1 conjugates in the Arabidopsis mutant early in short days 4.

    Science.gov (United States)

    Villajuana-Bonequi, Mitzi; Elrouby, Nabil; Nordström, Karl; Griebel, Thomas; Bachmair, Andreas; Coupland, George

    2014-07-01

    Post-translational modification of proteins by attachment of small ubiquitin-like modifier (SUMO) is essential for plant growth and development. Mutations in the SUMO protease early in short days 4 (ESD4) cause hyperaccumulation of conjugates formed between SUMO and its substrates, and phenotypically are associated with extreme early flowering and impaired growth. We performed a suppressor mutagenesis screen of esd4 and identified a series of mutants called suppressor of esd4 (sed), which delay flowering, enhance growth and reduce hyperaccumulation of SUMO conjugates. Genetic mapping and genome sequencing indicated that one of these mutations (sed111) is in the gene salicylic acid induction-deficient 2 (SID2), which encodes ISOCHORISMATE SYNTHASE I, an enzyme required for biosynthesis of salicylic acid (SA). Analyses showed that compared with wild-type plants, esd4 contains higher levels of SID2 mRNA and about threefold more SA, whereas sed111 contains lower SA levels. Other sed mutants also contain lower SA levels but are not mutant for SID2, although most reduce SID2 mRNA levels. Therefore, higher SA levels contribute to the small size, early flowering and elevated SUMO conjugate levels of esd4. Our results support previous data indicating that SUMO homeostasis influences SA biosynthesis in wild-type plants, and also demonstrate that elevated levels of SA strongly increase the abundance of SUMO conjugates.

  18. A comprehensive compilation of SUMO proteomics

    DEFF Research Database (Denmark)

    Hendriks, Ivo A; Vertegaal, Alfred C O

    2016-01-01

    Small ubiquitin-like modifiers (SUMOs) are essential for the regulation of several cellular processes and are potential therapeutic targets owing to their involvement in diseases such as cancer and Alzheimer disease. In the past decade, we have witnessed a rapid expansion of proteomic approaches...... for identifying sumoylated proteins, with recent advances in detecting site-specific sumoylation. In this Analysis, we combined all human SUMO proteomics data currently available into one cohesive database. We provide proteomic evidence for sumoylation of 3,617 proteins at 7,327 sumoylation sites, and insight...

  19. SUMO-conjugating enzyme E2 UBC9 mediates viral immediate-early protein SUMOylation in crayfish to facilitate reproduction of white spot syndrome virus.

    Science.gov (United States)

    Chen, An-Jing; Gao, Lu; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-01-01

    Successful viruses have evolved superior strategies to escape host defenses or exploit host biological pathways. Most of the viral immediate-early (ie) genes are essential for viral infection and depend solely on host proteins; however, the molecular mechanisms are poorly understood. In this study, we focused on the modification of viral IE proteins by the crayfish small ubiquitin-related modifier (SUMO) and investigated the role of SUMOylation during the viral life cycle. SUMO and SUMO ubiquitin-conjugating enzyme 9 (UBC9) involved in SUMOylation were identified in red swamp crayfish (Procambarus clarkii). Both SUMO and UBC9 were upregulated in crayfish challenged with white spot syndrome virus (WSSV). Replication of WSSV genes increased in crayfish injected with recombinant SUMO or UBC9, but injection of mutant SUMO or UBC9 protein had no effect. Subsequently, we analyzed the mechanism by which crayfish SUMOylation facilitates WSSV replication. Crayfish UBC9 bound to all three WSSV IE proteins tested, and one of these IE proteins (WSV051) was covalently modified by SUMO in vitro. The expression of viral ie genes was affected and that of late genes was significantly inhibited in UBC9-silenced or SUMO-silenced crayfish, and the inhibition effect was rescued by injection of recombinant SUMO or UBC9. The results of this study demonstrate that viral IE proteins can be modified by crayfish SUMOylation, prompt the expression of viral genes, and ultimately benefit WSSV replication. Understanding of the mechanisms by which viruses exploit host components will greatly improve our knowledge of the virus-host "arms race" and contribute to the development of novel methods against virulent viruses.

  20. USP7/HAUSP: A SUMO deubiquitinase at the heart of DNA replication.

    Science.gov (United States)

    Smits, Veronique A J; Freire, Raimundo

    2016-09-01

    DNA replication is both highly conserved and controlled. Problematic DNA replication can lead to genomic instability and therefore carcinogenesis. Numerous mechanisms work together to achieve this tight control and increasing evidence suggests that post-translational modifications (phosphorylation, ubiquitination, SUMOylation) of DNA replication proteins play a pivotal role in this process. Here we discuss such modifications in the light of a recent article that describes a novel role for the deubiquitinase (DUB) USP7/HAUSP in the control of DNA replication. USP7 achieves this function by an unusual and novel mechanism, namely deubiquitination of SUMOylated proteins at the replication fork, making USP7 also a SUMO DUB (SDUB). This work extends previous observations of increased levels of SUMO and low levels of ubiquitin at the on-going replication fork. Here, we discuss this novel study, its contribution to the DNA replication and genomic stability field and what questions arise from this work.

  1. Cooperativity of the SUMO and Ubiquitin Pathways in Genome Stability

    Directory of Open Access Journals (Sweden)

    Minghua Nie

    2016-02-01

    Full Text Available Covalent attachment of ubiquitin (Ub or SUMO to DNA repair proteins plays critical roles in maintaining genome stability. These structurally related polypeptides can be viewed as distinct road signs, with each being read by specific protein interaction motifs. Therefore, via their interactions with selective readers in the proteome, ubiquitin and SUMO can elicit distinct cellular responses, such as directing DNA lesions into different repair pathways. On the other hand, through the action of the SUMO-targeted ubiquitin ligase (STUbL family proteins, ubiquitin and SUMO can cooperate in the form of a hybrid signal. These mixed SUMO-ubiquitin chains recruit “effector” proteins such as the AAA+ ATPase Cdc48/p97-Ufd1-Npl4 complex that contain both ubiquitin and SUMO interaction motifs. This review will summarize recent key findings on collaborative and distinct roles that ubiquitin and SUMO play in orchestrating DNA damage responses.

  2. Uncovering SUMOylation Dynamics during Cell-Cycle Progression Reveals FoxM1 as a Key Mitotic SUMO Target Protein

    DEFF Research Database (Denmark)

    Schimmel, Joost; Eifler, Karolin; Sigurdsson, Jón Otti;

    2014-01-01

    Loss of small ubiquitin-like modification (SUMOylation) in mice causes genomic instability due to the missegregation of chromosomes. Currently, little is known about the identity of relevant SUMO target proteins that are involved in this process and about global SUMOylation dynamics during cell-c...

  3. Wrestling with Chromosomes: The Roles of SUMO During Meiosis.

    Science.gov (United States)

    Nottke, Amanda C; Kim, Hyun-Min; Colaiácovo, Monica P

    2017-01-01

    Meiosis is a specialized form of cell division required for the formation of haploid gametes and therefore is essential for successful sexual reproduction. Various steps are exquisitely coordinated to ensure accurate chromosome segregation during meiosis, thereby promoting the formation of haploid gametes from diploid cells. Recent studies are demonstrating that an important form of regulation during meiosis is exerted by the post-translational protein modification known as sumoylation. Here, we review and discuss the various critical steps of meiosis in which SUMO-mediated regulation has been implicated thus far. These include the maintenance of meiotic centromeric heterochromatin , meiotic DNA double-strand break repair and homologous recombination, centromeric coupling, and the assembly of a proteinaceous scaffold between homologous chromosomes known as the synaptonemal complex.

  4. Proteome-wide screens for small ubiquitin-like modifier (SUMO) substrates identify Arabidopsis proteins implicated in diverse biological processes

    National Research Council Canada - National Science Library

    Nabil Elrouby; George Coupland

    2010-01-01

    .... Despite this, little is known about SUMO targets in plants. Here we identified 238 Arabidopsis proteins as potential SUMO substrates because they interacted with SUMO-conjugating enzyme and/or SUMO protease (ESD4...

  5. Non-native Conformers of Cystic Fibrosis Transmembrane Conductance Regulator NBD1 Are Recognized by Hsp27 and Conjugated to SUMO-2 for Degradation.

    Science.gov (United States)

    Gong, Xiaoyan; Ahner, Annette; Roldan, Ariel; Lukacs, Gergely L; Thibodeau, Patrick H; Frizzell, Raymond A

    2016-01-22

    A newly identified pathway for selective degradation of the common mutant of the cystic fibrosis transmembrane conductance regulator (CFTR), F508del, is initiated by binding of the small heat shock protein, Hsp27. Hsp27 collaborates with Ubc9, the E2 enzyme for protein SUMOylation, to selectively degrade F508del CFTR via the SUMO-targeted ubiquitin E3 ligase, RNF4 (RING finger protein 4) (1). Here, we ask what properties of CFTR are sensed by the Hsp27-Ubc9 pathway by examining the ability of NBD1 (locus of the F508del mutation) to mimic the disposal of full-length (FL) CFTR. Similar to FL CFTR, F508del NBD1 expression was reduced 50-60% by Hsp27; it interacted preferentially with the mutant and was modified primarily by SUMO-2. Mutation of the consensus SUMOylation site, Lys(447), obviated Hsp27-mediated F508del NBD1 SUMOylation and degradation. As for FL CFTR and NBD1 in vivo, SUMO modification using purified components in vitro was greater for F508del NBD1 versus WT and for the SUMO-2 paralog. Several findings indicated that Hsp27-Ubc9 targets the SUMOylation of a transitional, non-native conformation of F508del NBD1: (a) its modification decreased as [ATP] increased, reflecting stabilization of the nucleotide-binding domain by ligand binding; (b) a temperature-induced increase in intrinsic fluorescence, which reflects formation of a transitional NBD1 conformation, was followed by its SUMO modification; and (c) introduction of solubilizing or revertant mutations to stabilize F508del NBD1 reduced its SUMO modification. These findings indicate that the Hsp27-Ubc9 pathway recognizes a non-native conformation of mutant NBD1, which leads to its SUMO-2 conjugation and degradation by the ubiquitin-proteasome system.

  6. Mass Spectral Enhanced Detection of Ubls Using SWATH Acquisition: MEDUSA—Simultaneous Quantification of SUMO and Ubiquitin-Derived Isopeptides

    Science.gov (United States)

    Griffiths, John R.; Chicooree, Navin; Connolly, Yvonne; Neffling, Milla; Lane, Catherine S.; Knapman, Thomas; Smith, Duncan L.

    2014-05-01

    Protein modification by ubiquitination and SUMOylation occur throughout the cell and are responsible for numerous cellular functions such as apoptosis, DNA replication and repair, and gene transcription. Current methods for the identification of such modifications using mass spectrometry predominantly rely upon tryptic isopeptide tag generation followed by database searching with in vitro genetic mutation of SUMO routinely required. We have recently described a novel approach to ubiquitin and SUMO modification detection based upon the diagnostic a' and b' ions released from the isopeptide tags upon collision-induced dissociation of reductively methylated Ubl isopeptides (RUbI) using formaldehyde. Here, we significantly extend those studies by combining data-independent acquisition (DIA) with alternative labeling reagents to improve diagnostic ion coverage and enable relative quantification of modified peptides from both MS and MS/MS signals. Model synthetic ubiquitin and SUMO-derived isopeptides were labeled with mTRAQ reagents (Δ0, Δ4, and Δ8) and subjected to LC-MS/MS with SWATH acquisition. Novel diagnostic ions were generated upon CID, which facilitated the selective detection of these modified peptides. Simultaneous MS-based and MS/MS-based relative quantification was demonstrated for both Ub and SUMO-derived isopeptides across three channels in a background of mTRAQ-labeled Escherichia coli digest.

  7. Analysis list: SUMO2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available SUMO2 Blood,Prostate + hg19 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/target/...SUMO2.1.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/target/SUMO2.5.tsv http://dbarchive.biosciencedb...c.jp/kyushu-u/hg19/target/SUMO2.10.tsv http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/SUMO2.Blood.tsv,http://dbarchive.bioscien...cedbc.jp/kyushu-u/hg19/colo/SUMO2.Prostate.tsv http://dbarchive.bioscience...dbc.jp/kyushu-u/hg19/colo/Blood.gml,http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/colo/Prostate.gml ...

  8. SUMO modulation of protein aggregation and degradation

    Directory of Open Access Journals (Sweden)

    Marco Feligioni

    2015-09-01

    Full Text Available Small ubiquitin-like modifier (SUMO conjugation and binding to target proteins regulate a wide variety of cellular pathways. The functional aspects of SUMOylation include changes in protein-protein interactions, intracellular trafficking as well as protein aggregation and degradation. SUMO has also been linked to specialized cellular pathways such as neuronal development and synaptic transmission. In addition, SUMOylation is associated with neurological diseases associated with abnormal protein accumulations. SUMOylation of the amyloid and tau proteins involved in Alzheimer's disease and other tauopathies may contribute to changes in protein solubility and proteolytic processing. Similar events have been reported for α-synuclein aggregates found in Parkinson's disease, polyglutamine disorders such as Huntington's disease as well as protein aggregates found in amyotrophic lateral sclerosis (ALS. This review provides a detailed overview of the impact SUMOylation has on the etiology and pathology of these related neurological diseases.

  9. SUMO: Solar Ultraviolet Monitor and Ozone Nanosatellite

    Science.gov (United States)

    Damé, L.; Meftah, M.; Irbah, A.; Hauchecorne, A.; Keckhut, P.; Sarkissian, A.; Godin-Beekman, S.; Rogers, D. J.; Bove, P.; Lagage, P. O.; DeWitte, S.

    2014-12-01

    SUMO is an innovative proof-of-concept nanosatellite aiming to measure on the same platform the different components of the Earth radiation budget (ERB), the solar energy input and the energy reemitted at the top of the Earth atmosphere, with a particular focus on the far UV (FUV) part of the spectrum and on the ozone layer. The FUV is the only wavelength band with energy absorbed in the high atmosphere (stratosphere), in the ozone (Herzberg continuum, 200-220 nm) and oxygen bands, and its high variability is most probably at the origin of a climate influence (UV affects stratospheric dynamics and temperatures, altering interplanetary waves and weather patterns both poleward and downward to the lower stratosphere and tropopause). A simultaneous observation of incoming FUV and ozone production would bring an invaluable information on this process of solar-climate forcing. Space instruments have already measured the different components of the ERB but this is the first time that all instruments will operate on the same platform. This characteristic by itself guarantees original scientific results. SUMO is a 3.6 kg, 3W, 10x10x30 cm3 nanosatellite ("3U"), with a "1U" payload of definition has been completed (platform and payload AIT are possible in 24 months). SUMO is proposed for the nanosatellite program of Polytechnic School and CNES (following QB50) for a flight in 2018. Follow-up is 2 fold: on one part more complete measurements using SUMO miniaturized instruments on a larger satellite; on the other part, increase of the coverage in local time and latitude using a constellation of SUMO nanosatellites around the Earth to further geolocalize the Sun influence on our planet. Nanosatellites, with cost and risk limited, are also excellent platforms to evaluate technologies for future missions, e.g. nanotechnology ZnO protection barriers to limit contamination from solar panels in the UV and reduce reflection losses in the visible, or MgZnO solar blind detectors (R

  10. Puromycin induces SUMO and ubiquitin redistribution upon proteasome inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Hotaru [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Saitoh, Hisato, E-mail: hisa@kumamoto-u.ac.jp [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto (Japan)

    2016-07-29

    We have previously reported the co-localization of O-propargyl-puromycin (OP-Puro) with SUMO-2/3 and ubiquitin at promyelocytic leukemia-nuclear bodies (PML-NBs) in the presence of the proteasome inhibitor MG132, implying a role for the ubiquitin family in sequestering OP-puromycylated immature polypeptides to the nucleus during impaired proteasome activity. Here, we found that as expected puromycin induced SUMO-1/2/3 accumulation with ubiquitin at multiple nuclear foci in HeLa cells when co-exposed to MG132. Co-administration of puromycin and MG132 also facilitated redistribution of PML and the SUMO-targeted ubiquitin ligase RNF4 concurrently with SUMO-2/3. As removal of the drugs from the medium led to disappearance of the SUMO-2/3-ubiquitin nuclear foci, our findings indicated that nuclear assembly/disassembly of SUMO-2/3 and ubiquitin was pharmacologically manipulable, supporting our previous observation on OP-Puro, which predicted the ubiquitin family function in sequestrating aberrant proteins to the nucleus. -- Highlights: •Puromycin exhibits the O-propargyl-puromycin effect. •Puromycin induces SUMO redistribution upon proteasome inhibition. •Ubiquitin and RNF4 accumulate at PML-nuclear bodies with SUMO-2/3. •The ubiquitin family may function in nuclear sequestration of immature proteins.

  11. Global SUMO proteome responses guide gene regulation, mRNA biogenesis, and plant stress responses.

    NARCIS (Netherlands)

    M.J. Mazur; H.A. van den Burg

    2012-01-01

    Small Ubiquitin-like MOdifier (SUMO) is a key regulator of abiotic stress, disease resistance, and development in plants. The identification of >350 plant SUMO targets has revealed many processes modulated by SUMO and potential consequences of SUMO on its targets. Importantly, highly related protein

  12. Sentrin/SUMO specific proteases as novel tissue-selective modulators of vitamin D receptor-mediated signaling.

    Directory of Open Access Journals (Sweden)

    Wai-Ping Lee

    Full Text Available Vitamin D receptor (VDR is a substrate for modification with small ubiquitin-like modifier (SUMO. To further assess the role of reversible SUMOylation within the vitamin D hormonal response, we evaluated the effects of sentrin/SUMO-specific proteases (SENPs that can function to remove small ubiquitin-like modifier (SUMO from target proteins upon the activities of VDR and related receptors. We report that SENP1 and SENP2 strikingly potentiate ligand-mediated transactivation of VDR and also its heterodimeric partner, retinoid X receptor (RXRα with depletion of cellular SENP1 significantly diminishing the hormonal responsiveness of the endogenous vitamin D target gene CYP24A1. We find that SENP-directed modulation of VDR activity is cell line-dependent, achieving potent modulatory effects in Caco-2 and HEK-293 cells, while in MCF-7 cells the vitamin D signal is unaffected by any tested SENP. In support of their function as novel modulators of the vitamin D hormonal pathway we demonstrate that both SENP1 and SENP2 can interact with VDR and reverse its modification with SUMO2. In a preliminary analysis we identify lysine 91, a residue known to be critical for formation and DNA binding of the VDR-RXR heterodimer, as a minor SUMO acceptor site within VDR. In combination, our results support a repressor function for SUMOylation of VDR and reveal SENPs as a novel class of VDR/RXR co-regulatory protein that significantly modulate the vitamin D response and which could also have important impact upon the functionality of both RXR-containing homo and heterodimers.

  13. The small ubiquitin-like modifier (SUMO) is required for gonadal and uterine-vulval morphogenesis in Caenorhabditis elegans.

    Science.gov (United States)

    Broday, Limor; Kolotuev, Irina; Didier, Christine; Bhoumik, Anindita; Gupta, Bhagwati P; Sternberg, Paul W; Podbilewicz, Benjamin; Ronai, Ze'ev

    2004-10-01

    The small ubiquitin-like modifier (SUMO) modification alters the subcellular distribution and function of its substrates. Here we show the major role of SUMO during the development of the Caenorhabditis elegans reproductive system. smo-1 deletion mutants develop into sterile adults with abnormal somatic gonad, germ line, and vulva. SMO-1::GFP reporter is highly expressed in the somatic reproductive system. smo-1 animals lack a vulval-uterine connection as a result of impaired ventral uterine pi-cell differentiation and anchor cell fusion. Mutations in the LIN-11 LIM domain transcription factor lead to a uterine phenotype that resembles the smo-1 phenotype. LIN-11 is sumoylated, and its sumoylation is required for its activity during uterine morphogenesis. Expression of a SUMO-modified LIN-11 in the smo-1 background partially rescued pi-cell differentiation and retained LIN-11 in nuclear bodies. Thus, our results identify the reproductive system as the major SUMO target during postembryonic development and highlight LIN-11 as a physiological substrate whose sumoylation is associated with the formation of a functional vulval-uterine connection.

  14. Real Estate in the DNA Damage Response: Ubiquitin and SUMO Ligases Home in on DNA Double-Strand Breaks.

    Science.gov (United States)

    Dantuma, Nico P; Pfeiffer, Annika

    2016-01-01

    Ubiquitin and the ubiquitin-like modifier SUMO are intimately connected with the cellular response to various types of DNA damage. A striking feature is the local accumulation of these proteinaceous post-translational modifications in the direct vicinity to DNA double-strand breaks, which plays a critical role in the formation of ionizing radiation-induced foci. The functional significance of these modifications is the coordinated recruitment and removal of proteins involved in DNA damage signaling and repair in a timely manner. The central orchestrators of these processes are the ubiquitin and SUMO ligases that are responsible for accurately tagging a broad array of chromatin and chromatin-associated proteins thereby changing their behavior or destination. Despite many differences in the mode of action of these enzymes, they share some striking features that are of direct relevance for their function in the DNA damage response. In this review, we outline the molecular mechanisms that are responsible for the recruitment of ubiquitin and SUMO ligases and discuss the importance of chromatin proximity in this process.

  15. SUMO E3 Ligase AtMMS21 Regulates Drought Tolerance in Arabidopsis thaliana

    Institute of Scientific and Technical Information of China (English)

    Shengchun Zhang; Yanli Qi; Ming Liu; ChengweiYang

    2013-01-01

    Post-translational modifications of proteins by small ubiquitin-like modifiers (SUMOs) play crucial roles in plant growth and development,and in stress responses.The MMS21 is a newly-identified Arabidopsis thaliana L.SUMO E3 ligase gene aside from the SIZ1,and its function requires further elucidation.Here,we show that MMS21 deficient plants display improved drought tolerance,and constitutive expression of MMS21 reduces drought tolerance.The expression of MMS21 was reduced by abscisic acid (ABA),polyethylene glycol (PEG) or drought stress.Under drought conditions,mms21 mutants showed the highest survival rate and the slowest water loss,and accumulated a higher level of free proline compared to wild-type (WT) and MMS21 over-expression plants.Stomatal aperture,seed germination and cotyledon greening analysis indicated that mms21 was hypersensitive to ABA.Molecular genetic analysis revealed that MMS21 deficiency led to elevated expression of a series of ABA-mediated stress-responsive genes,including COR15A,RD22,and P5CS1 The ABA and drought-induced stress-responsive genes,including RAB18,RD29A and RD29B,were inhibited by constitutive expression of MMS21.Moreover,ABA-induced accumulation of SUMO-protein conjugates was blocked in the mms21 mutant.We thus conclude that MMS21 plays a role in the drought stress response,likely through regulation of gene expression in an ABA-dependent pathway.

  16. Global SUMO proteome responses guide gene regulation, mRNA biogenesis, and plant stress responses

    Directory of Open Access Journals (Sweden)

    Magdalena eMazur

    2012-09-01

    Full Text Available Small-ubiquitin-like MOdifier (SUMO is a key regulator of abiotic stress, disease resistance and development in plants. The identification of >350 plant SUMO targets has revealed many processes modulated by SUMO and potential consequences of SUMO on its targets. Importantly, highly related proteins are SUMO-modified in plants, yeast, and metazoans. Overlapping SUMO targets include heat-shock proteins, transcription regulators, histones, histone-modifying enzymes, proteins involved in DNA damage repair, but also proteins involved in mRNA biogenesis and nucleo-cytoplasmic transport. Proteomics studies indicate key roles for SUMO in gene repression by controlling histone (deacetylation activity at genomic loci. The responsible heavily sumoylated transcriptional repressor complexes are recruited by EAR (Ethylene-responsive element binding factor [ERF]-associated Amphiphilic Repression-motif containing transcription factors in plants. These transcription factors are not necessarily themselves a SUMO target. Conversely, SUMO acetylation prevents binding of downstream partners by preventing binding of SIMs (SUMO-interaction peptide motifs presents in these partners, while SUMO acetylation has emerged as mechanism to recruit specifically bromodomains; bromodomain are generally linked with gene activation. These findings strengthen the idea of a bidirectional sumo-/acetylation switch in gene regulation. Quantitative proteomics has highlighted that global sumoylation provides a dynamic response to protein damage involving SUMO chain-mediated protein degradation, but also SUMO E3 ligase-dependent transcription of HSP (Heat-shock protein genes. With these insights in SUMO function and novel technical advancements, we can now study SUMO dynamics in responses to (abiotic stress in plants.

  17. Elg1, the major subunit of an alternative RFC complex, interacts with SUMO-processing proteins.

    Science.gov (United States)

    Parnas, Oren; Amishay, Rona; Liefshitz, Batia; Zipin-Roitman, Adi; Kupiec, Martin

    2011-09-01

    PCNA is a homotrimeric ring with important roles in DNA replication and repair. PCNA is loaded and unloaded by the RFC complex, which is composed of five subunits (Rfc1-5). Three additional complexes that share with RFC the small subunits (Rfc2-5) and contain alternative large subunits were found in yeast and other eukaryotes. We have recently reported that one of these, the Elg1-RFC complex, interacts with SUMOylated PCNA and may play a role in its unloading during DNA repair. Here we report that a yeast-two-hybrid screen with the N terminus of Elg1(which interacts with SUMOylated PCNA) uncovered interactions with proteins that belong to the SUMO pathway, including Slx5 and Slx8, which form an E3 ubiquitin ligase that ubiquitinates SUMOylated proteins. Mutations in SLX5 result in a genomic instability phenotype similar to that of elg1 mutants. The physical interaction between the N terminus of Elg1 and Slx5 is mediated by poly-SUMO chains but not by PCNA modifications, and requires Siz2, but not Siz1, activity. Thus our results highlight the many important roles played by Elg1, some of which are PCNA-dependent and some PCNA-independent.

  18. Negative regulation of TLR inflammatory signaling by the SUMO-deconjugating enzyme SENP6.

    Directory of Open Access Journals (Sweden)

    Xing Liu

    Full Text Available The signaling of Toll-like receptors (TLRs induces host defense against microbial invasion. Protein posttranslational modifications dynamically shape the strength and duration of the signaling pathways. It is intriguing to explore whether de-SUMOylation could modulate the TLR signaling. Here we identified SUMO-specific protease 6 (SENP6 as an intrinsic attenuator of the TLR-triggered inflammation. Depletion of SENP6 significantly potentiated the NF-κB-mediated induction of the proinflammatory genes. Consistently, SENP6-knockdown mice were more susceptible to endotoxin-induced sepsis. Mechanistically, the small ubiquitin-like modifier 2/3 (SUMO-2/3 is conjugated onto the Lysine residue 277 of NF-κB essential modifier (NEMO/IKKγ, and this impairs the deubiquitinase CYLD to bind NEMO, thus strengthening the inhibitor of κB kinase (IKK activation. SENP6 reverses this process by catalyzing the de-SUMOylation of NEMO. Our study highlights the essential function of the SENP family in dampening TLR signaling and inflammation.

  19. Structural analysis and evolution of specificity of the SUMO UFD E1-E2 interactions

    Science.gov (United States)

    Liu, Bing; Lois, L. Maria; Reverter, David

    2017-01-01

    SUMO belongs to the ubiquitin-like family (UbL) of protein modifiers. SUMO is conserved among eukaryotes and is essential for the regulation of processes such as DNA damage repair, transcription, DNA replication and mitosis. UbL modification of proteins occurs via a specific enzymatic cascade formed by the crosstalk between the E1-activating enzyme, the E2-conjugating enzyme and the E3-ligase. An essential discrimination step in all UbL modifiers corresponds to the interaction between E1 and E2 enzymes, which is mediated by the recruitment of the E2 to the UFD domain (Ubiquitin-Fold Domain) of the E1 enzyme. To gain insights in the properties of this interface, we have compared the structures of the complexes between E1 UFD domain and E2 in human and yeast, revealing two alternative UFD platforms that interact with a conserved E2. Comparative sequence analysis of the E1 UFD domain indicates that the E2 binding region has been conserved across phylogenetic closely related species, in which higher sequence conservation can be found in the E2 binding region than in the entire UFD domain. These distinctive strategies for E1-E2 interactions through the UFD domain might be the consequence of a high selective pressure to ensure specificity of each modifier conjugation system. PMID:28165030

  20. Interferon-induced antiviral Mx1 GTPase is associated with components of the SUMO-1 system and promyelocytic leukemia protein nuclear bodies.

    Science.gov (United States)

    Engelhardt, O G; Ullrich, E; Kochs, G; Haller, O

    2001-12-10

    Mx proteins are interferon-induced large GTPases, some of which have antiviral activity against a variety of viruses. The murine Mx1 protein accumulates in the nucleus of interferon-treated cells and is active against members of the Orthomyxoviridae family, such as the influenza viruses and Thogoto virus. The mechanism by which Mx1 exerts its antiviral action is still unclear, but an involvement of undefined nuclear factors has been postulated. Using the yeast two-hybrid system, we identified cellular proteins that interact with Mx1 protein. The Mx1 interactors were mainly nuclear proteins. They included Sp100, Daxx, and Bloom's syndrome protein (BLM), all of which are known to localize to specific subnuclear domains called promyelocytic leukemia protein nuclear bodies (PML NBs). In addition, components of the SUMO-1 protein modification system were identified as Mx1-interacting proteins, namely the small ubiquitin-like modifier SUMO-1 and SAE2, which represents subunit 2 of the SUMO-1 activating enzyme. Analysis of the subcellular localization of Mx1 and some of these interacting proteins by confocal microscopy revealed a close spatial association of Mx1 with PML NBs. This suggests a role of PML NBs and SUMO-1 in the antiviral action of Mx1 and may allow us to discover novel functions of this large GTPase.

  1. Development of a space universal modular architecture (SUMO)

    Science.gov (United States)

    Collins, Bernie F.

    This concept paper proposes that the space community should develop and implement a universal standard for spacecraft modularity - to improve interoperability of spacecraft components. Pursuing a global industry consensus standard for open and modular spacecraft architecture will encourage trade, remove standards-related market barriers, and in the long run increase both value provided to customers and profitability of the space industrial sector. This concept paper sets out: (1) the goals for a SUMO standard and how it will benefit the space community; (2) background on spacecraft modularity and existing related standards; (3) the proposed technical scope of the current standardization effort; and (4) an approach for creating a SUMO standard.

  2. Kant e o sumo bem comunitário

    Directory of Open Access Journals (Sweden)

    Letícia Machado Spinelli

    2012-08-01

    Full Text Available http://dx.doi.org/10.5007/1677-2954.2012v11n1p37 O que aqui nos propomos fazer é explicitar o conceito de sumo bem enquanto bem comunitário, em que se evidencia um desdobramento da formulação do conceito de sumo bem sob a perspectiva de um bem coletivo. Esse conceito foi apresentado por Kant na terceira parte de A religião nos limites da simples razão, contexto no qual se dedicou a tratar da noção de um progresso moral nos termos de uma comunidade ética.

  3. The Sumo World through the gambling cases in 2010 The Sumo World as Seen Through the Gambling Cases of 2010

    Directory of Open Access Journals (Sweden)

    Keiko Morita

    2012-11-01

    Full Text Available In summer, 2010, gambling cases of sumo wrestlers and bosses and the connection with the gangsters were reported day after day through mass media. This article chases the summary of a chain of these cases and the correspondence of the Sumo Association for it. The article looks at the present conditions of a unique competition called sumo from these a series of cases. At first, it examines the summary of the illegal gambling case of Ozeki Kotomitsuki, the top of Japanese sumo wrestlers, and the correspondence of Sumo Association and other social organs. It takes up a problem about holding the tournament at Nagoya this summer next. And it relates to dark customs of sumo world that a series of cases are able to happen and tries to explain such customs through the tradition of sumo. While writing this article, the news to ask the self-purification capacity of the Association and to suggest relations with the sumo world and the gangsters are reported every day. The darkness of the sumo world seems to be wide and deep.This article is in Japanese. In summer 2010, a scandal in gambling on baseball matches implicating sumo wrestlers and their bosses was reported day after day by Japanese mass media. This scandal revealed the connections between the sumo world and criminal organizations and lead to unprecedented actions, such as the dismissal of the active Ozeki Kotomitsuki, one of the top Japanese sumo wrestlers, and the suspension of live television broadcasting of the sumo tournament. The reason why the Japanese public instigated mass public debates may relate to the notion that sumo has an intimate association with the gods, reinforced with its special position in Japanese tradition and culture. This article first presents a brief history of sumo and explains its particular characteristics. Then it provides a summary of the chain of events in the baseball gambling scandal and the responses from the Japan Sumo Association. In conjunction with

  4. Binding properties of SUMO-interacting motifs (SIMs) in yeast.

    Science.gov (United States)

    Jardin, Christophe; Horn, Anselm H C; Sticht, Heinrich

    2015-03-01

    Small ubiquitin-like modifier (SUMO) conjugation and interaction play an essential role in many cellular processes. A large number of yeast proteins is known to interact non-covalently with SUMO via short SUMO-interacting motifs (SIMs), but the structural details of this interaction are yet poorly characterized. In the present work, sequence analysis of a large dataset of 148 yeast SIMs revealed the existence of a hydrophobic core binding motif and a preference for acidic residues either within or adjacent to the core motif. Thus the sequence properties of yeast SIMs are highly similar to those described for human. Molecular dynamics simulations were performed to investigate the binding preferences for four representative SIM peptides differing in the number and distribution of acidic residues. Furthermore, the relative stability of two previously observed alternative binding orientations (parallel, antiparallel) was assessed. For all SIMs investigated, the antiparallel binding mode remained stable in the simulations and the SIMs were tightly bound via their hydrophobic core residues supplemented by polar interactions of the acidic residues. In contrary, the stability of the parallel binding mode is more dependent on the sequence features of the SIM motif like the number and position of acidic residues or the presence of additional adjacent interaction motifs. This information should be helpful to enhance the prediction of SIMs and their binding properties in different organisms to facilitate the reconstruction of the SUMO interactome.

  5. Identification and molecular properties of SUMO-binding proteins in arabidopsis

    KAUST Repository

    Park, Hyeongcheol

    2011-05-20

    Reversible conjugation of the small ubiquitin modifier (SUMO) peptide to proteins (SUMOylation) plays important roles in cellular processes in animals and yeasts. However, little is known about plant SUMO targets. To identify SUMO substrates in Arabidopsis and to probe for biological functions of SUMO proteins, we constructed 6xHis-3xFLAG fused AtSUMO1 (HFAtSUMO1) controlled by the CaMV35S promoter for transformation into Arabidopsis Col-0. After heat treatment, an increased sumoylation pattern was detected in the transgenic plants. SUMO1-modified proteins were selected after two-dimensional gel electrophoresis (2-DE) image analysis and identified using matrix-assisted laser-desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). We identified 27 proteins involved in a variety of processes such as nucleic acid metabolism, signaling, metabolism, and including proteins of unknown functions. Binding and sumoylation patterns were confirmed independently. Surprisingly, MCM3 (At5G46280), a DNA replication licensing factor, only interacted with and became sumoylated by AtSUMO1, but not by SUMO1ΔGG or AtSUMO3. The results suggest specific interactions between sumoylation targets and particular sumoylation enzymes. ©2011 KSMCB.

  6. Regulation of SUMO2 Target Proteins by the Proteasome in Human Cells Exposed to Replication Stress

    DEFF Research Database (Denmark)

    Bursomanno, Sara; McGouran, Joanna F; Kessler, Benedikt M

    2015-01-01

    In human cells, SUMO2 is predominantly conjugated to target proteins in response to cellular stress. Previous studies suggested that proteins conjugated to SUMO2, but not to SUMO1, could be regulated by the ubiquitin-mediated proteasome system. Hence, we set out to understand the role of the prot......In human cells, SUMO2 is predominantly conjugated to target proteins in response to cellular stress. Previous studies suggested that proteins conjugated to SUMO2, but not to SUMO1, could be regulated by the ubiquitin-mediated proteasome system. Hence, we set out to understand the role...... of genome instability, which is suggested to drive tumorigenesis and possibly aging, our data will facilitate future functional studies in the fields of DNA metabolism and cancer biology....

  7. Genetic Polymorphism of SUMO-Specific Cysteine Proteases - SENP1 and SENP2 in Breast Cancer.

    Science.gov (United States)

    Mirecka, Alicja; Morawiec, Zbigniew; Wozniak, Katarzyna

    2016-10-01

    SENP proteases take part in post-translational modification of proteins known as sumoylation. They catalyze three distinct processes during sumoylation: processing of SUMO protein, deconjugation of SUMO from the target protein, and chain editing which mentions to the dismantling of SUMO chain. Many proteins that are involved in the basic processes of cells, such as regulation of transcription, DNA repair or cell cycle control, are sumoylated. The aim of these studies was to investigate an association between polymorphic variants (SNPs) of the SENP1 gene (c.1691 + 36C > T, rs12297820) and SENP2 gene (c.902C > A, p.Thr301Lys, rs6762208) and a risk of breast cancer occurrence. We performed a case-control study in 324 breast cancer cases and 335 controls using PCR-RLFP. In the case of the SENP1 gene polymorphism we did not find any association between this polymorphism and breast cancer risk. In the case of SENP2 gene polymorphism we observed higher risk of breast cancer for carriers of the A allele (OR =1.33; 95 % CI 1.04-1.69). Our analysis also showed the genotype C/C (OR =0.67, 95 % CI 0.48-0.93) and the allele C (OR =0.75, 95 % CI 0.59-0.69) of this polymorphism decrease a risk of breast cancer. We also checked the distribution of genotypes and frequency of alleles of the SENP1 and SENP2 genes polymorphisms in groups of patients with different hormone receptor status, patients with positive and negative lymph node status and patients with different tumor grade. Odds ratio analysis showed a higher risk of metastases in women with the genotype C/C (OR =2.07, 95 % CI 1.06-4.05) and allele C (OR =2.10 95 % CI 1.10-4.01) of the c.1691 + 36C > T SENP1 gene polymorphism. Moreover, we observed reduced risk in women with the allele T (OR =0.48, 95 % CI 0.25-0.91) in this polymorphic site. In the case of SENP2 gene polymorphism we observed that the A/A genotype correlated with the lack of estrogen receptor (OR =1.94, 95 % CI 1.04-3.62). Our results suggest

  8. SUMO and SUMO-Conjugating Enzyme E2 UBC9 Are Involved in White Spot Syndrome Virus Infection in Fenneropenaeus chinensis.

    Directory of Open Access Journals (Sweden)

    Xiaoqian Tang

    Full Text Available In previous work, small ubiquitin-like modifier (SUMO in hemocytes of Chinese shrimp Fenneropenaeus chinensis was found to be up-regulated post-white spot syndrome virus (WSSV infection using proteomic approach. However, the role of SUMO in viral infection is still unclear. In the present work, full length cDNAs of SUMO (FcSUMO and SUMO-conjugating enzyme E2 UBC9 (FcUBC9 were cloned from F. chinensis using rapid amplification of cDNA ends approach. The open reading frame (ORF of FcSUMO encoded a 93 amino acids peptide with the predicted molecular weight (M.W of 10.55 kDa, and the UBC9 ORF encoded a 160 amino acids peptide with the predicted M.W of 18.35 kDa. By quantitative real-time RT-PCR, higher mRNA transcription levels of FcSUMO and FcUBC9 were detected in hemocytes and ovary of F. chinensis, and the two genes were significantly up-regulated post WSSV infection. Subsequently, the recombinant proteins of FcSUMO and FcUBC9 were expressed in Escherichia coli BL21 (DE3, and employed as immunogens for the production of polyclonal antibody (PAb. Indirect immunofluorescence assay revealed that the FcSUMO and UBC9 proteins were mainly located in the hemocytes nuclei. By western blotting, a 13.5 kDa protein and a 18.7 kDa protein in hemocytes were recognized by the PAb against SUMO or UBC9 respectively. Furthermore, gene silencing of FcSUMO and FcUBC9 were performed using RNA interference, and the results showed that the number of WSSV copies and the viral gene expressions were inhibited by knockdown of either SUMO or UBC9, and the mortalities of shrimp were also reduced. These results indicated that FcSUMO and FcUBC9 played important roles in WSSV infection.

  9. Construction of fusion expression vector pET22b-SUMO-FGFR4 and optimization of expression conditions in E.coli%融合表达载体 pET22b-SUMO-FGFR4的构建及其在大肠杆菌中表达条件的优化

    Institute of Scientific and Technical Information of China (English)

    刘微; 姚杨; 马萧萧; 邓裕宣; 梅迪; 刘磊; 王会岩

    2016-01-01

    目的:设计合成小泛素修饰物-成纤维细胞生长因子受体4(SUMO-FGFR4)基因,构建 pET22b-SUMO-FGFR4表达载体,并对其表达条件进行优化。方法:采用 Overlap PCR 方法制备 SUMO-FGFR4融合基因,并连接到原核表达载体 pET22b 中,获得 pET22b-SUMO-FGFR4重组表达载体。以乳糖为诱导剂,观察乳糖浓度、诱导时机、诱导温度、诱导时间和乳糖的添加方式等因素对 SUMO-FGFR4蛋白表达量的影响,确定最佳诱导条件,并进行重组蛋白的可溶性分析。结果:pET22b-SUMO-FGFR4表达的融合蛋白在相对分子质量40000处显示目标条带,并与 FGFR4抗体特异性结合。融合蛋白在乳糖终浓度为1.0 g·L-1、诱导时间为3 h、诱导时机 A (600)值为0.8、诱导温度为37℃时表达量最高,乳糖的添加方式对 SUMO-FGFR4融合蛋白的表达量无明显影响。乳糖作为诱导剂比传统诱导剂 IPTG 诱导 SUMO-FGFR4融合蛋白的表达量高7.5%,融合蛋白以包涵体形式为主。结论:以乳糖作为诱导剂,成功表达了 SUMO-FGFR4融合蛋白,确定了融合蛋白的最佳表达条件。%Objective:To design the small ubiquitin modification-fibroblast growth factor receptor 4 (SUMO-FGFR4) fusion gene and construct the expression vector pET22b-SUMO-FGFR4, to optimize the expression conditions. Methods:The SUMO-FGFR4 fusion gene was obtained by Overlap PCR and was connected to pET22b;the recombinant expression vector pET22b-SUMO-FGFR4 was obtained. The influence of lactose concentration, induction time,induction temperature,induction point and adding mode of lactose in the expression levels was observed,and the best induction condition was determined; then the solubility of recombinant protein was analyzed.Results:The SUMO-FGFR4 fusion protein was highly expressed,the molecular weight of the fusion protein was about 40 000 and it could bind with FGFR4 specific antibody.When the lactose concentration

  10. Whole-genome duplications followed by tandem duplications drive diversification of the protein modifier SUMO in Angiosperms

    NARCIS (Netherlands)

    Hammoudi, V.; Vlachakis, G.; Schranz, M.E.; van den Burg, H.A.

    2016-01-01

    The ubiquitin-like modifier (UBL) SUMO (Small Ubiquitin-Like Modifier) regulates protein function. Structural rather than sequence homology typifies UBL families. However, individual UBL types, such as SUMO, show remarkable sequence conservation. Selection pressure also operates at the SUMO gene cop

  11. Whole-genome duplications followed by tandem duplications drive diversification of the protein modifier SUMO in Angiosperms

    NARCIS (Netherlands)

    Hammoudi, Valentin; Vlachakis, Georgios; Schranz, Eric; Burg, van den Harrold A.

    2016-01-01

    The ubiquitin-like modifier (UBL) SUMO (Small Ubiquitin-Like Modifier) regulates protein function. Structural rather than sequence homology typifies UBL families. However, individual UBL types, such as SUMO, show remarkable sequence conservation. Selection pressure also operates at the SUMO gene

  12. Whole-genome duplications followed by tandem duplications drive diversification of the protein modifier SUMO in Angiosperms

    NARCIS (Netherlands)

    Hammoudi, V.; Vlachakis, G.; Schranz, M.E.; van den Burg, H.A.

    2016-01-01

    The ubiquitin-like modifier (UBL) SUMO (Small Ubiquitin-Like Modifier) regulates protein function. Structural rather than sequence homology typifies UBL families. However, individual UBL types, such as SUMO, show remarkable sequence conservation. Selection pressure also operates at the SUMO gene cop

  13. Whole-genome duplications followed by tandem duplications drive diversification of the protein modifier SUMO in Angiosperms

    NARCIS (Netherlands)

    Hammoudi, Valentin; Vlachakis, Georgios; Schranz, Eric; Burg, van den Harrold A.

    2016-01-01

    The ubiquitin-like modifier (UBL) SUMO (Small Ubiquitin-Like Modifier) regulates protein function. Structural rather than sequence homology typifies UBL families. However, individual UBL types, such as SUMO, show remarkable sequence conservation. Selection pressure also operates at the SUMO gene

  14. Characterizing the N- and C-terminal Small Ubiquitin-like Modifier (SUMO)-interacting Motifs of the Scaffold Protein DAXX

    NARCIS (Netherlands)

    Escobar-Cabrera, E.; Okon, M.; Lau, D.K.W.; Dart, C.F.; Bonvin, A.M.J.J.; McIntosh, L.P.

    2011-01-01

    DAXX is a scaffold protein with diverse roles that often depend upon binding SUMO via its N- and/or C-terminal SUMO-interacting motifs (SIM-N and SIM-C). Using NMR spectroscopy, we characterized the in vitro binding properties of peptide models of SIM-N and SIM-C to SUMO-1 and SUMO-2. In each case,

  15. The nucleolar SUMO-specific protease SMT3IP1/SENP3 attenuates Mdm2-mediated p53 ubiquitination and degradation

    Energy Technology Data Exchange (ETDEWEB)

    Nishida, Tamotsu, E-mail: nishida@gene.mie-u.ac.jp [Department of Human Functional Genomics, Life Science Research Center, Mie University, 1577 Kurima-machiya, Tsu 514-8507 (Japan); Yamada, Yoshiji [Department of Human Functional Genomics, Life Science Research Center, Mie University, 1577 Kurima-machiya, Tsu 514-8507 (Japan)

    2011-03-11

    Research highlights: {yields} SMT3IP1 interacts with p53 and Mdm2, and desumoylates both proteins. {yields} SMT3IP1 competes with p53 for binding to the central acidic domain of Mdm2. {yields} SMT3IP1 binding to Mdm2 inhibits Mdm2-mediated p53 ubiquitination and degradation. {yields} We postulate that SMT3IP1 acts as a new regulator of the p53-Mdm2 pathway. -- Abstract: SUMO (small ubiquitin-like modifier) modification plays multiple roles in several cellular processes. Sumoylation is reversibly regulated by SUMO-specific proteases. SUMO-specific proteases have recently been implicated in cell proliferation and early embryogenesis, but the underlying mechanisms remain unknown. Here, we show that a nucleolar SUMO-specific protease, SMT3IP1/SENP3, controls the p53-Mdm2 pathway. We found that SMT3IP1 interacts with p53 and Mdm2, and desumoylates both proteins. Overexpression of SMT3IP1 in cells resulted in the accumulation of Mdm2 in the nucleolus and increased stability of the p53 protein. In addition, SMT3IP1 bound to the acidic domain of Mdm2, which also mediates the p53 interaction, and competed with p53 for binding. Increasing expression of SMT3IP1 suppressed Mdm2-mediated p53 ubiquitination and subsequent proteasomal degradation. Interestingly, the desumoylation activity of SMT3IP1 was not necessary for p53 stabilization. These results suggest that SMT3IP1 is a new regulator of the p53-Mdm2 pathway.

  16. SUMO-1 gene transfer improves cardiac function in a large-animal model of heart failure.

    Science.gov (United States)

    Tilemann, Lisa; Lee, Ahyoung; Ishikawa, Kiyotake; Aguero, Jaume; Rapti, Kleopatra; Santos-Gallego, Carlos; Kohlbrenner, Erik; Fish, Kenneth M; Kho, Changwon; Hajjar, Roger J

    2013-11-13

    Recently, the impact of small ubiquitin-related modifier 1 (SUMO-1) on the regulation and preservation of sarcoplasmic reticulum calcium adenosine triphosphatase (SERCA2a) function was discovered. The amount of myocardial SUMO-1 is decreased in failing hearts, and its knockdown results in severe heart failure (HF) in mice. In a previous study, we showed that SUMO-1 gene transfer substantially improved cardiac function in a murine model of pressure overload-induced HF. Toward clinical translation, we evaluated in this study the effects of SUMO-1 gene transfer in a swine model of ischemic HF. One month after balloon occlusion of the proximal left anterior descending artery followed by reperfusion, the animals were randomized to receive either SUMO-1 at two doses, SERCA2a, or both by adeno-associated vector type 1 (AAV1) gene transfer via antegrade coronary infusion. Control animals received saline infusions. After gene delivery, there was a significant increase in the maximum rate of pressure rise [dP/dt(max)] that was most pronounced in the group that received both SUMO-1 and SERCA2a. The left ventricular ejection fraction (LVEF) improved after high-dose SUMO-1 with or without SERCA2a gene delivery, whereas there was a decline in LVEF in the animals receiving saline. Furthermore, the dilatation of LV volumes was prevented in the treatment groups. SUMO-1 gene transfer therefore improved cardiac function and stabilized LV volumes in a large-animal model of HF. These results support the critical role of SUMO-1 in SERCA2a function and underline the therapeutic potential of SUMO-1 for HF patients.

  17. SUMO-interacting motifs of human TRIM5α are important for antiviral activity.

    Directory of Open Access Journals (Sweden)

    Gloria Arriagada

    2011-04-01

    Full Text Available Human TRIM5α potently restricts particular strains of murine leukemia viruses (the so-called N-tropic strains but not others (the B- or NB-tropic strains during early stages of infection. We show that overexpression of SUMO-1 in human 293T cells, but not in mouse MDTF cells, profoundly blocks N-MLV infection. This block is dependent on the tropism of the incoming virus, as neither B-, NB-, nor the mutant R110E of N-MLV CA (a B-tropic switch are affected by SUMO-1 overexpression. The block occurred prior to reverse transcription and could be abrogated by large amounts of restricted virus. Knockdown of TRIM5α in 293T SUMO-1-overexpressing cells resulted in ablation of the SUMO-1 antiviral effects, and this loss of restriction could be restored by expression of a human TRIM5α shRNA-resistant plasmid. Amino acid sequence analysis of human TRIM5α revealed a consensus SUMO conjugation site at the N-terminus and three putative SUMO interacting motifs (SIMs in the B30.2 domain. Mutations of the TRIM5α consensus SUMO conjugation site did not affect the antiviral activity of TRIM5α in any of the cell types tested. Mutation of the SIM consensus sequences, however, abolished TRIM5α antiviral activity against N-MLV. Mutation of lysines at a potential site of SUMOylation in the CA region of the Gag gene reduced the SUMO-1 block and the TRIM5α restriction of N-MLV. Our data suggest a novel aspect of TRIM5α-mediated restriction, in which the presence of intact SIMs in TRIM5α, and also the SUMO conjugation of CA, are required for restriction. We propose that at least a portion of the antiviral activity of TRIM5α is mediated through the binding of its SIMs to SUMO-conjugated CA.

  18. The p150 subunit of CAF-1 causes association of SUMO2/3 with the DNA replication foci

    Energy Technology Data Exchange (ETDEWEB)

    Uwada, Junsuke [Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Global COE (Centers of Excellence) Program, Global Initiative Center for Pulsed Power Engineering, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Tanaka, Niina; Yamaguchi, Yutaro; Uchimura, Yasuhiro [Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Shibahara, Kei-ichi [Department of Integrated Genetics, National Institute of Genetics, Research Organization of Information and Systems, Mishima (Japan); Nakao, Mitsuyoshi [Department of Medical Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto (Japan); Saitoh, Hisato, E-mail: hisa@kumamoto-u.ac.jp [Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Global COE (Centers of Excellence) Program, Global Initiative Center for Pulsed Power Engineering, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan)

    2010-01-01

    The small ubiquitin-related modifier 2/3 (SUMO2/3) can be post-translationally conjugated to a wide variety of proteins constituting chromatin, the platform for genetic and epigenetic regulation. Nevertheless, it is unclear how SUMO2/3 and SUMO2/3-modified proteins are delivered to the chromatin fibers. Here we report that the largest subunit of chromatin assembly factor 1 (CAF-1), human p150, interacts directly and preferentially with SUMO2/3. Amino acid residue of 98-105 in p150 is essential and sufficient for SUMO2/3 interaction. p150-SUMO2/3 interaction coincided with regions that replicate chromatin fibers, because accumulation of the proliferating cell nuclear antigen (PCNA), and incorporation of bromodeoxyuridine (BrdU) were detected at foci co-localized with both p150 and SUMO2/3 during the S-phase in a cell line expressing epitope-tagged p150. Although inhibition of SUMO2/3 expression had only a small effect on p150 deposition on the replication sites, depletion of p150 led to delocalization of SUMO2/3 from the replication foci. Furthermore, p150 mutants deficient in SUMO2/3 interaction, caused a major reduction of SUMO2/3 at the replication foci. Thus, our findings suggest an expanded role of p150 as a SUMO2/3-interacting factor, and raise the intriguing possibility that p150 plays a role in promoting delivery of SUMO2/3 or SUMO2/3-modified proteins (or both) on chromatin fibers during replication.

  19. A high-yield double-purification proteomics strategy for the identification of SUMO sites.

    Science.gov (United States)

    Hendriks, Ivo A; Vertegaal, Alfred C O

    2016-09-01

    The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its involvement in diseases such as cancer. Here we describe a protocol for the lysine-deficient (K0) method for efficient identification of SUMOylated lysines by mass spectrometry (MS). To our knowledge, the K0 method is the only currently available method that can routinely identify >1,000 SUMO sites in mammalian cells under standard growth conditions. The K0 strategy relies on introducing a His10-tagged SUMO wherein all lysines have been substituted to arginines. Lysine deficiency renders the SUMO immune to digestion by the endoproteinase Lys-C, which in turn allows for stringent and high-yield tandem purification through the His10 tag. In addition, the His10-tagged SUMO also contains a C-terminal Q87R mutation, which accommodates generation of SUMO-site peptides with a QQTGG mass remnant after digestion with trypsin. This remnant possesses a unique mass signature and readily generates diagnostic ions in the fragment ion scans, which increases SUMO-site identification confidence. The K0 method can be applied in any mammalian cell line or in any model system that allows for integration of the K0-SUMO construct. From the moment of cell lysis, the K0 method takes ∼7 d to perform.

  20. Identifications of SUMO-1 cDNA and Its Expression Patterns in Pacific White Shrimp Litopeanaeus vannamei

    Directory of Open Access Journals (Sweden)

    Yanisa Laoong-u-thai, Baoping Zhao, Amornrat Phongdara, Harry Ako, Jinzeng Yang

    2009-01-01

    Full Text Available Small ubiquitin-like modifiers (SUMO work in a similar way as ubiquitin to alter the biological properties of a target protein by conjugation. A shrimp SUMO cDNA named LvSUMO-1 was identified in Litopenaeus vannamei. LvSUMO-1 cDNA contains a coding sequence of 282 nucleotides with untranslated regions of 37 bp at 5'-end and 347 bp at 3'-end, respectively. The deduced 93 amino acids exhibit 83% identity with the Western Honeybee SUMO-1, and more than 65% homologies with human and mouse SUMO-1. LvSUMO-1 mRNA is expressed in most L. vannamei tissues with the highest level in hepatopancrease. The mRNA expression of LvSUMO-1 over development stages in L. Vammamei is distinguished by a low level in nauplius stage and relatively high level in postlarva stage with continuous expression until juvenile stage. The LvSUMO-1 protein and its conjugated proteins are detected in both cytoplasm and nucleus in several tissues. Interestingly, LvSUMO-1 mRNA levels are high in abdominal muscle during the premolt stage, wherein it has significant activities of protein degradation, suggesting its possible role in the regulation of shrimp muscle protein degradation.

  1. Complex SUMO-1 regulation of cardiac transcription factor Nkx2-5.

    Directory of Open Access Journals (Sweden)

    Mauro W Costa

    Full Text Available Reversible post-translational protein modifications such as SUMOylation add complexity to cardiac transcriptional regulation. The homeodomain transcription factor Nkx2-5/Csx is essential for heart specification and morphogenesis. It has been previously suggested that SUMOylation of lysine 51 (K51 of Nkx2-5 is essential for its DNA binding and transcriptional activation. Here, we confirm that SUMOylation strongly enhances Nkx2-5 transcriptional activity and that residue K51 of Nkx2-5 is a SUMOylation target. However, in a range of cultured cell lines we find that a point mutation of K51 to arginine (K51R does not affect Nkx2-5 activity or DNA binding, suggesting the existence of additional Nkx2-5 SUMOylated residues. Using biochemical assays, we demonstrate that Nkx2-5 is SUMOylated on at least one additional site, and this is the predominant site in cardiac cells. The second site is either non-canonical or a "shifting" site, as mutation of predicted consensus sites and indeed every individual lysine in the context of the K51R mutation failed to impair Nkx2-5 transcriptional synergism with SUMO, or its nuclear localization and DNA binding. We also observe SUMOylation of Nkx2-5 cofactors, which may be critical to Nkx2-5 regulation. Our data reveal highly complex regulatory mechanisms driven by SUMOylation to modulate Nkx2-5 activity.

  2. The SUMO Ship Detector Algorithm for Satellite Radar Images

    Directory of Open Access Journals (Sweden)

    Harm Greidanus

    2017-03-01

    Full Text Available Search for Unidentified Maritime Objects (SUMO is an algorithm for ship detection in satellite Synthetic Aperture Radar (SAR images. It has been developed over the course of more than 15 years, using a large amount of SAR images from almost all available SAR satellites operating in L-, C- and X-band. As validated by benchmark tests, it performs very well on a wide range of SAR image modes (from Spotlight to ScanSAR and resolutions (from 1–100 m and for all types and sizes of ships, within the physical limits imposed by the radar imaging. This paper describes, in detail, the algorithmic approach in all of the steps of the ship detection: land masking, clutter estimation, detection thresholding, target clustering, ship attribute estimation and false alarm suppression. SUMO is a pixel-based CFAR (Constant False Alarm Rate detector for multi-look radar images. It assumes a K distribution for the sea clutter, corrected however for deviations of the actual sea clutter from this distribution, implementing a fast and robust method for the clutter background estimation. The clustering of detected pixels into targets (ships uses several thresholds to deal with the typically irregular distribution of the radar backscatter over a ship. In a multi-polarization image, the different channels are fused. Azimuth ambiguities, a common source of false alarms in ship detection, are removed. A reliability indicator is computed for each target. In post-processing, using the results of a series of images, additional false alarms from recurrent (fixed targets including range ambiguities are also removed. SUMO can run in semi-automatic mode, where an operator can verify each detected target. It can also run in fully automatic mode, where batches of over 10,000 images have successfully been processed in less than two hours. The number of satellite SAR systems keeps increasing, as does their application to maritime surveillance. The open data policy of the EU

  3. SUMO/FREND: vision system for autonomous satellite grapple

    Science.gov (United States)

    Obermark, Jerome; Creamer, Glenn; Kelm, Bernard E.; Wagner, William; Henshaw, C. Glen

    2007-04-01

    SUMO/FREND is a risk reduction program for an advanced servicing spacecraft sponsored by DARPA and executed by the Naval Center for Space Technology at the Naval Research Laboratory in Washington, DC. The overall program will demonstrate the integration of many techniques needed in order to autonomously rendezvous and capture customer satellites at geosynchronous orbits. A flight-qualifiable payload is currently under development to prove out challenging aspects of the mission. The grappling process presents computer vision challenges to properly identify and guide the final step in joining the pursuer craft to the customer. This paper will provide an overview of the current status of the project with an emphasis on the challenges, techniques, and directions of the machine vision processes to guide the grappling.

  4. VP6-SUMO Self-Assembly as Nanocarriers for Gastrointestinal Delivery

    Directory of Open Access Journals (Sweden)

    V. Palmieri

    2015-01-01

    Full Text Available High proteolytic degradation and poor absorption through epithelial barriers are major challenges to successful oral delivery of therapeutics. Nanoparticle platforms can enhance drug stability and extend the residence time in gastrointestinal (GI tract. However, drug delivery systems are often inactivated in acidic environment of stomach or suffer poor absorption from intestinal cells due to the mucus layer. To overcome these issues we developed a drug delivery system constituted by a protein construct made by a Rotavirus capsid protein (VP6 and the small ubiquitin-like modifier SUMO. This chimeric construct allows specificity towards intestinal cells, the Rotavirus natural target, combined by an enhanced stability given by the eukaryotic protein transporter SUMO. Furthermore SUMO can act as a molecular switch that facilitates import/export of its ligand to the nucleus, the hypersensitive subcellular site target of many cell killing therapies. In this paper we show that SUMO-VP6 constructs self-assembly into stable nanocarriers. SUMO-VP6 nanocarriers display ideal features for drug delivery: a small size and high monodispersity, a high stability in different pH conditions and a high uptake in the nuclear and cytoplasmic compartment of intestinal cells. These features make SUMO-VP6 nanocarriers a promising novel system for oral delivery of poorly soluble drugs.

  5. SUMO: A small unmanned meteorological observer for atmospheric boundary layer research

    Energy Technology Data Exchange (ETDEWEB)

    Reuder, J; Jonassen, M; Mayer, S [Geophysical Institute, University of Bergen, Allegaten 70, 5009 Bergen (Norway); Brisset, P [Ecole Nationale de l' Aviation Civile (ENAC), 7 avenue Edouard Belin, 31055 Toulouse (France); Mueller, M [Orleansstrasse 26a, 31135 Hildesheim (Germany)], E-mail: joachim.reuder@gfi.uib.no, E-mail: pascal.brisset@enac.fr, E-mail: marius.jonassen@gfi.uib.no, E-mail: martin@pfump.org, E-mail: stephanie.mayer@gfi.uib.no

    2008-05-01

    A new system for atmospheric measurements in the lower troposphere has been developed and successfully tested. The presented Small Unmanned Meteorological Observer (SUMO) is based on a light-weighted commercially available model airplane, equipped with an autopilot and meteorological sensors for temperature, humidity and pressure. During the 5 week field campaign FLOHOF (Flow over and around HofsjoUkull) in Central Iceland the system has been successfully tested in July/August 2007. Atmospheric profiles of temperature, humidity, wind speed and wind direction have been determined up to 3500 m above ground. In addition the applicability of SUMO for horizontal surveys up to 4 km away from the launch site has been approved. During a 3 week campaign on and around Spitsbergen in February/March 2008 the SUMO system also proved its functionality under harsh polar conditions, reaching altitudes above 1500 m at ground temperatures of -20 deg. C and wind speeds up to 15 m s{sup -1}. With its wingspan of 80 cm, its length of 75 cm and its weight of below 600 g, SUMO is easy to transport and operate even in remote areas. The direct material costs for one SUMO unit, including airplane, autopilot and sensors are below 1200 Euro. Assuming at least several tenths of flights for each airframe, SUMO provides a cost-efficient measurement system with a large potential to close the existing observational gap of reasonable atmospheric measurement systems in between meteorological masts/towers and radiosondes.

  6. SUMO-1 regulates the conformational dynamics of Thymine-DNA Glycosylase regulatory domain and competes with its DNA binding activity

    Directory of Open Access Journals (Sweden)

    Eilebrecht Sebastian

    2011-02-01

    Full Text Available Abstract Background The human thymine-DNA glycosylase (TDG plays a dual role in base excision repair of G:U/T mismatches and in transcription. Regulation of TDG activity by SUMO-1 conjugation was shown to act on both functions. Furthermore, TDG can interact with SUMO-1 in a non-covalent manner. Results Using NMR spectroscopy we have determined distinct conformational changes in TDG upon either covalent sumoylation on lysine 330 or intermolecular SUMO-1 binding through a unique SUMO-binding motif (SBM localized in the C-terminal region of TDG. The non-covalent SUMO-1 binding induces a conformational change of the TDG amino-terminal regulatory domain (RD. Such conformational dynamics do not exist with covalent SUMO-1 attachment and could potentially play a broader role in the regulation of TDG functions for instance during transcription. Both covalent and non-covalent processes activate TDG G:U repair similarly. Surprisingly, despite a dissociation of the SBM/SUMO-1 complex in presence of a DNA substrate, SUMO-1 preserves its ability to stimulate TDG activity indicating that the non-covalent interactions are not directly involved in the regulation of TDG activity. SUMO-1 instead acts, as demonstrated here, indirectly by competing with the regulatory domain of TDG for DNA binding. Conclusions SUMO-1 increases the enzymatic turnover of TDG by overcoming the product-inhibition of TDG on apurinic sites. The mechanism involves a competitive DNA binding activity of SUMO-1 towards the regulatory domain of TDG. This mechanism might be a general feature of SUMO-1 regulation of other DNA-bound factors such as transcription regulatory proteins.

  7. Association of SUMO4 M55V polymorphism with autoimmune diabetes in Latvian patients.

    Science.gov (United States)

    Sedimbi, Saikiran K; Shastry, Arun; Park, Yongsoo; Rumba, Ingrida; Sanjeevi, Carani B

    2006-10-01

    Small ubiquitin-related modifier (SUMO4), located in IDDM5, has been identified as a potential susceptibility gene for type 1 diabetes mellitus (T1DM). The novel polymorphism M55V, causing an amino acid change in the evolutionarily conserved met55 residue has been shown to activate the nuclear factor kappaB (NF-kappaB), hence the suspected role of SUMO4 in the pathogenicity of T1DM. The M55V polymorphism has been shown to be associated with susceptibility to T1DM in Asians, but not in Caucasians. Latent autoimmune diabetes in adults (LADA) is a slowly progressive form of T1DM and SUMO4 M55V has not been studied in LADA to date. The current study aims to test whether Latvians are similar to Caucasians in susceptibility to autoimmune diabetes (T1DM and LADA), with respect to SUMO4 M55V. We studied, age- and sex-matched, Latvian T1DM patients (n = 100) and healthy controls (n = 90) and LADA patients (n = 45) and healthy controls (n = 95). SUMO4 M55V polymorphism was analyzed using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). The allelic frequencies of the A and G alleles were compared with HLA DR3-DR4-positive and HLA DR3-DR4-negative patients to identify any potential relation between HLA DR3-DR4 and SUMO4 M55V. We found no significant association between SUMO4 M55V and T1DM susceptibility in Latvians, the results being in concurrence with the previous studies in Caucasians of British and Canadian origin. Comparison of the A and G alleles with HLA DR3-DR4 did not result in any significant P values. No significant association was found between SUMO4 M55V and LADA. SUMO4 M55V is not associated with susceptibility to T1DM and LADA in Latvians, and Latvians exhibit similarity to other Caucasians with respect to association of SUMO4 M55V with autoimmune diabetes.

  8. The SUMO protease SENP1 is required for cohesion maintenance and mitotic arrest following spindle poison treatment

    Energy Technology Data Exchange (ETDEWEB)

    Era, Saho [Fondazione IFOM, Istituto FIRC di Oncologia Molecolare, IFOM-IEO campus, Via Adamello 16, 20139 Milan (Italy); Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501 (Japan); Abe, Takuya; Arakawa, Hiroshi [Fondazione IFOM, Istituto FIRC di Oncologia Molecolare, IFOM-IEO campus, Via Adamello 16, 20139 Milan (Italy); Kobayashi, Shunsuke [Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501 (Japan); Szakal, Barnabas [Fondazione IFOM, Istituto FIRC di Oncologia Molecolare, IFOM-IEO campus, Via Adamello 16, 20139 Milan (Italy); Yoshikawa, Yusuke; Motegi, Akira; Takeda, Shunichi [Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshida-Konoe, Sakyo-ku, Kyoto 606-8501 (Japan); Branzei, Dana, E-mail: dana.branzei@ifom.eu [Fondazione IFOM, Istituto FIRC di Oncologia Molecolare, IFOM-IEO campus, Via Adamello 16, 20139 Milan (Italy)

    2012-09-28

    Highlights: Black-Right-Pointing-Pointer SENP1 knockout chicken DT40 cells are hypersensitive to spindle poisons. Black-Right-Pointing-Pointer Spindle poison treatment of SENP1{sup -/-} cells leads to increased mitotic slippage. Black-Right-Pointing-Pointer Mitotic slippage in SENP1{sup -/-} cells associates with apoptosis and endoreplication. Black-Right-Pointing-Pointer SENP1 counteracts sister chromatid separation during mitotic arrest. Black-Right-Pointing-Pointer Plk1-mediated cohesion down-regulation is involved in colcemid cytotoxicity. -- Abstract: SUMO conjugation is a reversible posttranslational modification that regulates protein function. SENP1 is one of the six SUMO-specific proteases present in vertebrate cells and its altered expression is observed in several carcinomas. To characterize SENP1 role in genome integrity, we generated Senp1 knockout chicken DT40 cells. SENP1{sup -/-} cells show normal proliferation, but are sensitive to spindle poisons. This hypersensitivity correlates with increased sister chromatid separation, mitotic slippage, and apoptosis. To test whether the cohesion defect had a causal relationship with the observed mitotic events, we restored the cohesive status of sister chromatids by introducing the TOP2{alpha}{sup +/-} mutation, which leads to increased catenation, or by inhibiting Plk1 and Aurora B kinases that promote cohesin release from chromosomes during prolonged mitotic arrest. Although TOP2{alpha} is SUMOylated during mitosis, the TOP2{alpha}{sup +/-} mutation had no obvious effect. By contrast, inhibition of Plk1 or Aurora B rescued the hypersensitivity of SENP1{sup -/-} cells to colcemid. In conclusion, we identify SENP1 as a novel factor required for mitotic arrest and cohesion maintenance during prolonged mitotic arrest induced by spindle poisons.

  9. Determination of posttranslational modifications of photoreceptor differentiation factor NRL: focus on SUMOylation.

    Science.gov (United States)

    Roger, Jerome E; Nellissery, Jacob; Swaroop, Anand

    2012-01-01

    Conjugation of SUMO (small ubiquitin-related modifier 1) is a critical posttranslational modification, with significant impact on protein function/activity. Here, we describe direct SUMOylation of GST (glutathione S-transferase)-fusion protein and immunoprecipitation assays for investigating SUMOylation of any protein of interest. We have employed these methods to examine SUMOylation of the basic-motif leucine zipper transcription factor NRL.

  10. Crystal Structure of UBA2[superscript ufd]-Ubc9: Insights into E1-E2 Interactions in Sumo Pathways

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jing; Taherbhoy, Asad M.; Hunt, Harold W.; Seyedin, Steven N.; Miller, David W.; Miller, Darcie J.; Huang, Danny T.; Schulman, Brenda A. (SJCH)

    2012-04-30

    Canonical ubiquitin-like proteins (UBLs) such as ubiquitin, Sumo, NEDD8, and ISG15 are ligated to targets by E1-E2-E3 multienzyme cascades. The Sumo cascade, conserved among all eukaryotes, regulates numerous biological processes including protein localization, transcription, DNA replication, and mitosis. Sumo conjugation is initiated by the heterodimeric Aos1-Uba2 E1 enzyme (in humans called Sae1-Uba2), which activates Sumo's C-terminus, binds the dedicated E2 enzyme Ubc9, and promotes Sumo C-terminal transfer between the Uba2 and Ubc9 catalytic cysteines. To gain insights into details of E1-E2 interactions in the Sumo pathway, we determined crystal structures of the C-terminal ubiquitin fold domain (ufd) from yeast Uba2 (Uba2{sup ufd}), alone and in complex with Ubc9. The overall structures of both yeast Uba2{sup ufd} and Ubc9 superimpose well on their individual human counterparts, suggesting conservation of fundamental features of Sumo conjugation. Docking the Uba2{sup ufd}-Ubc9 and prior full-length human Uba2 structures allows generation of models for steps in Sumo transfer from Uba2 to Ubc9, and supports the notion that Uba2 undergoes remarkable conformational changes during the reaction. Comparisons to previous structures from the NEDD8 cascade demonstrate that UBL cascades generally utilize some parallel E1-E2 interaction surfaces. In addition, the structure of the Uba2{sup ufd}-Ubc9 complex reveals interactions unique to Sumo E1 and E2. Comparison with a previous Ubc9-E3 complex structure demonstrates overlap between Uba2 and E3 binding sites on Ubc9, indicating that loading with Sumo and E3-catalyzed transfer to substrates are strictly separate steps. The results suggest mechanisms establishing specificity and order in Sumo conjugation cascades.

  11. Global analysis of SUMO chain function reveals multiple roles in chromatin regulation.

    Science.gov (United States)

    Srikumar, Tharan; Lewicki, Megan C; Costanzo, Michael; Tkach, Johnny M; van Bakel, Harm; Tsui, Kyle; Johnson, Erica S; Brown, Grant W; Andrews, Brenda J; Boone, Charles; Giaever, Guri; Nislow, Corey; Raught, Brian

    2013-04-01

    Like ubiquitin, the small ubiquitin-related modifier (SUMO) proteins can form oligomeric "chains," but the biological functions of these superstructures are not well understood. Here, we created mutant yeast strains unable to synthesize SUMO chains (smt3(allR)) and subjected them to high-content microscopic screening, synthetic genetic array (SGA) analysis, and high-density transcript profiling to perform the first global analysis of SUMO chain function. This comprehensive assessment identified 144 proteins with altered localization or intensity in smt3(allR) cells, 149 synthetic genetic interactions, and 225 mRNA transcripts (primarily consisting of stress- and nutrient-response genes) that displayed a >1.5-fold increase in expression levels. This information-rich resource strongly implicates SUMO chains in the regulation of chromatin. Indeed, using several different approaches, we demonstrate that SUMO chains are required for the maintenance of normal higher-order chromatin structure and transcriptional repression of environmental stress response genes in budding yeast.

  12. Direct and/or Indirect Roles for SUMO in Modulating Alpha-Synuclein Toxicity

    Directory of Open Access Journals (Sweden)

    Shamini Vijayakumaran

    2015-07-01

    Full Text Available α-Synuclein inclusion bodies are a pathological hallmark of several neurodegenerative diseases, including Parkinson’s disease, and contain aggregated α-synuclein and a variety of recruited factors, including protein chaperones, proteasome components, ubiquitin and the small ubiquitin-like modifier, SUMO-1. Cell culture and animal model studies suggest that misfolded, aggregated α-synuclein is actively translocated via the cytoskeletal system to a region of the cell where other factors that help to lessen the toxic effects can also be recruited. SUMO-1 covalently conjugates to various intracellular target proteins in a way analogous to ubiquitination to alter cellular distribution, function and metabolism and also plays an important role in a growing list of cellular pathways, including exosome secretion and apoptosis. Furthermore, SUMO-1 modified proteins have recently been linked to cell stress responses, such as oxidative stress response and heat shock response, with increased SUMOylation being neuroprotective in some cases. Several recent studies have linked SUMOylation to the ubiquitin-proteasome system, while other evidence implicates the lysosomal pathway. Other reports depict a direct mechanism whereby sumoylation reduced the aggregation tendency of α-synuclein, and reduced the toxicity. However, the precise role of SUMO-1 in neurodegeneration remains unclear. In this review, we explore the potential direct or indirect role(s of SUMO-1 in the cellular response to misfolded α-synuclein in neurodegenerative disorders.

  13. Enhanced protein expression in the baculovirus/insect cell system using engineered SUMO fusions.

    Science.gov (United States)

    Liu, Li; Spurrier, Joshua; Butt, Tauseef R; Strickler, James E

    2008-11-01

    Recombinant protein expression in insect cells varies greatly from protein to protein. A fusion tag that is not only a tool for detection and purification, but also enhances expression and/or solubility would greatly facilitate both structure/function studies and therapeutic protein production. We have shown that fusion of SUMO (small ubiquitin-related modifier) to several test proteins leads to enhanced expression levels in Escherichia coli. In eukaryotic expression systems, however, the SUMO tag could be cleaved by endogenous desumoylase. In order to adapt SUMO-fusion technology to these systems, we have developed an alternative SUMO-derived tag, designated SUMOstar, which is not processed by native SUMO proteases. In the present study, we tested the SUMOstar tag in a baculovirus/insect cell system with several proteins, i.e. mouse UBP43, human tryptase beta II, USP4, USP15, and GFP. Our results demonstrate that fusion to SUMOstar enhanced protein expression levels at least 4-fold compared to either the native or His(6)-tagged proteins. We isolated active SUMOstar tagged UBP43, USP4, USP15, and GFP. Tryptase was active following cleavage with a SUMOstar specific protease. The SUMOstar system will make significant impact in difficult-to-express proteins and especially to those proteins that require the native N-terminal residue for function.

  14. MaxSUMO: A New Expert Approach for Evaluating Mobility Management Projects

    Directory of Open Access Journals (Sweden)

    Veronique Van Acker

    2013-06-01

    Full Text Available There is need for better understanding of how mobility management interventions work and how they affect the individuals’ modal choice decisions, as well as need for robust evaluation techniques allowing any behavioural changes to be observed. Changing individual’s behaviour is not a one-step process and any evaluation methodology should account for this. A new standardized expert evaluation resource MaxSUMO takes this step-wise process into account. MaxSUMO is based on a new theoretical behavioural change model MaxSEM which measures individuals’ stage positions (their susceptibility to change behaviour and stage movement (progression towards actual behavioural change. This paper illustrates the use of MaxSUMO by the evaluation of the mobility campaign “I keep moving, even without my car” undertaken by the City of Ghent.

  15. Ubiquitin E3 ligase FIEL1 regulates fibrotic lung injury through SUMO-E3 ligase PIAS4.

    Science.gov (United States)

    Lear, Travis; McKelvey, Alison C; Rajbhandari, Shristi; Dunn, Sarah R; Coon, Tiffany A; Connelly, William; Zhao, Joe Y; Kass, Daniel J; Zhang, Yingze; Liu, Yuan; Chen, Bill B

    2016-05-30

    The E3 small ubiquitin-like modifier (SUMO) protein ligase protein inhibitor of activated STAT 4 (PIAS4) is a pivotal protein in regulating the TGFβ pathway. In this study, we discovered a new protein isoform encoded by KIAA0317, termed fibrosis-inducing E3 ligase 1 (FIEL1), which potently stimulates the TGFβ signaling pathway through the site-specific ubiquitination of PIAS4. FIEL1 targets PIAS4 using a double locking mechanism that is facilitated by the kinases PKCζ and GSK3β. Specifically, PKCζ phosphorylation of PIAS4 and GSK3β phosphorylation of FIEL1 are both essential for the degradation of PIAS4. FIEL1 protein is highly expressed in lung tissues from patients with idiopathic pulmonary fibrosis (IPF), whereas PIAS4 protein levels are significantly reduced. FIEL1 overexpression significantly increases fibrosis in a bleomycin murine model, whereas FIEL1 knockdown attenuates fibrotic conditions. Further, we developed a first-in-class small molecule inhibitor toward FIEL1 that is highly effective in ameliorating fibrosis in mice. This study provides a basis for IPF therapeutic intervention by modulating PIAS4 protein abundance.

  16. Characterisation of the SUMO-like domains of Schizosaccharomyces pombe Rad60.

    Directory of Open Access Journals (Sweden)

    Lara K Boyd

    Full Text Available The S. pombe Rad60 protein is required for the repair of DNA double strand breaks, recovery from replication arrest, and is essential for cell viability. It has two SUMO-like domains (SLDs at its C-terminus, an SXS motif and three sequences that have been proposed to be SUMO-binding motifs (SBMs. SMB1 is located in the middle of the protein, SBM2 is in SLD1 and SBM3 is at the C-terminus of SLD2. We have probed the functions of the two SUMO-like domains, SLD1 and SLD2, and the putative SBMs. SLD1 is essential for viability, while SLD2 is not. rad60-SLD2Δ cells are sensitive to DNA damaging agents and hydroxyurea. Neither ubiquitin nor SUMO can replace SLD1 or SLD2. Cells in which either SBM1 or SBM2 has been mutated are viable and are wild type for response to MMS and HU. In contrast mutation of SBM3 results in significant sensitivity to MMS and HU. These results indicate that the lethality resulting from deletion of SLD1 is not due to loss of SBM2, but that mutation of SBM3 produces a more severe phenotype than does deletion of SLD2. Using chemical denaturation studies, FPLC and dynamic light scattering we show this is likely due to the destabilisation of SLD2. Thus we propose that the region corresponding to the putative SBM3 forms part of the hydrophobic core of SLD2 and is not a SUMO-interacting motif. Over-expression of Hus5, which is the SUMO conjugating enzyme and known to interact with Rad60, does not rescue rad60-SLD2Δ, implying that as well as having a role in the sumoylation process as previously described, Rad60 has a Hus5-independent function.

  17. Enhanced in vitro refolding of fibroblast growth factor 15 with the assistance of SUMO fusion partner.

    Directory of Open Access Journals (Sweden)

    Bo Kong

    Full Text Available Fibroblast growth factor 15 (Fgf15 is the mouse orthologue of human FGF19. Fgf15 is highly expressed in the ileum and functions as an endocrine signal to regulate liver function, including bile acid synthesis, hepatocyte proliferation and insulin sensitivity. In order to fully understand the function of Fgf15, methods are needed to produce pure Fgf15 protein in the prokaryotic system. However, when expressed in Escherichia coli (E. coli, the recombinant Fgf15 protein was insoluble and found only in inclusion bodies. In the current study, we report a method to produce recombinant Fgf15 protein in E. coli through the use of small ubiquitin-related modifier (SUMO fusion tag. Even though the SUMO has been shown to strongly improve protein solubility and expression levels, our studies suggest that the SUMO does not improve Fgf15 protein solubility. Instead, proper refolding of Fgf15 protein was achieved when Fgf15 was expressed as a partner protein of the fusion tag SUMO, followed by in vitro dialysis refolding. After refolding, the N-terminal SUMO tag was cleaved from the recombinant Fgf15 fusion protein by ScUlp1 (Ubiquitin-Like Protein-Specific Protease 1 from S. cerevisiae. With or without the SUMO tag, the refolded Fgf15 protein was biologically active, as revealed by its ability to reduce hepatic Cyp7a1 mRNA levels in mice. In addition, recombinant Fgf15 protein suppressed Cyp7a1 mRNA levels in a dose-dependent manner. In summary, we have developed a successful method to express functional Fgf15 protein in prokaryotic cells.

  18. Anterior mediastinal abscess diagnosed in a young sumo wrestler after closed blunt chest trauma

    Institute of Scientific and Technical Information of China (English)

    Tatsuro Sassa; Ken-ichiro Kobayashi; Masayuki Ota; Takuya Washino; Mayu Hikone; Naoya Sakamoto; Sentaro Iwabuchi

    2015-01-01

    Most mediastinal abscesses result from infections after thoracotomy, esophageal perforation or penetrating chest trauma.This disease is rarely caused by closed blunt chest trauma.All previously reported such cases after closed blunt chest trauma presented with hematoma and sternal osteomyelitis resulting from sternal fracture.Here we report a 15-year-old sumo wrestler who presented with an anterior mediastinal abscess without any mediastinal fracture.The mediastinal abscess resulted from the hematogenous spread of Staphylococcus aureus to a hematoma that might have been caused by a closed blunt chest trauma incurred during sumo wrestling exercises.

  19. Redox regulation of SUMO enzymes is required for ATM activity and survival in oxidative stress.

    Science.gov (United States)

    Stankovic-Valentin, Nicolas; Drzewicka, Katarzyna; König, Cornelia; Schiebel, Elmar; Melchior, Frauke

    2016-06-15

    To sense and defend against oxidative stress, cells depend on signal transduction cascades involving redox-sensitive proteins. We previously identified SUMO (small ubiquitin-related modifier) enzymes as downstream effectors of reactive oxygen species (ROS). Hydrogen peroxide transiently inactivates SUMO E1 and E2 enzymes by inducing a disulfide bond between their catalytic cysteines. How important their oxidation is in light of many other redox-regulated proteins has however been unclear. To selectively disrupt this redox switch, we identified a catalytically fully active SUMO E2 enzyme variant (Ubc9 D100A) with strongly reduced propensity to maintain a disulfide with the E1 enzyme in vitro and in cells. Replacement of Ubc9 by this variant impairs cell survival both under acute and mild chronic oxidative stresses. Intriguingly, Ubc9 D100A cells fail to maintain activity of the ATM-Chk2 DNA damage response pathway that is induced by hydrogen peroxide. In line with this, these cells are also more sensitive to the ROS-producing chemotherapeutic drugs etoposide/Vp16 and Ara-C. These findings reveal that SUMO E1~E2 oxidation is an essential redox switch in oxidative stress.

  20. Modellering van begrazing in SUMO : verbetering van de vegetatiemodellering in de Natuurplanner

    NARCIS (Netherlands)

    Wamelink, G.W.W.; Wegman, R.M.A.; Slim, P.A.; Dirksen, J.; Mol-Dijkstra, J.P.; Dobben, van H.F.

    2001-01-01

    In samenwerking met het RIVM en WU wordt het vegetatievoorspellingsmodel SUMO ontwikkeld. Het model vormt een integraal onderdeel met SMART en wordt onder andere gebruikt in de natuurplanner van het RIVM. Om de modellering van de effecten van begrazing op de vegetatieontwikkeling en successie mogeli

  1. Functional expression and purification of recombinant Hepcidin25 production in Escherichia coli using SUMO fusion technology.

    Science.gov (United States)

    Sadr, Vahideh; Saffar, Behnaz; Emamzadeh, Rahman

    2017-04-30

    Hepcidin25 is a small cysteine-rich peptide hormone known as a new class of antimicrobial peptides. The purpose of the present study was to express, purify and investigate the antibacterial properties of recombinant human hepcidin25 protein production in Escherichia coli. Human hepcidin25 gene was optimized and fused to a small ubiquitin-related modifier (SUMO) gene for higher expression. Then SUMO-hepcidin25 was cloned into the pET-32a (+) vector and expressed in E. coli Origami. The fusion protein with a molecular weight of approximately 35kDa was analyzed on SDS-PAGE gel. The highest expression was observed after 6h induction and the fusion protein consisted approximately 47% of the total cellular protein. The purified SUMO-hepcidin25 purity was determined to be higher than 95%, with a final yield of 3.9mgl(-)(1) of media. The recombinant hepcidin25 showed antibacterial activity against both Gram negative (Klebsiella pneumonia) and Gram positive (Staphylococcus aureus and Bacillus cereus) bacteria with minimum inhibitory concentrations (MICs) of 150μgml(-1), 18.7μg/ml(-1) and 37.5μg/ml(-1), respectively. These results indicated that thioredoxin and SUMO dual fusion system is an efficient production system for synthesis functional human hepcidin25.

  2. Study of MDM2 and SUMO-1 expression in actinic cheilitis and lip cancer.

    Science.gov (United States)

    Oliveira Alves, Mônica Ghislaine; da Mota Delgado, Adriana; Balducci, Ivan; Carvalho, Yasmin Rodarte; Cavalcante, Ana Sueli Rodrigues; Almeida, Janete Dias

    2014-11-01

    Actinic cheilitis exhibits a potential of malignant transformation in 10-20 % of cases. The objective of this study was to compare the expression of MDM2 and SUMO-1 proteins between actinic cheilitis (AC) and squamous cell carcinoma (SCC) of the lip. The sample consisted of lower lip mucosa specimens obtained from cases with a clinical and histopathological diagnosis of AC (n = 26) and SCC (n = 25) and specimens of labial semi-mucosa (n = 15) without clinical alterations or inflammation. The tissue samples were stained with hematoxylin-eosin and anti-MDM2 and anti-SUMO-1 antibodies. Data were analyzed by the Kruskal-Wallis and Dunn's tests (5 %). The median expression of MDM2 (kW = 36.8565; df = 3-1 = 2; p = 0.0001) and SUMO-1 (kW = 32.7080; df = 3-1 = 2; p = 0.0001) was similar in cases of AC and SCC of the lip, but differed significantly from that observed for normal labial semi-mucosa. Despite the limitations of the present study, immunohistochemistry demonstrated the overexpression of important proteins (MDM2 and SUMO-1) related to regulatory mechanisms of apoptosis in AC and SCC of the lip, but further studies are needed.

  3. The Small Unmanned Meteorological Observer SUMO: A new tool for atmospheric boundary layer research

    Directory of Open Access Journals (Sweden)

    Joachim Reuder

    2009-05-01

    Full Text Available The Small Unmanned Meteorological Observer SUMO has been developed as a cost-efficient measurement system with the aim to close the existing observational gap of atmospheric measurement systems in between meteorological masts/towers and radiosondes. The system is highly flexible and has the capability for in-situ ABL measurements with unique spatial and temporal resolution. SUMO is based on a light-weighted styrofoam model airplane, equipped with an autopilot system for autonomous flight missions and in its recent version with meteorological sensors for temperature, humidity and pressure. With its wingspan of 80 cm, its length of 75 cm and a total lift-off weight of 580 g, SUMO is easy to transport and operate even in remote areas with limited infrastructure. During several field campaigns in 2007 and 2008 the system has been successfully tested and operated. Atmospheric profiles of temperature, humidity, wind speed and wind direction have been determined up to 3500 m above ground during the FLOHOF (FLOw over and around HOFsjökull field campaign in Central Iceland in July/August 2007. During a 3 week campaign on and around Spitsbergen in February/March 2008 the SUMO system also proved its functionality under polar conditions, reaching altitudes above 1500 m even at ground temperatures of -20° C and wind speeds up to 15 m s-1.

  4. The Small Unmanned Meteorological Observer SUMO. A new tool for atmospheric boundary layer research

    Energy Technology Data Exchange (ETDEWEB)

    Reuder, Joachim; Jonassen, Marius; Mayer, Stephanie [Bergen Univ. (Norway). Geophysical Inst.; Brisset, Pascal [Ecole Nationale de l' Aviation Civile (ENAC), Toulouse (France); Mueller, Martin [Martin Mueller Engineering, Hildesheim (Germany)

    2009-04-15

    The Small Unmanned Meteorological Observer SUMO has been developed as a cost-efficient measurement system with the aim to close the existing observational gap of atmospheric measurement systems in between meteorological masts/towers and radiosondes. The system is highly flexible and has the capability for in-situ ABL measurements with unique spatial and temporal resolution. SUMO is based on a light-weighted styrofoam model airplane, equipped with an autopilot system for autonomous flight missions and in its recent version with meteorological sensors for temperature, humidity and pressure. With its wingspan of 80 cm, its length of 75 cm and a total lift-off weight of 580 g, SUMO is easy to transport and operate even in remote areas with limited infrastructure. During several field campaigns in 2007 and 2008 the system has been successfully tested and operated. Atmospheric profiles of temperature, humidity, wind speed and wind direction have been determined up to 3500 m above ground during the FLOHOF (FLOw over and around HOFsjoekull) field campaign in Central Iceland in July/August 2007. During a 3 week campaign on and around Spitsbergen in February/March 2008 the SUMO system also proved its functionality under polar conditions, reaching altitudes above 1500 m even at ground temperatures of -20 C and wind speeds up to 15 m s{sup -1}. (orig.)

  5. El sumo bien kantiano: el objeto construído de la ley moral

    Directory of Open Access Journals (Sweden)

    Alonso Villarán

    2016-01-01

    Full Text Available El presente artículo tiene tres objetivos: a. compilar las críticas planteadas en contra del sumo bien kantiano (un mundo donde la virtud es premiada con felicidad; b. elaborar una revisión crítica de las respectivas defensas; y c. desarrollar una nueva defensa, con énfasis en el problema de derivación (según el cual el sumo bien no se deriva de la ley moral. Respecto al primero punto, el artículo muestra que son al menos cinco los problemas que amenazan la doctrina: heteronomía, derivación, imposibilidad, irrelevancia y dualismo. Respecto al segundo, el artículo revela que las múltiples defensas elaboradas en nombre del sumo bien son fuente, a su vez, de otros problemas, como los de incompletitud, ilusión, injusticia, deslealtad y escisión. Respecto al tercer y principal punto, y empezando por la derivación, el artículo propone ver al sumo bien como el objeto construido de la ley moral. Resuelto el problema de derivación, el artículo ofrece breves respuestas a los otros problemas en juego. El artículo también traduce por primera vez, del inglés al español, fragmentos de importantes publicaciones sobre el sumo bien.

  6. Sumoylation: a regulatory protein modification in health and disease.

    Science.gov (United States)

    Flotho, Annette; Melchior, Frauke

    2013-01-01

    Posttranslational modification with small ubiquitin-related modifier (SUMO) proteins is now established as one of the key regulatory protein modifications in eukaryotic cells. Hundreds of proteins involved in processes such as chromatin organization, transcription, DNA repair, macromolecular assembly, protein homeostasis, trafficking, and signal transduction are subject to reversible sumoylation. Hence, it is not surprising that disease links are beginning to emerge and that interference with sumoylation is being considered for intervention. Here, we summarize basic mechanisms and highlight recent developments in the physiology of sumoylation.

  7. SUMO-1 Promotes Ishikawa Cell Proliferation and Apoptosis in Endometrial Cancer by Increasing Sumoylation of Histone H4.

    Science.gov (United States)

    Zheng, Jindan; Liu, Lili; Wang, Shanfeng; Huang, Xin

    2015-10-01

    To investigate the functional role of SUMO-1 on cell proliferation and apoptosis in endometrial cancer cells (Ishikawa line) and to explore the underlying regulatory mechanisms. Different concentrations of estradiol (E2) and small interfering RNA (siRNA) targeting the SUMO-1 (siCo) were treated in Ishikawa cells, and then quantitative polymerase chain reaction was used to examine the expression of progesterone receptor (PR) expression in Ishikawa cells. Western blots were applied to validate histone H4 sumoylation. CCK8 assay and flow cytometry were performed to investigate cell proliferation and apoptosis in Ishikawa cells. Estradiol up-regulated the expression of PR messenger RNA, most obviously at 100 nM. SUMO-1 siRNA decreased the PR expression. Estradiol up-regulated H4 sumoylation and caused the increase of Ishikawa cell proliferation, whereas SUMO-1 siRNA down-regulated H4 sumoylation, inhibited the cell proliferation, and induced apoptosis. Furthermore, SUMO-1 siRNA-transfected cells were arrested in the S and G2/M phases and E2 increased the S and G2/M phases of Ishikawa cells. SUMO-1 regulates the Ishikawa cell proliferation and apoptosis by the sumoylation of histone H4.

  8. Slx8 removes Pli1-dependent protein-SUMO conjugates including SUMOylated topoisomerase I to promote genome stability.

    Directory of Open Access Journals (Sweden)

    Roland Steinacher

    Full Text Available The SUMO-dependent ubiquitin ligase Slx8 plays key roles in promoting genome stability, including the processing of trapped Topoisomerase I (Top1 cleavage complexes and removal of toxic SUMO conjugates. We show that it is the latter function that constitutes Slx8's primary role in fission yeast. The SUMO conjugates in question are formed by the SUMO ligase Pli1, which is necessary for limiting spontaneous homologous recombination when Top1 is present. Surprisingly there is no requirement for Pli1 to limit recombination in the vicinity of a replication fork blocked at the programmed barrier RTS1. Notably, once committed to Pli1-mediated SUMOylation Slx8 becomes essential for genotoxin resistance, limiting both spontaneous and RTS1 induced recombination, and promoting normal chromosome segregation. We show that Slx8 removes Pli1-dependent Top1-SUMO conjugates and in doing so helps to constrain recombination at RTS1. Overall our data highlight how SUMOylation and SUMO-dependent ubiquitylation by the Pli1-Slx8 axis contribute in different ways to maintain genome stability.

  9. Slx8 Removes Pli1-Dependent Protein-SUMO Conjugates Including SUMOylated Topoisomerase I to Promote Genome Stability

    Science.gov (United States)

    Steinacher, Roland; Osman, Fekret; Lorenz, Alexander; Bryer, Claire; Whitby, Matthew C.

    2013-01-01

    The SUMO-dependent ubiquitin ligase Slx8 plays key roles in promoting genome stability, including the processing of trapped Topoisomerase I (Top1) cleavage complexes and removal of toxic SUMO conjugates. We show that it is the latter function that constitutes Slx8's primary role in fission yeast. The SUMO conjugates in question are formed by the SUMO ligase Pli1, which is necessary for limiting spontaneous homologous recombination when Top1 is present. Surprisingly there is no requirement for Pli1 to limit recombination in the vicinity of a replication fork blocked at the programmed barrier RTS1. Notably, once committed to Pli1-mediated SUMOylation Slx8 becomes essential for genotoxin resistance, limiting both spontaneous and RTS1 induced recombination, and promoting normal chromosome segregation. We show that Slx8 removes Pli1-dependent Top1-SUMO conjugates and in doing so helps to constrain recombination at RTS1. Overall our data highlight how SUMOylation and SUMO-dependent ubiquitylation by the Pli1-Slx8 axis contribute in different ways to maintain genome stability. PMID:23936535

  10. Covalent conjugation of the equine infectious anemia virus Gag with SUMO.

    Science.gov (United States)

    Wang, Jinzhong; Wen, Shuping; Zhao, Rui; Qi, Jing; Liu, Zhao; Li, Weiwei; An, Jing; Wood, Charles; Wang, Ying

    2017-05-06

    The conjugation of small ubiquitin-like modifier (SUMO) to the target protein, namely, SUMOylation, is involved in the regulation of many important biological events including host-pathogen interaction. Some viruses have evolved to exploit the host SUMOylation machinery to modify their own protein. Retroviral Gag protein plays critical roles in the viral life cycle. The HIV-1 p6 and the Moloney murine leukemia virus CA have been reported to be conjugated with SUMO. In this study, we report for the first time, to our knowledge, the covalent conjugation of equine infectious anemia virus (EIAV) Gag with SUMO. The C-terminal p9 domain of Gag is a main target for SUMOylation and SUMO is attached to multiple sites of p9, including K30 whose mutation abolished p9 SUMOylation completely. The SUMOylation of p9, but not the p9-K30 mutant, was also detected in equine fibroblastic cells ATCC(®) CCL-57™. Ubc9 and its C93 residue are indispensable for the SUMOylation of p9. Using confocal microscopy, it is found that EIAV Gag localizes primarily, if not exclusively, in the cytoplasm of the cell and the co-localization of EIAV Gag with Ubc9 was observed. Our findings that EIAV Gag is SUMOylated at p9-K30, together with previous findings on the defects of p9-K30 mutant in viral DNA translocation from cytoplasm to the nucleus, suggests that SUMOylation of Gag may be involved in such functions. Copyright © 2017 Elsevier Inc. All rights reserved.

  11. Scavenger receptors mediate the role of SUMO and Ftz-f1 in Drosophila steroidogenesis.

    Directory of Open Access Journals (Sweden)

    Ana Talamillo

    2013-04-01

    Full Text Available SUMOylation participates in ecdysteroid biosynthesis at the onset of metamorphosis in Drosophila melanogaster. Silencing the Drosophila SUMO homologue smt3 in the prothoracic gland leads to reduced lipid content, low ecdysone titers, and a block in the larval-pupal transition. Here we show that the SR-BI family of Scavenger Receptors mediates SUMO functions. Reduced levels of Snmp1 compromise lipid uptake in the prothoracic gland. In addition, overexpression of Snmp1 is able to recover lipid droplet levels in the smt3 knockdown prothoracic gland cells. Snmp1 expression depends on Ftz-f1 (an NR5A-type orphan nuclear receptor, the expression of which, in turn, depends on SUMO. Furthermore, we show by in vitro and in vivo experiments that Ftz-f1 is SUMOylated. RNAi-mediated knockdown of ftz-f1 phenocopies that of smt3 at the larval to pupal transition, thus Ftz-f1 is an interesting candidate to mediate some of the functions of SUMO at the onset of metamorphosis. Additionally, we demonstrate that the role of SUMOylation, Ftz-f1, and the Scavenger Receptors in lipid capture and mobilization is conserved in other steroidogenic tissues such as the follicle cells of the ovary. smt3 knockdown, as well as ftz-f1 or Scavenger knockdown, depleted the lipid content of the follicle cells, which could be rescued by Snmp1 overexpression. Therefore, our data provide new insights into the regulation of metamorphosis via lipid homeostasis, showing that Drosophila Smt3, Ftz-f1, and SR-BIs are part of a general mechanism for uptake of lipids such as cholesterol, required during development in steroidogenic tissues.

  12. Arabidopsis SUMO protease ASP1 positively regulates flowering time partially through regulating FLC stability 

    KAUST Repository

    Kong, Xiangxiong

    2016-12-07

    The initiation of flowering is tightly regulated by the endogenous and environment signals, which is crucial for the reproductive success of flowering plants. It is well known that autonomous and vernalization pathways repress transcription of FLOWERING LOCUS C (FLC), a focal floral repressor, but how its protein stability is regulated remains largely unknown. Here, we found that mutations in a novel Arabidopsis SUMO protease 1 (ASP1) resulted in a strong late-flowering phenotype under long-days, but to a lesser extent under short-days. ASP1 localizes in the nucleus and exhibited a SUMO protease activity in vitro and in vivo. The conserved Cys-577 in ASP1 is critical for its enzymatic activity, as well as its physiological function in the regulation of flowering time. Genetic and gene expression analyses demonstrated that ASP1 promotes transcription of positive regulators of flowering, such as FT, SOC1 and FD, and may function in both CO-dependent photoperiod pathway and FLC-dependent pathways. Although the transcription level of FLC was not affected in the loss-of-function asp1 mutant, the protein stability of FLC was increased in the asp1 mutant. Taken together, this study identified a novel bona fide SUMO protease, ASP1, which positively regulates transition to flowering at least partly by repressing FLC protein stability.

  13. Regulation of mitotic spindle asymmetry by SUMO and the spindle-assembly checkpoint in yeast.

    Science.gov (United States)

    Leisner, Christian; Kammerer, Daniel; Denoth, Annina; Britschi, Mirjam; Barral, Yves; Liakopoulos, Dimitris

    2008-08-26

    During mitosis, the kinetochore microtubules capture and segregate chromosomes, and the astral microtubules position the spindle within the cell. Although the spindle is symmetric, proper positioning of the spindle in asymmetrically dividing cells generally correlates with the formation of morphologically and structurally distinct asters [1]. In budding yeast, the spindle-orientation proteins Kar9 and dynein decorate only one aster of the metaphase spindle and direct it toward the bud [2, 3]. The mechanisms controlling the distribution of Kar9 and dynein remain unclear. Here, we show that SUMO regulates astral-microtubule function in at least two ways. First, Kar9 was sumoylated in vivo. Sumoylation and Cdk1-dependent phosphorylation of Kar9 independently promoted Kar9 asymmetry on the spindle. Second, proper regulation of kinetochore function by SUMO was also required for Kar9 asymmetry. Indeed, activation of the spindle-assembly checkpoint (SAC) due to SUMO and kinetochore defects promoted symmetric redistribution of Kar9 in a Mad2-dependent manner. The control of Kar9 distribution by the SAC was independent of Kar9 sumoylation and phosphorylation. Together, our data reveal that three independent mechanisms contribute to Kar9 asymmetry: Cdk1-dependent phosphorylation, sumoylation, and SAC signaling. Hence, the two seemingly independent spindle domains, kinetochores and astral microtubules, function in a tightly coordinated fashion.

  14. Tandem SUMO fusion vectors for improving soluble protein expression and purification.

    Science.gov (United States)

    Guerrero, Fernando; Ciragan, Annika; Iwaï, Hideo

    2015-12-01

    Availability of highly purified proteins in quantity is crucial for detailed biochemical and structural investigations. Fusion tags are versatile tools to facilitate efficient protein purification and to improve soluble overexpression of proteins. Various purification and fusion tags have been widely used for overexpression in Escherichia coli. However, these tags might interfere with biological functions and/or structural investigations of the protein of interest. Therefore, an additional purification step to remove fusion tags by proteolytic digestion might be required. Here, we describe a set of new vectors in which yeast SUMO (SMT3) was used as the highly specific recognition sequence of ubiquitin-like protease 1, together with other commonly used solubility enhancing proteins, such as glutathione S-transferase, maltose binding protein, thioredoxin and trigger factor for optimizing soluble expression of protein of interest. This tandem SUMO (T-SUMO) fusion system was tested for soluble expression of the C-terminal domain of TonB from different organisms and for the antiviral protein scytovirin.

  15. Posttranslational Modification Assays on Functional Protein Microarrays.

    Science.gov (United States)

    Neiswinger, Johnathan; Uzoma, Ijeoma; Cox, Eric; Rho, HeeSool; Jeong, Jun Seop; Zhu, Heng

    2016-10-03

    Protein microarray technology provides a straightforward yet powerful strategy for identifying substrates of posttranslational modifications (PTMs) and studying the specificity of the enzymes that catalyze these reactions. Protein microarray assays can be designed for individual enzymes or a mixture to establish connections between enzymes and substrates. Assays for four well-known PTMs-phosphorylation, acetylation, ubiquitylation, and SUMOylation-have been developed and are described here for use on functional protein microarrays. Phosphorylation and acetylation require a single enzyme and are easily adapted for use on an array. The ubiquitylation and SUMOylation cascades are very similar, and the combination of the E1, E2, and E3 enzymes plus ubiquitin or SUMO protein and ATP is sufficient for in vitro modification of many substrates.

  16. SUMO-fusion, purification, and characterization of a (+)-zizaene synthase from Chrysopogon zizanioides

    Energy Technology Data Exchange (ETDEWEB)

    Hartwig, S.; Frister, T.; Alemdar, S.; Li, Z.; Scheper, T.; Beutel, S., E-mail: beutel@iftc.uni-hannover.de

    2015-03-20

    An uncharacterized plant cDNA coding for a polypeptide presumably having sesquiterpene synthase activity, was expressed in soluble and active form. Two expression strategies were evaluated in Escherichia coli. The enzyme was fused to a highly soluble SUMO domain, in addition to being produced in an unfused form by a cold-shock expression system. Yields up to ∼325 mg/L{sup −1} were achieved in batch cultivations. The 6x-His-tagged enzyme was purified employing an Ni{sup 2+}-IMAC-based procedure. Identity of the protein was established by Western Blot analysis as well as peptide mass fingerprinting. A molecular mass of 64 kDa and an isoelectric point of pI 4.95 were determined by 2D gel electrophoresis. Cleavage of the fusion domain was possible by digestion with specific SUMO protease. The synthase was active in Mg{sup 2+} containing buffer and catalyzed the production of (+)-zizaene (syn. khusimene), a precursor of khusimol, from farnesyl diphosphate. Product identity was confirmed by GC–MS and comparison of retention indices. Enzyme kinetics were determined by measuring initial reaction rates for the product, using varying substrate concentrations. By assuming a Michaelis–Menten model, kinetic parameters of K{sub M} = 1.111 μM (±0.113), v{sub max} = 0.3245 μM min{sup −1} (±0.0035), k{sub cat} = 2.95 min{sup −1}, as well as a catalytic efficiency k{sub cat}/K{sub M} = 4.43 × 10{sup 4} M{sup −1} s{sup −1} were calculated. Fusion to a SUMO moiety can substantially increase soluble expression levels of certain hard to express terpene synthases in E. coli. The kinetic data determined for the recombinant synthase are comparable to other described plant sesquiterpene synthases and in the typical range of enzymes belonging to the secondary metabolism. This leaves potential for optimizing catalytic parameters through methods like directed evolution. - Highlights: • Uncharacterized (+)-zizaene synthase from C. zizanoides was cloned

  17. Stability of small ubiquitin-like modifier (SUMO) proteases OVERLY TOLERANT TO SALT1 and -2 modulates salicylic acid signalling and SUMO1/2 conjugation in Arabidopsis thaliana.

    Science.gov (United States)

    Bailey, Mark; Srivastava, Anjil; Conti, Lucio; Nelis, Stuart; Zhang, Cunjin; Florance, Hannah; Love, Andrew; Milner, Joel; Napier, Richard; Grant, Murray; Sadanandom, Ari

    2016-01-01

    Small ubiquitin-like modifier proteases 1 and 2 (SUMO1/2) have been linked to the regulation of salicylic acid (SA)-mediated defence signalling in Arabidopsis thaliana. In order to define the role of the SUMO proteases OVERLY TOLERANT TO SALT1 and -2 (OTS1/2) in defence and to provide insight into SUMO1/2-mediated regulation of SA signalling, we examined the status of SA-mediated defences in ots1/2 mutants. The ots1 ots2 double mutant displayed enhanced resistance to virulent Pseudomonas syringae and higher levels of SA compared with wild-type (WT) plants. Furthermore, ots1 ots2 mutants exhibited upregulated expression of the SA biosynthesis gene ICS1 in addition to enhanced SA-responsive ICS1 expression beyond that of WT. SA stimulated OTS1/2 degradation and promoted accumulation of SUMO1/2 conjugates. These results indicate that OTS1 and -2 act in a feedback loop in SA signalling and that de novo OTS1/2 synthesis works antagonistically to SA-promoted degradation, adjusting the abundance of OTS1/2 to moderate SA signalling. Accumulation of SUMO1/2 conjugates coincides with SA-promoted OTS degradation and may play a positive role in SA-mediated signalling in addition to its repressive roles reported elsewhere.

  18. Allosteric activation of SENP1 by SUMO1 β-grasp domain involves a dock-and-coalesce mechanism.

    Science.gov (United States)

    Guo, Jingjing; Zhou, Huan-Xiang

    2016-08-31

    Small ubiquitin-related modifiers (SUMOs) are conjugated to proteins to regulate a variety of cellular processes. SENPs are cysteine proteases with a catalytic center located within a channel between two subdomains that catalyzes SUMO C-terminal cleavage for processing of SUMO precursors and de-SUMOylation of target proteins. The β-grasp domain of SUMOs binds to an exosite cleft, and allosterically activates SENPs via an unknown mechanism. Our molecular dynamics simulations showed that binding of the β-grasp domain induces significant conformational and dynamic changes in SENP1, including widening of the exosite cleft and quenching of nanosecond dynamics in all but a distal region. A dock-and-coalesce mechanism emerges for SENP-catalyzed SUMO cleavage: the wedging of the β-grasp domain enables the docking of the proximal portion of the C-terminus and the strengthened cross-channel motional coupling initiates inter-subdomain correlated motions to allow for the distal portion to coalesce around the catalytic center.

  19. Characterization of the SUMO-binding activity of the myeloproliferative and mental retardation (MYM-type zinc fingers in ZNF261 and ZNF198.

    Directory of Open Access Journals (Sweden)

    Catherine M Guzzo

    Full Text Available SUMO-binding proteins interact with SUMO modified proteins to mediate a wide range of functional consequences. Here, we report the identification of a new SUMO-binding protein, ZNF261. Four human proteins including ZNF261, ZNF198, ZNF262, and ZNF258 contain a stretch of tandem zinc fingers called myeloproliferative and mental retardation (MYM-type zinc fingers. We demonstrated that MYM-type zinc fingers from ZNF261 and ZNF198 are necessary and sufficient for SUMO-binding and that individual MYM-type zinc fingers function as SUMO-interacting motifs (SIMs. Our binding studies revealed that the MYM-type zinc fingers from ZNF261 and ZNF198 interact with the same surface on SUMO-2 recognized by the archetypal consensus SIM. We also present evidence that MYM-type zinc fingers in ZNF261 contain zinc, but that zinc is not required for SUMO-binding. Immunofluorescence microscopy studies using truncated fragments of ZNF198 revealed that MYM-type zinc fingers of ZNF198 are necessary for localization to PML-nuclear bodies (PML-NBs. In summary, our studies have identified and characterized the SUMO-binding activity of the MYM-type zinc fingers in ZNF261 and ZNF198.

  20. DNA damage-inducible SUMOylation of HERC2 promotes RNF8 binding via a novel SUMO-binding Zinc finger

    DEFF Research Database (Denmark)

    Danielsen, Jannie Michaela Rendtlew; Povlsen, Lou Klitgaard; Villumsen, Bine Hare

    2012-01-01

    Nonproteolytic ubiquitylation of chromatin surrounding deoxyribonucleic acid (DNA) double-strand breaks (DSBs) by the RNF8/RNF168/HERC2 ubiquitin ligases facilitates restoration of genome integrity by licensing chromatin to concentrate genome caretaker proteins near the lesions. In parallel......, SUMOylation of so-far elusive upstream DSB regulators is also required for execution of this ubiquitin-dependent chromatin response. We show that HERC2 and RNF168 are novel DNA damage-dependent SUMOylation targets in human cells. In response to DSBs, both HERC2 and RNF168 were specifically modified with SUMO1...... at DSB sites in a manner dependent on the SUMO E3 ligase PIAS4. SUMOylation of HERC2 was required for its DSB-induced association with RNF8 and for stabilizing the RNF8-Ubc13 complex. We also demonstrate that the ZZ Zinc finger in HERC2 defined a novel SUMO-specific binding module, which together...

  1. SUMO-fusion, purification, and characterization of a (+)-zizaene synthase from Chrysopogon zizanioides.

    Science.gov (United States)

    Hartwig, S; Frister, T; Alemdar, S; Li, Z; Scheper, T; Beutel, S

    2015-03-20

    An uncharacterized plant cDNA coding for a polypeptide presumably having sesquiterpene synthase activity, was expressed in soluble and active form. Two expression strategies were evaluated in Escherichia coli. The enzyme was fused to a highly soluble SUMO domain, in addition to being produced in an unfused form by a cold-shock expression system. Yields up to ∼325 mg/L(-1) were achieved in batch cultivations. The 6x-His-tagged enzyme was purified employing an Ni(2+)-IMAC-based procedure. Identity of the protein was established by Western Blot analysis as well as peptide mass fingerprinting. A molecular mass of 64 kDa and an isoelectric point of pI 4.95 were determined by 2D gel electrophoresis. Cleavage of the fusion domain was possible by digestion with specific SUMO protease. The synthase was active in Mg(2+) containing buffer and catalyzed the production of (+)-zizaene (syn. khusimene), a precursor of khusimol, from farnesyl diphosphate. Product identity was confirmed by GC-MS and comparison of retention indices. Enzyme kinetics were determined by measuring initial reaction rates for the product, using varying substrate concentrations. By assuming a Michaelis-Menten model, kinetic parameters of KM = 1.111 μM (±0.113), vmax = 0.3245 μM min(-1) (±0.0035), kcat = 2.95 min(-1), as well as a catalytic efficiency kcat/KM = 4.43 × 10(4) M(-1)s(-1) were calculated. Fusion to a SUMO moiety can substantially increase soluble expression levels of certain hard to express terpene synthases in E. coli. The kinetic data determined for the recombinant synthase are comparable to other described plant sesquiterpene synthases and in the typical range of enzymes belonging to the secondary metabolism. This leaves potential for optimizing catalytic parameters through methods like directed evolution. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Proof of concept for turbulence measurements with the RPAS SUMO during the BLLAST campaign

    Science.gov (United States)

    Båserud, Line; Reuder, Joachim; Jonassen, Marius O.; Kral, Stephan T.; Paskyabi, Mostafa B.; Lothon, Marie

    2016-10-01

    The micro-RPAS (remotely piloted aircraft system) SUMO (Small Unmanned Meteorological Observer) equipped with a five-hole-probe (5HP) system for turbulent flow measurements was operated in 49 flight missions during the BLLAST (Boundary-Layer Late Afternoon and Sunset Turbulence) field campaign in 2011. Based on data sets from these flights, we investigate the potential and limitations of airborne velocity variance and TKE (turbulent kinetic energy) estimations by an RPAS with a take-off weight below 1 kg. The integration of the turbulence probe in the SUMO system was still in an early prototype stage during this campaign, and therefore extensive post-processing of the data was required. In order to be able to calculate the three-dimensional wind vector, flow probe measurements were first synchronized with the autopilot's attitude and velocity data. Clearly visible oscillations were detected in the resulting vertical velocity, w, even after correcting for the aircraft motion. The oscillations in w were identified as the result of an internal time shift between the inertial measurement unit (IMU) and the GPS sensors, leading to insufficient motion correction, especially for the vertical wind component, causing large values of σw. Shifting the IMU 1-1.5 s forward in time with respect to the GPS yields a minimum for σw and maximum covariance between the IMU pitch angle and the GPS climb angle. The SUMO data show a good agreement to sonic anemometer data from a 60 m tower for σu, but show slightly higher values for σv and σw. Vertical TKE profiles, obtained from consecutive flight legs at different altitudes, show reasonable results, both with respect to the overall TKE level and the temporal variation. A thorough discussion of the methods used and the identified uncertainties and limitations of the system for turbulence measurements is included and should help the developers and users of other systems with similar problems.

  3. PERFORMANCE ANALYSIS OF COMPOSITE LEAF SPRING IN A DEFENCE SUMO VEHICLE

    OpenAIRE

    M. Venkatesan; V. C. SATHISH GANDHI; E. JANARTHAN

    2015-01-01

    The composite material has taking place a major role in an automobiles industries. The leaf spring, which is considered for this study is a specially designed leaf spring used in SUMO design by the ordinance factory. The leaf spring which is an automotive component used to absorb vibrations induced during the motion of vehicle. It also acts as a structure to support vertical loading due to the weight of the vehicle and payload. In this study the Finite element method is used for analysing ...

  4. Kaposi's sarcoma-associated herpesvirus K-Rta exhibits SUMO-targeting ubiquitin ligase (STUbL like activity and is essential for viral reactivation.

    Directory of Open Access Journals (Sweden)

    Yoshihiro Izumiya

    Full Text Available The small ubiquitin-like modifier (SUMO is a protein that regulates a wide variety of cellular processes by covalent attachment of SUMO moieties to a diverse array of target proteins. Sumoylation also plays an important role in the replication of many viruses. Previously, we showed that Kaposi's sarcoma-associated herpesvirus (KSHV encodes a SUMO-ligase, K-bZIP, which catalyzes sumoylation of host and viral proteins. We report here that this virus also encodes a gene that functions as a SUMO-targeting ubiquitin-ligase (STUbL which preferentially targets sumoylated proteins for degradation. K-Rta, the major transcriptional factor which turns on the entire lytic cycle, was recently found to have ubiquitin ligase activity toward a selected set of substrates. We show in this study that K-Rta contains multiple SIMs (SUMO interacting motif and binds SUMOs with higher affinity toward SUMO-multimers. Like RNF4, the prototypic cellular STUbL, K-Rta degrades SUMO-2/3 and SUMO-2/3 modified proteins, including promyelocytic leukemia (PML and K-bZIP. PML-NBs (nuclear bodies or ND-10 are storage warehouses for sumoylated proteins, which negatively regulate herpesvirus infection, as part of the intrinsic immune response. Herpesviruses have evolved different ways to degrade or disperse PML bodies, and KSHV utilizes K-Rta to inhibit PML-NBs formation. This process depends on K-Rta's ability to bind SUMO, as a K-Rta SIM mutant does not effectively degrade PML. Mutations in the K-Rta Ring finger-like domain or SIM significantly inhibited K-Rta transactivation activity in reporter assays and in the course of viral reactivation. Finally, KSHV with a mutation in the Ring finger-like domain or SIM of K-Rta replicates poorly in culture, indicating that reducing SUMO-conjugates in host cells is important for viral replication. To our knowledge, this is the first virus which encodes both a SUMO ligase and a SUMO-targeting ubiquitin ligase that together may generate

  5. Structural Analysis of a Complex between Small Ubiquitin-like Modifier 1 (SUMO1) and the ZZ Domain of CREB-binding Protein (CBP/p300) Reveals a New Interaction Surface on SUMO

    DEFF Research Database (Denmark)

    Diehl, Carl; Akke, Mikael; Bekker-Jensen, Simon;

    2016-01-01

    )N Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersions, which define the off rates for the ZZ domain and SIM motif and show that the dynamic binding process has different characteristics for the two cases. Furthermore, in the absence of bound ligands SUMO1 transiently samples a high energy...

  6. Role of SUMO-specific protease 2 in reprogramming cellular glucose metabolism.

    Directory of Open Access Journals (Sweden)

    Shuang Tang

    Full Text Available Most cancer cells exhibit a shift in glucose metabolic strategy, displaying increased glycolysis even with adequate oxygen supply. SUMO-specific proteases (SENPs de-SUMOylate substrates including HIF1α and p53,two key regulators in cancer glucose metabolism, to regulate their activity, stability and subcellular localization. However, the role of SENPs in tumor glucose metabolism remains unclear. Here we report that SUMO-specific protease 2 (SENP2 negatively regulates aerobic glycolysis in MCF7 and MEF cells. Over-expression of SENP2 reduces the glucose uptake and lactate production, increasing the cellular ATP levels in MCF7 cells, while SENP2 knockout MEF cells show increased glucose uptake and lactate production along with the decreased ATP levels. Consistently, the MCF7 cells over-expressing SENP2 exhibit decreased expression levels of key glycolytic enzymes and an increased rate of glucose oxidation compared with control MCF7 cells, indicating inhibited glycolysis but enhanced oxidative mitochondrial respiration. Moreover, SENP2 over-expressing MCF7 cells demonstrated a reduced amount of phosphorylated AKT, whereas SENP2 knockout MEFs exhibit increased levels of phosphorylated AKT. Furthermore, inhibiting AKT phosphorylation by LY294002 rescued the phenotype induced by SENP2 deficiency in MEFs. In conclusion, SENP2 represses glycolysis and shifts glucose metabolic strategy, in part through inhibition of AKT phosphorylation. Our study reveals a novel function of SENP2 in regulating glucose metabolism.

  7. Antagonistic roles of ubiquitin ligase HEI10 and SUMO ligase RNF212 regulate meiotic recombination.

    Science.gov (United States)

    Qiao, Huanyu; Prasada Rao, H B D; Yang, Ye; Fong, Jared H; Cloutier, Jeffrey M; Deacon, Dekker C; Nagel, Kathryn E; Swartz, Rebecca K; Strong, Edward; Holloway, J Kim; Cohen, Paula E; Schimenti, John; Ward, Jeremy; Hunter, Neil

    2014-02-01

    Crossover recombination facilitates the accurate segregation of homologous chromosomes during meiosis. In mammals, poorly characterized regulatory processes ensure that every pair of chromosomes obtains at least one crossover, even though most recombination sites yield non-crossovers. Designation of crossovers involves selective localization of the SUMO ligase RNF212 to a minority of recombination sites, where it stabilizes pertinent factors such as MutSγ (ref. 4). Here we show that the ubiquitin ligase HEI10 (also called CCNB1IP1) is essential for this crossover/non-crossover differentiation process. In HEI10-deficient mice, RNF212 localizes to most recombination sites, and dissociation of both RNF212 and MutSγ from chromosomes is blocked. Consequently, recombination is impeded, and crossing over fails. In wild-type mice, HEI10 accumulates at designated crossover sites, suggesting that it also has a late role in implementing crossing over. As with RNF212, dosage sensitivity for HEI10 indicates that it is a limiting factor for crossing over. We suggest that SUMO and ubiquitin have antagonistic roles during meiotic recombination that are balanced to effect differential stabilization of recombination factors at crossover and non-crossover sites.

  8. PERFORMANCE ANALYSIS OF COMPOSITE LEAF SPRING IN A DEFENCE SUMO VEHICLE

    Directory of Open Access Journals (Sweden)

    M. VENKATESAN

    2015-05-01

    Full Text Available The composite material has taking place a major role in an automobiles industries. The leaf spring, which is considered for this study is a specially designed leaf spring used in SUMO design by the ordinance factory. The leaf spring which is an automotive component used to absorb vibrations induced during the motion of vehicle. It also acts as a structure to support vertical loading due to the weight of the vehicle and payload. In this study the Finite element method is used for analysing the composite spring for different parameters such us stress, deformation and mode frequencies for three different ratios of epoxy and E fiberglass materials. The composite specimen has been made in the three different ratios of material combination by hand layout moulding technique. The three different samples are 40% epoxy and 60% Efiberglass, 60% epoxy and 40% E-fiberglass, 70% epoxy and 30% E-fiberglass. The experiments were carried out for different test like tensile test, flexural test and hardness test. The experimental results are well within the simulation results and identified that the 40% epoxy and 60% E-fiberg as composite leaf spring is suitable for designing a spring in SUMO vehicle.

  9. Centromere binding and a conserved role in chromosome stability for SUMO-dependent ubiquitin ligases.

    Directory of Open Access Journals (Sweden)

    Loes A L van de Pasch

    Full Text Available The Saccharomyces cerevisiae Slx5/8 complex is the founding member of a recently defined class of SUMO-targeted ubiquitin ligases (STUbLs. Slx5/8 has been implicated in genome stability and transcription, but the precise contribution is unclear. To characterise Slx5/8 function, we determined genome-wide changes in gene expression upon loss of either subunit. The majority of mRNA changes are part of a general stress response, also exhibited by mutants of other genome integrity pathways and therefore indicative of an indirect effect on transcription. Genome-wide binding analysis reveals a uniquely centromeric location for Slx5. Detailed phenotype analyses of slx5Δ and slx8Δ mutants show severe mitotic defects that include aneuploidy, spindle mispositioning, fish hooks and aberrant spindle kinetics. This is associated with accumulation of the PP2A regulatory subunit Rts1 at centromeres prior to entry into anaphase. Knockdown of the human STUbL orthologue RNF4 also results in chromosome segregation errors due to chromosome bridges. The study shows that STUbLs have a conserved role in maintenance of chromosome stability and links SUMO-dependent ubiquitination to a centromere-specific function during mitosis.

  10. PIC-1/SUMO-1-Modified PML-Retinoic Acid Receptor α Mediates Arsenic Trioxide-Induced Apoptosis in Acute Promyelocytic Leukemia

    Science.gov (United States)

    Sternsdorf, Thomas; Puccetti, Elena; Jensen, Kirsten; Hoelzer, Dieter; Will, Hans; Ottmann, Oliver Gerhard; Ruthardt, Martin

    1999-01-01

    Fusion proteins involving the retinoic acid receptor α (RARα) and PML or PLZF nuclear protein are the genetic markers of acute promyelocytic leukemia (APL). APLs with PML-RARα or PLZF-RARα fusion protein differ only in their response to retinoic acid (RA) treatment: the t(15;17) (PML-RARα-positive) APL blasts are sensitive to RA in vitro, and patients enter disease remission after RA treatment, while those with t(11;17) (PLZF-RARα-positive) APLs do not. Recently it has been shown that complete remission can be achieved upon treatment with arsenic trioxide (As2O3) in PML-RARα-positive APL, even when the patient has relapsed and the disease is RA resistant. This appears to be due to apoptosis induced by As2O3 in the APL blasts by poorly defined mechanisms. Here we report that (i) As2O3 induces apoptosis only in cells expressing the PML-RARα, not the PLZF-RARα, fusion protein; (ii) PML-RARα is partially modified by covalent linkage with a PIC-1/SUMO-1-like protein prior to As2O3 treatment, whereas PLZF-RARα is not; (iii) As2O3 treatment induces a change in the modification pattern of PML-RARα toward highly modified forms; (iv) redistribution of PML nuclear bodies (PML-NBs) upon As2O3 treatment is accompanied by recruitment of PIC-1/SUMO-1 into PML-NBs, probably due to hypermodification of both PML and PML-RARα; (v) As2O3-induced apoptosis is independent of the DNA binding activity located in the RARα portion of the PML-RARα fusion protein; and (vi) the apoptotic process is bcl-2 and caspase 3 independent and is blocked only partially by a global caspase inhibitor. Taken together, these data provide novel insights into the mechanisms involved in As2O3-induced apoptosis in APL and predict that treatment of t(11;17) (PLZF-RARα-positive) APLs with As2O3 will not be successful. PMID:10373566

  11. The SUMOylation of Kif18A and its regulatory mechanism%Kif18A 的 SUMO 化修饰及其调控机制

    Institute of Scientific and Technical Information of China (English)

    李艳; 夏南松; 左勇

    2015-01-01

    Objective To investigate the SUMOylation of Kif18A and its regulatory mechanism.Methods Plas-mids Flag -Kif18A and HA -SUMO1 or His -SUMO1 were co -transfected into HEK293T cells.Flag -Kif18A proteins were pulled down by Flag M2 beads and its SUMOylation was detected by immunoblotting with anti -HA.His -SUMO1 conjugated proteins were also precipitated by TALON Resin and the level of SUMOylated Kif18A was detected by immno-blotting with Flag antibody.The potential sites of Kif18A SUMOlation were predicted by SUMOsp 2.0 software and its coding sequences were mutated from lysine to arginine on the Flag -Kif18A plasmid.Kif18A wide and mutant types were separately expressed in HEK293T cells.The change in the quantity of SUMOylated Kif18A was adopted to determine the site of SUMOylation in Flag -Kif18A.Furthermore, plasmids Flag -Kif18A, HA -SUMO1 and SENP1 WT or mutant were co -transfected into HEK293T cells.Then the amount of SUMOylated Kif18A was detected by immunoprecipitation to determine whether Kif18A can be de -SUMOylated by SENP1.Finally, HeLa cells were synchronized in G2 /M phase by Nocodazole treatment.The SUMOylation of Kif18A in HeLa cells in G2 /M phase were detected by immunoprecipitati-on and immunobloting.Results Kif18A could be SUMOylated by SUMO1 or SUMO2.K47 and K148 were two impor-tant sites for Kif18A SUMOylation.Besides, Kif18A could be de -SUMOylated by SENP1.The SUMOylation of Kif18A was decreased in HeLa cells arrested in G2 /M phase.Conclusion Kif18A can be SUMOylated and its SUMOylation is regulated by mitosis progression.%目的:探讨 Kif18A 的 SUMO 化修饰及其调控机制。方法在 HEK293T 细胞中共转染 Flag -Kif18A、HA -SUMO1或 His -SUMO1质粒,免疫沉淀富集 Flag -Kif18A,免疫印迹检测 HA -SUMO;用 TALON 树脂富集His -SUMO1蛋白,免疫印迹检测 Flag -Kif18A 考察 Kif18A 的 SUMO 化水平。利用 SUMOsp 2.0软件预测Kif18A 可能发生 SUMO 化的潜在位点,将 Flag -Kif18A

  12. The ubiquitin- and SUMO-dependent signaling response to DNA double-strand breaks

    DEFF Research Database (Denmark)

    Bekker-Jensen, Simon; Mailand, Niels

    2011-01-01

    DNA double-strand breaks (DSBs) represent the most destructive type of chromosomal lesion and trigger rapid chromatin restructuring accompanied by accumulation of proteins in the vicinity of the DSB. Non-proteolytic ubiquitylation of chromatin surrounding DSBs, mediated by the RNF8/RNF168 ubiquitin...... ligase cascade, has emerged as a key mechanism for restoration of genome integrity by licensing the DSB-modified chromatin to concentrate genome caretaker proteins such as 53BP1 and BRCA1 near the lesions. In parallel, SUMOylation of upstream DSB regulators is also required for execution...... of this ubiquitin-dependent chromatin response, but its molecular basis is currently unclear. Here, we discuss recent insights into how ubiquitin- and SUMO-dependent signaling processes cooperate to orchestrate protein interactions with sites of DNA damage to facilitate DSB repair....

  13. COMPARISON OF NORMALIZED MAXIMUM AEROBIC CAPACITY AND BODY COMPOSITION OF SUMO WRESTLERS TO ATHLETES IN COMBAT AND OTHER SPORTS

    Directory of Open Access Journals (Sweden)

    Matthew D. Beekley

    2006-07-01

    Full Text Available Sumo wrestling is unique in combat sport, and in all of sport. We examined the maximum aerobic capacity and body composition of sumo wrestlers and compared them to untrained controls. We also compared "aerobic muscle quality", meaning VO2max normalized to predicted skeletal muscle mass (SMM (VO2max /SMM, between sumo wrestlers and controls and among previously published data for male athletes from combat, aerobic, and power sports. Sumo wrestlers, compared to untrained controls, had greater (p < 0.05 body mass (mean ± SD; 117.0 ± 4.9 vs. 56.1 ± 9.8 kg, percent fat (24.0 ± 1.4 vs. 13.3 ± 4.5, fat-free mass (88.9 ± 4.2 vs. 48.4 �� 6.8 kg, predicted SMM (48.2 ± 2.9 vs. 20.6 ± 4.7 kg and absolute VO2max (3.6 ± 1.3 vs. 2.5 ± 0.7 L·min-1. Mean VO2max /SMM (ml·kg SMM-1·min-1 was significantly different (p < 0.05 among aerobic athletes (164.8 ± 18.3, combat athletes (which was not different from untrained controls; 131.4 ± 9.3 and 128.6 ± 13.6, respectively, power athletes (96.5 ± 5.3, and sumo wrestlers (71.4 ± 5.3. There was a strong negative correlation (r = - 0.75 between percent body fat and VO2max /SMM (p < 0.05. We conclude that sumo wrestlers have some of the largest percent body fat and fat-free mass and the lowest "aerobic muscle quality" (VO2max /SMM, both in combat sport and compared to aerobic and power sport athletes. Additionally, it appears from analysis of the relationship between SMM and absolute VO2max for all sports that there is a "ceiling" at which increases in SMM do not result in additional increases in absolute VO2max

  14. High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Huang Yadong

    2010-02-01

    Full Text Available Abstract Background Fibroblast growth factor 21 (FGF21 is a promising drug candidate to combat metabolic diseases. However, high-level expression and purification of recombinant FGF21 (rFGF21 in Escherichia coli (E. coli is difficult because rFGF21 forms inclusion bodies in the bacteria making it difficult to purify and obtain high concentrations of bioactive rFGF21. To overcome this problem, we fused the FGF21 with SUMO (Small ubiquitin-related modifier by polymerase chain reaction (PCR, and expressed the fused gene in E. coli BL21(DE3. Results By inducing with IPTG, SUMO-FGF21 was expressed at a high level. Its concentration reached 30% of total protein, and exceeded 95% of all soluble proteins. The fused protein was purified by DEAE sepharose FF and Ni-NTA affinity chromatography. Once cleaved by the SUMO protease, the purity of rFGF21 by high performance liquid chromatography (HPLC was shown to be higher than 96% with low endotoxin level (in vivo animal experiments showed that rFGF21 produced by using this method, could decrease the concentration of plasma glucose in diabetic rats by streptozotocin (STZ injection. Conclusions This study demonstrated that SUMO, when fused with FGF21, was able to promote its soluble expression of the latter in E. coli, making it more convenient to purify rFGF21 than previously. This may be a better method to produce rFGF21 for pharmaceutical research and development.

  15. Exploring the potential of the RPA system SUMO for multipurpose boundary-layer missions during the BLLAST campaign

    Science.gov (United States)

    Reuder, Joachim; Båserud, Line; Jonassen, Marius O.; Kral, Stephan T.; Müller, Martin

    2016-06-01

    In June and July 2011 the RPAS (Remotely Piloted Aircraft System) SUMO (Small Unmanned Meteorological Observer) performed a total number of 299 scientific flights during the BLLAST (Boundary Layer Late Afternoon and Sunset Turbulence) campaign in southern France. Three different types of missions were performed: vertical profiling of the mean meteorological parameters (temperature, humidity and wind), horizontal surveys of the surface temperature and horizontal transects for the estimation of turbulence. The manuscript provides an introduction to the corresponding SUMO operations, including regulatory issues and the coordination of manned and unmanned airborne operations for boundary-layer research that have been pioneered during the BLLAST campaign. The main purpose of the SUMO flight strategy was atmospheric profiling at high temporal resolution. A total of 168 profile flights were performed during the campaign with typically more than 10 flights per Intensive Observational Period (IOP) day. The collected data allow for a detailed study of boundary-layer structure and dynamics and will be used for further analysis, e.g. the determination of profiles of sensible and latent heat fluxes. First, tests of a corresponding method have shown very promising results and have provided surface-flux values in close agreement with those from ground-based eddy-covariance measurements. In addition, 74 horizontal surveys of the IR emission of the surface were performed at altitudes of around 65 m. Each of those surveys covers a typical area of around 1 km2 and allows for an estimation of the surface-temperature variability, important information for the assessment of the heterogeneity of the surface forcing as a function of soil and vegetation properties. The comparison with other surface-temperature measurements shows that the raw data of the airborne and ground observations can differ considerably, but that even a very simple multiple regression method can reduce those

  16. SUMO: solar ultraviolet monitor and ozone nanosatellite for spectral irradiance, ozone and Earth radiative budget simultaneous evaluation

    Science.gov (United States)

    Damé, Luc

    SUMO is an innovative proof-of-concept nano-satellite which aims to measure on the same platform the different components of the Earth radiation budget, the solar energy input and the energy reemitted at the top of the Earth atmosphere, with a particular focus on the UV part of the spectrum and on the ozone layer, which are the most sensitive to the solar variability. The far UV (FUV) is the only wavelength band with energy absorbed in the high atmosphere (stratosphere), in the ozone (Herzberg continuum, 200-220 nm) and oxygen bands, and its high variability is most probably at the origin of a climate influence (UV affects stratospheric dynamics and temperatures, altering interplanetary waves and weather patterns both poleward and downward to the lower stratosphere and tropopause regions). Recent measurements at the time of the last solar minimum suggest that variations in the UV may be larger than previously assumed what implies a very different response in both stratospheric ozone and temperature. A simultaneous observation of the incoming FUV and of the ozone (O _{3}) production, would bring an invaluable information on this process of solar-climat forcing. Space instruments have already measured the different components of the Earth radiative budget but this is, to our knowledge, the first time that all instruments are operated on the same platform. This characteristic guarantees by itself obtaining original scientific results. SUMO is a 10x10x30 cm (3) nanosatellite (``3U"), the payload occupying ``1U", i.e. a cube of 10x10x10 cm (3) for 1 kg and 1 W of power. Orbit is polar since a further challenge in understanding the relation between solar UV variability and stratospheric ozone on arctic and antarctic regions. SUMO definition has been completed (platform and payload assembly integration and tests are possible in 24 months) and it is now intended to be proposed to CNES for a flight in 2017. Mission is expected to last up to 1 year. Follow-up is 2 fold: on

  17. Research progression on biologic function and effect of SUMO-specific proteinase%去SUMO化蛋白酶的生物学功能与作用基础研究

    Institute of Scientific and Technical Information of China (English)

    程金科; 左勇

    2012-01-01

    SUMO, an ubiquitin-like protein, can covalently conjugate proteins and plays a role in the regulation of target protein functions and localizations. Similar to ubiquitin, SUMO modification is a dynamic process catalyzed by El, E2? and E3 and reversed by Sentrin/SUMO-specific proteases ( SENPs) . To date, 6 SENP members have been defined in human cells, each of which has different cell localization and target specificity. However, the physiologic functions of SENPs are not fully understood. By analysis of SENPs knock-out mice, we find that SENP1 regulates hypoxia-HIFla signaling or androgen receptor ( AR) signaling through positive feedback loop, and plays an important role in HIFlot or AR-involved physiologic and pathologic processes. We also find that SENP2 is essential for suppression of polycomb group protein-mediated gene silencing during heart development. These results reveal the critical roles of SENPs in the regulation of targets-involved signaling,%小泛素样修饰蛋白(SUMO)修饰是一种新发现的类泛素蛋白质修饰形式.SUMO修饰由活化酶E1、接合酶E2和连接酶E3等三个酶相继作用来完成.同时它又是一个动态的、可逆的过程,其去SUMO修饰的过程则是由SUMO特异性蛋白酶(SENP)家族来介导.在蛋白质SUMO修饰的动态过程中,SENP介导的去SUMO过程是决定其靶蛋白质SUMO修饰水平的一个主要因素,同时也是SUMOylation被调节的一个主要环节.SENP家族包括SENP1、SENP2、SENP3、SENP5、SENP6、SENP7等成员,每个成员具有不同的细胞内定位和底物特异性.虽然对其生化特性进行了大量研究,但SENP在参与的细胞生命活动过程中的作用并未十分了解.我们通过分析SENP1和SENP2基因敲除小鼠的表型,发现了SENP1通过正反馈机制调控缺氧-HIF1α和雄激素受体所介导的信号通路,从而在HIF1α与AR所参与的生理病理过程中发挥了重要作用.同时,我们发现SENP2通过调控PeG的活性,参与心

  18. Expression and characterization of SUMO-conjugated metal-responsive transcription factor 1: SIM-dependent cross-interaction and distinct DNA binding activity.

    Science.gov (United States)

    Lin, Chang-Yi; Liu, Ya-Chuan; Lin, Meng-Chieh; Thi Nguyen, Thao; Tam, Ming F; Chein, Chih-Yuan; Lin, Meng-Ting; Lin, Lih-Yuan

    2013-04-01

    Metal-responsive transcription factor 1 (MTF-1) regulates a variety of genes involving in metal homeostasis and oxidative stress. We have shown that MTF-1 can be conjugated by small ubiquitin-like modifier (SUMO) and forms complexes with cellular factor(s) in a SUMO-interacting motif (SIM)-dependent manner. To investigate whether the interaction of MTF-1 and its SUMO conjugate occurs, we expressed and isolated MTF-1 and sumoylated MTF-1 (S-MTF-1) for functional studies. Various conditions were examined to optimize the expressions of MTF-1 and S-MTF-1. Results from affinity column chromatography demonstrated that the unmodified MTF-1 consistently co-eluted with the S-MTF-1. Mutations at the SIM did not reduce the level of MTF-1 sumoylation but the sumoylated product can then be purified to homogeneity. The presence of MTF-1 cross-interaction was further supported by in vitro pull-down assays. The ability of the purified proteins in binding metal-responsive element (MRE) was assessed with electrophoretic mobility shift assay. Noticeably, MTF-1 required the presence of cell extracts to render the binding activity. However, S-MTF-1 binds MRE in void of other cellular factors. The same characteristic was found for MTF-1 with SUMO fusion at the carboxyl terminus. These results indicate that the presence of SUMO moiety allows the protein to interact directly with MRE.

  19. Mutation in SUMO E3 ligase, SIZ1, disrupts the mature female gametophyte in Arabidopsis

    KAUST Repository

    Ling, Yu

    2012-01-09

    Female gametophyte is the multicellular haploid structure that can produce embryo and endosperm after fertilization, which has become an attractive model system for investigating molecular mechanisms in nuclei migration, cell specification, cell-to-cell communication and many other processes. Previous reports found that the small ubiquitin-like modifier (SUMO) E3 ligase, SIZ1, participated in many processes depending on particular target substrates and suppression of salicylic acid (SA) accumulation. Here, we report that SIZ1 mediates the reproductive process. SIZ1 showed enhanced expression in female organs, but was not detected in the anther or pollen. A defect in the siz1-2 maternal source resulted in reduced seed-set regardless of high SA concentration within the plant. Moreover, aniline blue staining and scanning electron microscopy revealed that funicular and micropylar pollen tube guidance was arrested in siz1-2 plants. Some of the embryo sacs of ovules in siz1-2 were also disrupted quickly after stage FG7. There was no significant affects of the siz1-2 mutation on expression of genes involved in female gametophyte development- or pollen tube guidance in ovaries. Together, our results suggest that SIZ1 sustains the stability and normal function of the mature female gametophyte which is necessary for pollen tube guidance. © 2012 Ling et al.

  20. Ubiquitin modifications

    OpenAIRE

    Swatek, Kirby N.; Komander, David

    2016-01-01

    Protein ubiquitination is a dynamic multifaceted post-translational modification involved in nearly all aspects of eukaryotic biology. Once attached to a substrate, the 76-amino acid protein ubiquitin is subjected to further modifications, creating a multitude of distinct signals with distinct cellular outcomes, referred to as the 'ubiquitin code'. Ubiquitin can be ubiquitinated on seven lysine (Lys) residues or on the N-terminus, leading to polyubiquitin chains that can encompass complex top...

  1. Functional characterization of the SIZ/PIAS-type SUMO E3 ligases, OsSIZ1 and OsSIZ2 in rice

    KAUST Repository

    Park, Hyeongcheol

    2010-06-18

    Sumoylation is a post-translational regulatory process in diverse cellular processes in eukaryotes, involving conjugation/deconjugation of small ubiquitin-like modifier (SUMO) proteins to other proteins thus modifying their function. The PIAS [protein inhibitor of activated signal transducers and activators of transcription (STAT)] and SAP (scaffold attachment factor A/B/acinus/PIAS)/MIZ (SIZ) proteins exhibit SUMO E3-ligase activity that facilitates the conjugation of SUMO proteins to target substrates. Here, we report the isolation and molecular characterization of Oryza sativa SIZ1 (OsSIZ1) and SIZ2 (OsSIZ2), rice homologs of Arabidopsis SIZ1. The rice SIZ proteins are localized to the nucleus and showed sumoylation activities in a tobacco system. Our analysis showed increased amounts of SUMO conjugates associated with environmental stresses such as high and low temperature, NaCl and abscisic acid (ABA) in rice plants. The expression of OsSIZ1 and OsSIZ2 in siz1-2 Arabidopsis plants partially complemented the morphological mutant phenotype and enhanced levels of SUMO conjugates under heat shock conditions. In addition, ABA-hypersensitivity of siz1-2 seed germination was partially suppressed by OsSIZ1 and OsSIZ2. The results suggest that rice SIZ1 and SIZ2 are able to functionally complement Arabidopsis SIZ1 in the SUMO conjugation pathway. Their effects on the Arabidopsis mutant suggest a function for these genes related to stress responses and stress adaptation. © 2010 Blackwell Publishing Ltd.

  2. Intranuclear targeting and nuclear export of the adenovirus E1B-55K protein are regulated by SUMO1 conjugation

    OpenAIRE

    Kindsmüller, Kathrin; Groitl, Peter; Härtl, Barbara; Blanchette, Paola; Hauber, Joachim; Dobner, Thomas

    2007-01-01

    We have investigated the requirements for CRM1-mediated nuclear export and SUMO1 conjugation of the adenovirus E1B-55K protein during productive infection. Our data show that CRM1 is the major export receptor for E1B-55K in infected cells. Functional inactivation of the E1B-55K CRM1-dependent nuclear export signal (NES) or leptomycin B treatment causes an almost complete redistribution of the viral protein from the cytoplasm to the nucleus and its accumulation at the periphery of the viral re...

  3. 8051 Mikrodenetleyicili Bir Sumo Robot Tasarımı ve Uygulaması

    Directory of Open Access Journals (Sweden)

    Mehmet ALBAYRAK

    2009-04-01

    Full Text Available Bu çalısmada, 8051 mikrodenetleyici kontrollü bir sumo robotun mekanik tasarımı, kontrol kartı ve yazılımı gerçeklestirilmistir. Robot, yarısma ve egitim amaçlı tasarlandıgı için hücum amaçlı strateji içeren yazılım gelistirilerek yüklenmistir. 8051 mikrodenetleyici içindeki kontrol yazılımı PC 'nin seri portuna baglı mikrodenetleyici programlama kartına takılıp, kolaylıkla güncellenebildigi için yazılım stratejisi her yarısma için degistirilebilir niteliktedir. Microdenetleyici yazılımı μVision Keil derleyicisinde C kodu kullanılarak hazırlanmıstır. Mekanik tasarım asamasında robot dıs yüzeyinin darbelere dayanıklı malzemeden seçilmesine ve tekerleklerin sürtünmesinin fazla (kaymaz olmasına özen gösterilmistir. Robotun gücünü belirlenen agırlık sınırları içinde en fazla hale getirebilmek için 4 ayrı disli DC motor ve 4 amperlik akü kullanılmıstır. Rakip algılama mesafe sensörü ve kontrast sensörü (beyaz algılama seçiminde cevap verme süresi çok az olan sensörler tercih edilmistir.

  4. SUMO ligase PIAS1 functions as a target gene selective androgen receptor coregulator on prostate cancer cell chromatin.

    Science.gov (United States)

    Toropainen, Sari; Malinen, Marjo; Kaikkonen, Sanna; Rytinki, Miia; Jääskeläinen, Tiina; Sahu, Biswajyoti; Jänne, Olli A; Palvimo, Jorma J

    2015-01-01

    Androgen receptor (AR) is a ligand-activated transcription factor that plays a central role in the development and growth of prostate carcinoma. PIAS1 is an AR- and SUMO-interacting protein and a putative transcriptional coregulator overexpressed in prostate cancer. To study the importance of PIAS1 for the androgen-regulated transcriptome of VCaP prostate cancer cells, we silenced its expression by RNAi. Transcriptome analyses revealed that a subset of the AR-regulated genes is significantly influenced, either activated or repressed, by PIAS1 depletion. Interestingly, PIAS1 depletion also exposed a new set of genes to androgen regulation, suggesting that PIAS1 can mask distinct genomic loci from AR access. In keeping with gene expression data, silencing of PIAS1 attenuated VCaP cell proliferation. ChIP-seq analyses showed that PIAS1 interacts with AR at chromatin sites harboring also SUMO2/3 and surrounded by H3K4me2; androgen exposure increased the number of PIAS1-occupying sites, resulting in nearly complete overlap with AR chromatin binding events. PIAS1 interacted also with the pioneer factor FOXA1. Of note, PIAS1 depletion affected AR chromatin occupancy at binding sites enriched for HOXD13 and GATA motifs. Taken together, PIAS1 is a genuine chromatin-bound AR coregulator that functions in a target gene selective fashion to regulate prostate cancer cell growth. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. SUMO proteases ULP1c and ULP1d are required for development and osmotic stress responses in Arabidopsis thaliana

    DEFF Research Database (Denmark)

    Castro, Pedro Humberto Araújo R F; Couto, Daniel; Freitas, Sara

    2016-01-01

    diminished in vitro root growth under low water potential and higher stomatal aperture, yet leaf transpirational water loss and whole drought tolerance were not significantly altered. Generation of a triple siz1 ulp1c ulp1d mutant suggests that ULP1c/d and the SUMO E3 ligase SIZ1 may display separate...

  6. Study of the role of functional variants of SLC22A4, RUNX1 and SUMO4 in systemic lupus erythematosus

    Science.gov (United States)

    Orozco, G; Sánchez, E; Gómez, L M; González‐Gay, M A; López‐Nevot, M A; Torres, B; Ortego‐Centeno, N; Jiménez‐Alonso, J; de Ramón, E; Román, J Sánchez; Anaya, J M; Sturfelt, G; Gunnarsson, I; Svennungsson, E; Alarcón‐Riquelme, M; González‐Escribano, M F; Martín, J

    2006-01-01

    Background Functional polymorphisms of the solute carrier family 22, member 4 (SLC22A4), runt related transcription factor 1 (RUNX1) and small ubiquitin‐like modifier 4 (SUMO4) genes have been shown to be associated with several autoimmune diseases. Objective To test the possible role of these variants in susceptibility to or severity of systemic lupus erythematosus (SLE), on the basis that common genetic bases are shared by autoimmune disorders. Methods 597 SLE patients and 987 healthy controls of white Spanish origin were studied. Two additional cohorts of 228 SLE patients from Sweden and 122 SLE patients from Colombia were included. A case–control association study was carried out with six single nucleotide polymorphisms (SNP) spanning the SLC22A4 gene, one SNP in RUNX1 gene, and one additional SNP in SUM04 gene. Results No significant differences were observed between SLE patients and healthy controls when comparing the distribution of the genotypes or alleles of any of the SLC22A4, RUNX1, or SUMO4 polymorphisms tested. Significant differences were found in the distribution of the SUMO4 genotypes and alleles among SLE patients with and without nephritis, but after multiple testing correction, the significance of the association was lost. The association of SUMO4 with nephritis could not be verified in two independent SLE cohorts from Sweden and Colombia. Conclusions These results suggest that the SLC22A4, RUNX1, and SUMO4 polymorphisms analysed do not play a role in the susceptibility to or severity of SLE. PMID:16249223

  7. SUMO4基因多态性与NF-κB、糖尿病及冠心病的关系%Role of SUMO4 gene polymorphism in NF-κB signalling pathway and its research advances in genetic susceptibility to DM and CAD

    Institute of Scientific and Technical Information of China (English)

    蒲连美; 南楠; 杨泽; 金泽宁; 刘铭

    2012-01-01

    Diabetes mellitus ( DM ) and coronary artery disease ( CAD ) are both complex chronic diseases which are caused by interactions of multiple genes with environmental factors.Nuclear factor kappa- B ( NF-κB ) is a family of critical transcription factors modulating the expression of diverse genes involved in the pathogenesis of several inflammatory diseases,including DM and CAD. IκBα is one of the most important inhibitory proteins for NF-κB.SUMO4 conjugates IκBα and stabilizes it from degradation.As a result NF-κB is kept inactive in the cytosol by forming a complex with IκBα and SUMO4.Moreover dissociated NF-κB binds to SUMO4 promoter and activates its transcription.S(o) SUMO4 can function as a negative feedback regulator for the NF-κB signaling pathway.Studies confirmed that SUMO4 M55V substitution results in significant higher levels of NF-κB activity and is associated with susceptibility to DM and CAD.This review focuses on the role of SUMO4 gene polymorphism in NF-κB signalling pathway and its research advances in genetic susceptibility to DM and CAD.%糖尿病( diabetes mellitus,DM)和冠心病(coronary artery disease,CAD)均为多基因介导、基因和环境相互作用的慢性复杂疾病.核因子κB(nuclear factor kappa B,NF- κB)是一类转录因子家族,在细胞内信号传导系统中起关键作用,参与包括DM和CAD在内的多种炎症性疾病的发生发展.研究证实小泛素样修饰蛋白4( small ubiquitin- like modi - fier 4,SUMO4)使NF- κB的抑制剂IκBα免于降解,负性调节NF- κB的转录活性,另外游离的NF- κB能够与SUMO4基因启动子结合并激活SUMO4基因的转录.而SUMO4基因多态性导致NF-κB活性显著增加,并与DM和CAD的易感性相关.此文主要就SUMO4基因多态性对NF-κB转录活性的影响及其在DM和CAD遗传易感性的关联研究进展作一概述.

  8. Expressions of SUMO4, NF-κB and IκB in kidney of rats with type 2diabetes mellitus and its significance%SUMO4、NF-κB、IκB在2型糖尿病大鼠肾脏中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    陈思娇; 王大南; 徐锦春; 李红燕; 刘芙蓉; 宋今丹

    2012-01-01

    Objective:To study expressions of small ubiquitin-related modifier protein (SUMO) 4 (SUMO4),nuclear factor (NF) -κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM).Methods:A total of ten 40-week-old male Goto-Kakizaki (GK) rats (with spontaneous diabetes mellitus) of specificpathogen free (SPF) grade,and ten 40-week-old male Wistar rats of SPF grade were selected.The lesion of renal tissue was observed by hematoxylin eosin (HE) staining.Experssions of SUMO4,NF-κB and IκB in renal tissue were observed by immunohistochemistry methods.Results:In the GK rats,glomerular capillary ball hypertrophy,basilar membrane slightly thickening; glomerular mesangial cells hyperplasia,hypertrophy and renal tubular epithelial cells hypertrophy were observed.Compared with normal Wistar rats,expression levels of NF-κB [ (0.232 ±0.034) vs.(0.634± 0.058)],IκB [ (0.242 ± 0.027) vs.(0.712 ± 0.078)] and SUMO4 [ (0.160 ± 0.031) vs.(0.545 ± 0.045)] significantly increased in renal tissue of GK rats (P<0.01 all).Conclusion:Compared with Wistar rats,expressions of NF-κB,IκB and SUMO4 significantly increase in renal tissue of GK rats,suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats.Therefore,sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.%目的:探讨2型糖尿病大鼠肾脏组织小泛素相关修饰蛋白4 (SUMO4)、核转录因子(NF) -κB、NF-κB的抑制因子(IκB)的表达及意义.方法:取10只40周龄的无特定病原体(SPF)级雄性自发性糖尿病(GK)大鼠,10只40周龄的SPF级雄性Wistar大鼠,通过HE染色法观察肾组织病变、免疫组化法观察肾组织的SUMO4与IκB、SUMO4与NF-κB表达情况.结果:GK大鼠的肾小球毛细血管球肥大,基底膜轻度增厚,肾小球系膜细胞增生、肥大,肾小管上皮细胞肥大.与正常Wistar大鼠比较,GK大鼠的肾脏NF-κB[ (0.232±0

  9. Posttranslational Modifications of the Master Transcriptional Regulator NPR1 Enable Dynamic but Tight Control of Plant Immune Responses

    Science.gov (United States)

    Saleh, Abdelaty; Withers, John; Mohan, Rajinikanth; Marqués, Jorge; Gu, Yangnan; Yan, Shunping; Zavaliev, Raul; Nomoto, Mika; Tada, Yasuomi; Dong, Xinnian

    2015-01-01

    Summary NPR1, a master regulator of basal and systemic acquired resistance in plants, confers immunity through a transcriptional cascade, which includes transcription activators (e.g., TGA3) and repressors (e.g., WRKY70), leading to the massive induction of antimicrobial genes. How this single protein orchestrates genome-wide transcriptional reprogramming in response to immune stimulus remains a major question. Paradoxically, while NPR1 is essential for defense gene induction, its turnover appears to be required for this function, suggesting that NPR1 activity and degradation are dynamically regulated. Here we show that sumoylation of NPR1 by SUMO3 activates defense gene expression by switching NPR1's association with the WRKY transcription repressors to TGA transcription activators. Sumoylation also triggers NPR1 degradation, rendering the immune induction transient. SUMO modification of NPR1 is inhibited by phosphorylation at Ser55/Ser59, which keeps NPR1 stable and quiescent. Thus, posttranslational modifications enable dynamic but tight and precise control of plant immune responses. PMID:26269953

  10. Small Ubiquitin-Like Modifier Conjugating Enzyme with Active Site Mutation Acts as Dominant Negative Inhibitor of SUMO Conjugation in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Konstantin Tomanov; Christian Hardtke; Ruchika Budhiraja; Rebecca Hermkes; George Coupland; Andreas Bachmair

    2013-01-01

    Small ubiquitin-like modifier (SUMO) conjugation affects a broad range of processes in plants,including growth,flower initiation,pathogen defense,and responses to abiotic stress.Here,we investigate in vivo and in vitro a SUMO conjugating enzyme with a Cys to Ser change in the active site,and show that it has a dominant negative effect.In planta expression significantly perturbs normal development,leading to growth retardation,early flowering and gene expression changes.We suggest that the mutant protein can serve as a probe to investigate sumoylation,also in plants for which poor genetic infrastructure precludes analysis via loss-of-function mutants.

  11. High-level SUMO-mediated fusion expression of ABP-dHC-cecropin A from multiple joined genes in Escherichia coli.

    Science.gov (United States)

    Zhang, Jiaxin; Movahedi, Ali; Wei, Zhiheng; Sang, Ming; Wu, Xiaolong; Wang, Mengyang; Wei, Hui; Pan, Huixin; Yin, Tongming; Zhuge, Qiang

    2016-09-15

    The antimicrobial peptide ABP-dHC-cecropin A is a small cationic peptide with potent activity against a wide range of bacterial species. Evidence of antifungal activity has also been suggested; however, evaluation of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve the expression level of ABP-dHC-cecropin A in E. coli, tandem repeats of the ABP-dHC-cecropin A gene were constructed and expressed as fusion proteins (SUMO-nABP-dHC-cecropin, n = 1, 2, 3, 4) via pSUMO-nABP-dHC-cecropin A vectors (n = 1, 2, 3, 4). Comparison of the expression levels of soluble SUMO-nABP-dHC-cecropin A fusion proteins (n = 1, 2, 3, 4) suggested that BL21 (DE3)/pSUMO-3ABP-dHC-cecropin A is an ideal recombinant strain for ABP-dHC-cecropin A production. Under the selected conditions of cultivation and isopropylthiogalactoside (IPTG) induction, the expression level of ABP-dHC-cecropin A was as high as 65 mg/L, with ∼21.3% of the fusion protein in soluble form. By large-scale fermentation, protein production reached nearly 300 mg/L, which is the highest yield of ABP-dHC-cecropin A reported to date. In antibacterial experiments, the efficacy was approximately the same as that of synthetic ABP-dHC-cecropin A. This method provides a novel and effective means of producing large amounts of ABP-dHC-cecropin A.

  12. Recombinant Scorpine Produced Using SUMO Fusion Partner in Escherichia coli Has the Activities against Clinically Isolated Bacteria and Inhibits the Plasmodium falciparum Parasitemia In Vitro

    OpenAIRE

    Chao Zhang; Xinlong He; Yaping Gu; Huayun Zhou; Jun Cao; Qi Gao

    2014-01-01

    Scorpine, a small cationic peptide from the venom of Pandinus imperator, which has been shown to have anti-bacterial and anti-plasmodial activities, has potential important applications in the pharmaceutical industries. However, the isolation of scorpine from natural sources is inefficient and time-consuming. Here, we first report the expression and purification of recombinant scorpine in Escherichia coli, using small ubiquitin-related modifier (SUMO) fusion partner. The fusion protein was ex...

  13. Role of the SUMO-interacting motif in HIPK2 targeting to the PML nuclear bodies and regulation of p53

    Energy Technology Data Exchange (ETDEWEB)

    Sung, Ki Sa; Lee, Yun-Ah [Department of Biological Sciences, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of); Kim, Eui Tae [Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon 440-746 (Korea, Republic of); Lee, Seung-Rock [Department of Biochemistry, Research Center for Aging and Geriatrics, Research Institute of Medical Sciences, Chonnam National University Medical School, Gwangju 501-190 (Korea, Republic of); Ahn, Jin-Hyun [Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Suwon 440-746 (Korea, Republic of); Choi, Cheol Yong, E-mail: choicy@skku.ac.kr [Department of Biological Sciences, Sungkyunkwan University, Suwon 440-746 (Korea, Republic of)

    2011-04-15

    Homeodomain-interacting protein kinase 2 (HIPK2) is a key regulator of various transcription factors including p53 and CtBP in the DNA damage signaling pathway. PML-nuclear body (NB) is required for HIPK2-mediated p53 phosphorylation at Ser46 and induction of apoptosis. Although PML-NB targeting of HIPK2 has been shown, much is not clear about the molecular mechanism of HIPK2 recruitment to PML-NBs. Here we show that HIPK2 colocalizes specifically with PML-I and PML-IV. Mutational analysis showed that HIPK2 recruitment to PML-IV-NBs is mediated by the SUMO-interaction motifs (SIMs) of both PML-IV and HIPK2. Wild-type HIPK2 associated with SUMO-conjugated PML-IV at a higher affinity than with un-conjugated PML-IV, while the association of a HIPK2 SIM mutant with SUMO-modified PML-IV was impaired. In colony formation assays, HIPK2 strongly suppressed cell proliferation, but HIPK2 SIM mutants did not. In addition, activation and phosphorylation of p53 at the Ser46 residue were impaired by HIPK2 SIM mutants. These results suggest that SIM-mediated HIPK2 targeting to PML-NBs is crucial for HIPK2-mediated p53 activation and induction of apoptosis.

  14. Recombinant scorpine produced using SUMO fusion partner in Escherichia coli has the activities against clinically isolated bacteria and inhibits the Plasmodium falciparum parasitemia in vitro.

    Directory of Open Access Journals (Sweden)

    Chao Zhang

    Full Text Available Scorpine, a small cationic peptide from the venom of Pandinus imperator, which has been shown to have anti-bacterial and anti-plasmodial activities, has potential important applications in the pharmaceutical industries. However, the isolation of scorpine from natural sources is inefficient and time-consuming. Here, we first report the expression and purification of recombinant scorpine in Escherichia coli, using small ubiquitin-related modifier (SUMO fusion partner. The fusion protein was expressed in soluble form in E. coli, and expression was verified by SDS-PAGE and western blotting analysis. The fusion protein was purified to 90% purity by nickel-nitrilotriacetic acid (Ni2+-NTA resin chromatography. After the SUMO-scorpine fusion protein was cleaved by the SUMO protease, the cleaved sample was reapplied to a Ni2+-NTA column. Tricine/SDS-PAGE gel results indicated that Scorpine had been purified successfully to more than 95% purity. The recombinantly expressed Scorpine showed anti-bacterial activity against two standard bacteria including Staphylococcus aureus ATCC 29213 and Acinetobacter baumannii ATCC 19606, and clinically isolated bacteria including S. aureus S, S. aureus R, A. baumannii S, and A. baumannii R. It also produced 100% reduction in Plasmodium falciparum parasitemia in vitro. Thus, the expression strategy presented in this study allowed convenient high yield and easy purification of recombinant Scorpine for pharmaceutical applications in the future.

  15. Recombinant scorpine produced using SUMO fusion partner in Escherichia coli has the activities against clinically isolated bacteria and inhibits the Plasmodium falciparum parasitemia in vitro.

    Science.gov (United States)

    Zhang, Chao; He, Xinlong; Gu, Yaping; Zhou, Huayun; Cao, Jun; Gao, Qi

    2014-01-01

    Scorpine, a small cationic peptide from the venom of Pandinus imperator, which has been shown to have anti-bacterial and anti-plasmodial activities, has potential important applications in the pharmaceutical industries. However, the isolation of scorpine from natural sources is inefficient and time-consuming. Here, we first report the expression and purification of recombinant scorpine in Escherichia coli, using small ubiquitin-related modifier (SUMO) fusion partner. The fusion protein was expressed in soluble form in E. coli, and expression was verified by SDS-PAGE and western blotting analysis. The fusion protein was purified to 90% purity by nickel-nitrilotriacetic acid (Ni2+-NTA) resin chromatography. After the SUMO-scorpine fusion protein was cleaved by the SUMO protease, the cleaved sample was reapplied to a Ni2+-NTA column. Tricine/SDS-PAGE gel results indicated that Scorpine had been purified successfully to more than 95% purity. The recombinantly expressed Scorpine showed anti-bacterial activity against two standard bacteria including Staphylococcus aureus ATCC 29213 and Acinetobacter baumannii ATCC 19606, and clinically isolated bacteria including S. aureus S, S. aureus R, A. baumannii S, and A. baumannii R. It also produced 100% reduction in Plasmodium falciparum parasitemia in vitro. Thus, the expression strategy presented in this study allowed convenient high yield and easy purification of recombinant Scorpine for pharmaceutical applications in the future.

  16. Studi Kinerja VANET Scenario Generators: SUMO dan VanetMobisim untuk Implementasi Routing Protocol AODV menggunakan Network Simulator 2 (NS-2

    Directory of Open Access Journals (Sweden)

    Firdaus Nutrihadi

    2016-04-01

    Full Text Available Vehicular Ad Hoc Network (VANET merupakan turunan dari MANET (Mobile Ad Hoc Network sebagai inovasi baru dalam dunia teknologi yang membantu kebutuhan manusia dalam berkomunikasi. VANET dapat mendukung komunikasi langsung antara kendaraan (Vehicle to Vehicle dan antara kendaraan-infrastruktur (Vehicle to Infrastructure dengan adanya infrastruktur jaringan nirkabel. Namun, implementasi VANET di dunia masih sulit dilakukan sehingga banyak penelitian dilakukan dengan membuat simulasi menggunakan mobility generator dan network simulator. Pada makalah ini yang diteliti yaitu performa skema VANET yang dihasilkan oleh mobility generator SUMO dan Vanetmobisim. Penelitian ini menggunakan NS-2 sebagai simulator VANET dengan protokol reaktif AODV sebagai routing protocol. Skenario VANET dengan peta berbentuk grid dan peta riil Sutomo,Surabaya digunakan pada kedua generator SUMO dan VanetMobisim dengan memvariasikan jumlah kendaraan simulasi. Matriks evaluasi kinerja yang digunakan dalam penelitian ini adalah packet delivery ratio, end-to-end delay, dan routing overhead. Dalam ketiga skenario, performa routing protokol SUMO-AODV lebih baik. VanetMobisim-AODV, dikarenakan lebih banyak lalu lintas dan rute yang putus, menghasilkan performa yang baik namun masih di bawah pesaingnya.

  17. Readers of histone modifications

    Institute of Scientific and Technical Information of China (English)

    Miyong Yun; Jun Wu; Jerry L Workman; Bing Li

    2011-01-01

    Histone modifications not only play important roles in regulating chromatin structure and nuclear processes but also can be passed to daughter cells as epigenetic marks.Accumulating evidence suggests that the key function of histone modifications is to signal for recruitment or activity of downstream effectors. Here, we discuss the latest discovery of histone-modification readers and how the modification language is interpreted.

  18. Production of aggregation prone human interferon gamma and its mutant in highly soluble and biologically active form by SUMO fusion technology.

    Science.gov (United States)

    Tileva, M; Krachmarova, E; Ivanov, I; Maskos, K; Nacheva, G

    2016-01-01

    The Escherichia coli expression system is a preferable choice for production of recombinant proteins. A disadvantage of this system is the target protein aggregation in "inclusion bodies" (IBs) that further requires solubilisation and refolding, which is crucial for the properties and the yield of the final product. In order to prevent aggregation, SUMO fusion tag technology has been successfully applied for expression of eukaryotic proteins, including human interferon gamma (hIFNγ) that was reported, however, with no satisfactory biological activity. We modified this methodology for expression and purification of both the wild type hIFNγ and an extremely prone to aggregation mutant hIFNγ-K88Q, whose recovery from IBs showed to be ineffective upon numerous conditions. By expression of the N-terminal His-SUMO fusion proteins in the E. coli strain BL21(DE3)pG-KJE8, co-expressing two chaperone systems, at 24 °C a significant increase in solubility of both target proteins (1.5-fold for hIFNγ and 8-fold for K88Q) was achieved. Two-step chromatography (affinity and ion-exchange) with on-dialysis His-SUMO-tag cleavage was applied for protein purification that yielded 6.0-7.0mg/g wet biomass for both proteins with >95% purity and native N-termini. The optimised protocol led to increased yields from 5.5 times for hIFNγ up to 100 times for K88Q in comparison to their isolation from IBs. Purified hIFNγ showed preserved thermal stability and antiproliferative activity corresponding to that of the native reference sample (3 × 10(7)IU/mg). The developed methodology represents an optimised procedure that can be successfully applied for large scale expression and purification of aggregation-prone proteins in soluble native form.

  19. La tradición oral de los indígenas Sumos: Características y temáticas

    OpenAIRE

    Arguedas, Gilda Rosa

    2015-01-01

    Este artículo se ocupa de la tradición oral de los "Sumo" indios, uno de los grupos que habitan la costa atlántica de Honduras y Nicaragua, también conocido como "Mosquitia".Las historias analizadas (36) son principalmente narraciones aunque hay algunas descripciones y explicaciones. De acuerdo con el momento en el que se establece la narración, estas historias se clasifican en tres grupos: los mitos, leyendas y cuentos. Los temas de las historias son variadas, aunque los elementos míticos pr...

  20. Triptolide inhibits the proliferation of prostate cancer cells and down-regulates SUMO-specific protease 1 expression.

    Directory of Open Access Journals (Sweden)

    Weiwei Huang

    Full Text Available Recently, traditional Chinese medicine and medicinal herbs have attracted more attentions worldwide for its anti-tumor efficacy. Celastrol and Triptolide, two active components extracted from the Chinese herb Tripterygium wilfordii Hook F (known as Lei Gong Teng or Thunder of God Vine, have shown anti-tumor effects. Celastrol was identified as a natural 26 s proteasome inhibitor which promotes cell apoptosis and inhibits tumor growth. The effect and mechanism of Triptolide on prostate cancer (PCa is not well studied. Here we demonstrated that Triptolide, more potent than Celastrol, inhibited cell growth and induced cell death in LNCaP and PC-3 cell lines. Triptolide also significantly inhibited the xenografted PC-3 tumor growth in nude mice. Moreover, Triptolide induced PCa cell apoptosis through caspases activation and PARP cleavage. Unbalance between SUMOylation and deSUMOylation was reported to play an important role in PCa progression. SUMO-specific protease 1 (SENP1 was thought to be a potential marker and therapeutical target of PCa. Importantly, we observed that Triptolide down-regulated SENP1 expression in both mRNA and protein levels in dose-dependent and time-dependent manners, resulting in an enhanced cellular SUMOylation in PCa cells. Meanwhile, Triptolide decreased AR and c-Jun expression at similar manners, and suppressed AR and c-Jun transcription activity. Furthermore, knockdown or ectopic SENP1, c-Jun and AR expression in PCa cells inhibited the Triptolide anti-PCa effects. Taken together, our data suggest that Triptolide is a natural compound with potential therapeutic value for PCa. Its anti-tumor activity may be attributed to mechanisms involving down-regulation of SENP1 that restores SUMOylation and deSUMOyaltion balance and negative regulation of AR and c-Jun expression that inhibits the AR and c-Jun mediated transcription in PCa.

  1. Behavior Modification in Coaching.

    Science.gov (United States)

    Lynch, Annette Rutt; Stillman, Stephen M.

    1979-01-01

    An example of behavior modification used in athletic coaching is presented. The case study involves a member of a women's basketball team and details the use of behavior modification for both weight reduction and skill improvement. (JMF)

  2. Detection of O-propargyl-puromycin with SUMO and ubiquitin by click chemistry at PML-nuclear bodies during abortive proteasome activities

    Energy Technology Data Exchange (ETDEWEB)

    Uozumi, Naoki; Matsumoto, Hotaru [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Saitoh, Hisato, E-mail: hisa@kumamoto-u.ac.jp [Course for Biological Sciences, Faculty of Science, Kumamoto University, Kumamoto (Japan); Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kumamoto (Japan)

    2016-05-27

    The amino-nucleoside antibiotic, puromycin, acts by covalently linking to elongating polypeptide chains on ribosomes to generate prematurely terminated immature polypeptides. The trafficking of puromycin-conjugated (puromycylated) immature polypeptides within cell has, however, remained elusive. In this study, using O-propargyl-puromycin (OP-Puro), the distribution of puromycylated polypeptides was assessed in HeLa cells by click chemistry. Under standard culture conditions, OP-Puro signals were detected in the cytoplasm and nucleus with the highest concentrations in the nucleolus. Intriguingly, when proteasome activities were aborted using MG132, OP-Puro signals began to accumulate at promyelocytic leukemia nuclear bodies (PML-NBs) in addition to the nucleolus. We also found promiscuous association of OP-Puro signals with SUMO-2/3 and ubiquitin at PML-NBs, but not at the nucleolus, during abortive proteasome activities. This study reveals a previously unknown distribution of OP-Puro that argues for a nuclear function in regulating immature protein homeostasis. -- Highlights: •Click chemistry detects O-propargyl-puromycin (OP-Puro) signals in the nucleus. •OP-Puro accumulates at PML-NBs during abortive proteasome activities. •SUMO and ubiquitin are promiscuously associated with OP-Puro at PML-NBs. •The nucleus may function in immature protein homeostasis.

  3. Effects of Nephrolithiasis on Serum DNase (Deoxyribonuclease I and II) Activity and E3 SUMO-Protein Ligase NSE2 (NSMCE2) in Malaysian Individuals

    Institute of Scientific and Technical Information of China (English)

    Faridah Yusof; Atheer Awad Mehde; Wesen Adel Mehdi; Raha Ahmed Raus; Hamid Ghazali; Azlina Abd Rahman

    2015-01-01

    Objective Nephrolithiasis is one of the most common disorders of the urinary tract. The aim of this study was to examine a possible relationship between DNase I/II activity and E3 SUMO-protein ligase NSE2 in the sera of nephrolithiasis patients to evaluate the possibility of a new biomarker for evaluating kidney damage. Methods Sixty nephrolithiasis patients and 50 control patients were enrolled in a case-control study. Their blood urea, creatinine, protein levels and DNase I/II activity levels were measured by spectrometry. Serum NSMCE2 levels were measured by ELISA. Blood was collected from patients of the government health clinics in Kuantan-Pahang and fulfilled the inclusion criteria. Results The result indicated that mean levels of sera NSMCE2 have a significantly increase (P Conclusion This study suggests that an increase in serum concentrations of DNase I/II and E3 SUMO-protein ligase NSE2 level can be used as indicators for the diagnosis of kidney injury in patients with nephrolithiasis.

  4. Sgs1's roles in DNA end resection, HJ dissolution, and crossover suppression require a two-step SUMO regulation dependent on Smc5/6.

    Science.gov (United States)

    Bermúdez-López, Marcelino; Villoria, María Teresa; Esteras, Miguel; Jarmuz, Adam; Torres-Rosell, Jordi; Clemente-Blanco, Andres; Aragon, Luis

    2016-06-01

    The RecQ helicase Sgs1 plays critical roles during DNA repair by homologous recombination, from end resection to Holliday junction (HJ) dissolution. Sgs1 has both pro- and anti-recombinogenic roles, and therefore its activity must be tightly regulated. However, the controls involved in recruitment and activation of Sgs1 at damaged sites are unknown. Here we show a two-step role for Smc5/6 in recruiting and activating Sgs1 through SUMOylation. First, auto-SUMOylation of Smc5/6 subunits leads to recruitment of Sgs1 as part of the STR (Sgs1-Top3-Rmi1) complex, mediated by two SUMO-interacting motifs (SIMs) on Sgs1 that specifically recognize SUMOylated Smc5/6. Second, Smc5/6-dependent SUMOylation of Sgs1 and Top3 is required for the efficient function of STR. Sgs1 mutants impaired in recognition of SUMOylated Smc5/6 (sgs1-SIMΔ) or SUMO-dead alleles (sgs1-KR) exhibit unprocessed HJs at damaged replication forks, increased crossover frequencies during double-strand break repair, and severe impairment in DNA end resection. Smc5/6 is a key regulator of Sgs1's recombination functions.

  5. Effects of Nephrolithiasis on Serum DNase (Deoxyribonuclease I and II) Activity and E3 SUMO-Protein Ligase NSE2 (NSMCE2) in Malaysian Individuals.

    Science.gov (United States)

    Yusof, Faridah; Mehde, Atheer Awad; Mehdi, Wesen Adel; Raus, Raha Ahmed; Ghazali, Hamid; Rahman, Azlina Abd

    2015-09-01

    Nephrolithiasis is one of the most common disorders of the urinary tract. The aim of this study was to examine a possible relationship between DNase I/II activity and E3 SUMO-protein ligase NSE2 in the sera of nephrolithiasis patients to evaluate the possibility of a new biomarker for evaluating kidney damage. Sixty nephrolithiasis patients and 50 control patients were enrolled in a case-control study. Their blood urea, creatinine, protein levels and DNase I/II activity levels were measured by spectrometry. Serum NSMCE2 levels were measured by ELISA. Blood was collected from patients of the government health clinics in Kuantan-Pahang and fulfilled the inclusion criteria. The result indicated that mean levels of sera NSMCE2 have a significantly increase (P<0.01) in patients compared to control group. Compared with control subjects, activities and specific activities of serum DNase I and II were significantly elevated in nephrolithiasis patients (P$lt;0.01). This study suggests that an increase in serum concentrations of DNase I/II and E3 SUMO-protein ligase NSE2 level can be used as indicators for the diagnosis of kidney injury in patients with nephrolithiasis. Copyright © 2015 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  6. Enzymatic modification of starch

    DEFF Research Database (Denmark)

    Jensen, Susanne Langgård

    In the food industry approaches for using bioengineering are investigated as alternatives to conventional chemical and physical starch modification techniques in development of starches with specific properties. Enzyme-assisted post-harvest modification is an interesting approach to this, since...... branching connecting larger chain segments. In case of high BE activity this transfer happened prior to hydration and phase separation. The starch substrates thereby became locked in their granular structure and blocked furher access of BE. Transferase-based modification of starch has today almost...... exclusively been conducted on gelatinized starch. This study provides a new concept for transferase-based modification of starches in granular state....

  7. Licensing by modification Licensing by modification

    Directory of Open Access Journals (Sweden)

    Veneeta Dayal

    2008-04-01

    Full Text Available Abstract: This paper looks at the phenomena of licensing by modification, whereby an unacceptable noun phrase is redeemed by the presence of a modifier or an unavailable reading for a noun phrase is made available with the addition of a modifier. It argues that an optimal account of such phenomena should derive these effects from the interaction of the meanings of individual expressions in the sentence rather than by positing a formal licensing relation between the modifier and the noun it modifies. The general approach is explicated by considering the distribution of bare plurals in Italian and the availability of generic readings for plural definites in English. The overall goal is a modest one. It seeks to draw attention to properties of modification that go beyond set intersection by focusing on phenomena that have not so far been closely scrutinized in the literature. It uses the particular results to draw some conclusions about questions of general interest.Licensing by modification Abstract: This paper looks at the phenomena of licensing by modification, whereby an unacceptable noun phrase is redeemed by the presence of a modifier or an unavailable reading for a noun phrase is made available with the addition of a modifier. It argues that an optimal account of such phenomena should derive these effects from the interaction of the meanings of individual expressions in the sentence rather than by positing a formal licensing relation between the modifier and the noun it modifies. The general approach is explicated by considering the distribution of bare plurals in Italian and the availability of generic readings for plural definites in English. The overall goal is a modest one. It seeks to draw attention to properties of modification that go beyond set intersection by focusing on phenomena that have not so far been closely scrutinized in the literature. It uses the particular results to draw some conclusions about questions of general

  8. Permit application modifications

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1995-11-01

    This document contains the Permit Application Modifications for the Y-12 Industrial Landfill V site on the Oak Ridge Reservation. These modifications include the assessment of stability of the proposed Landfill V under static and loading conditions. Analyses performed include the general slope stability, veneer stability of the bottom liner and cover system, and a liquefaction potential assessment of the foundation soils.

  9. Discovery of 1-[4-(N-benzylamino)phenyl]-3-phenylurea derivatives as non-peptidic selective SUMO-sentrin specific protease (SENP)1 inhibitors.

    Science.gov (United States)

    Uno, Masaharu; Koma, Yosuke; Ban, Hyun Seung; Nakamura, Hiroyuki

    2012-08-15

    We developed 1-[4-(N-benzylamino)phenyl]-3-phenylurea derivative 4 (GN6958) as a non-peptidic selective SUMO-sentrin specific protease (SENP)1 protease inhibitor based on the hypoxia inducible factor (HIF)-1α inhibitor 1 (GN6767). The direct interaction of compound 1 with SENP1 protein in cells was observed by the pull-down experiments using the biotin-tagged compound 2 coated on the streptavidin affinity column. Among the various 1-[4-(N-benzylamino)phenyl]-3-phenylurea derivatives tested, compounds 3 and 4 suppressed HIF-1α accumulation in a concentration-dependent manner without affecting the expression level of tubulin protein in HeLa cells. Both compounds inhibited SENP1 protease activity in a concentration-dependent manner, and compound 4 exhibited more potent inhibition than compound 3. Compound 4 exhibited selective inhibition against SENP1 protease activity without inhibiting other protease enzyme activities in vitro.

  10. Papel de SUMO en la regulación de PKR y la vía molecular PI3K/AKT

    OpenAIRE

    2015-01-01

    Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 17-07-2015 La SUMOilación es una modificación post-traduccional que consiste en que las proteínas SUMO se conjugan a sus proteínas diana. Esta modificación regula proteínas celulares involucradas en múltiples procesos como transcripción, replicación, reparación del ADN, apoptosis, senescencia, transporte nuclear y transducción de señales. De esta fo...

  11. Enzymatic modification of starch

    DEFF Research Database (Denmark)

    Jensen, Susanne Langgård

    substrate, and granular products were only obtained at low hydration of the starch. Hence, limiting hydration and gelatinization by using low-phosphate starch and high substrate oncentration was required for obtaining these products. Also high BE activity was a requirement and could partly compensate...... exclusively been conducted on gelatinized starch. This study provides a new concept for transferase-based modification of starches in granular state.......In the food industry approaches for using bioengineering are investigated as alternatives to conventional chemical and physical starch modification techniques in development of starches with specific properties. Enzyme-assisted post-harvest modification is an interesting approach to this, since...

  12. Benign Weather Modification

    Science.gov (United States)

    2007-11-02

    operational interest in modifying weather to support combat operations increased, ultimately leading to a multi-service effort called PROJECT POPEYE . The goal...This, coupled with the revelations concerning weather modification use in the Vietnam War (PROJECT POPEYE ), was a double blow to weather modification...AWS-TR-74-247, June 1984. Cobb, James T., Jr., et. al. Project Popeye : Final Report. China Lake, CA: Naval Weapons Center, 1967. Langmuir, Irving

  13. Antimicrobial Modifications of Polymers

    OpenAIRE

    Sedlarik, Vladimir

    2013-01-01

    This chapter is focused on antimicrobial modifications of polymer materials intended for medical devices production. Firstly, a brief introduction into the field of medical application of polymers is presented. Considering the fact that polymer medical devices are often connected with occurrence of nosocomial infections, the next part refers to this phenomenon and its causes. One of the possibilities of reducing of the infection occurrence is aimed at polymer modification. It is a key topic o...

  14. Modification as a propositional act

    DEFF Research Database (Denmark)

    Rijkhoff, Jan

    2014-01-01

    in Functional Discourse Grammar. Apart from Attitudinal Modification, there are at least five Subacts of Modification that are required for an adequate functional analysis of noun phrases and clauses: classifying modification, qualifying modification, quantifying modification, localizing modification......A complete functional theory of grammar should be able to specify the communicative function of each constituent in a sentence (or rather ‘Discourse Act’). Whereas predicating and referring have been given due attention in functional approaches to grammar, the modifying function of linguistic...

  15. Human Rights and Behavior Modification

    Science.gov (United States)

    Roos, Philip

    1974-01-01

    Criticisms of behavior modification, which charge that it violates ethical and legal principles, are discussed and reasons are presented to explain behavior modification's susceptibility to attack. (GW)

  16. Foot-and-mouth disease virus-like particles produced by a SUMO fusion protein system in Escherichia coli induce potent protective immune responses in guinea pigs, swine and cattle.

    Science.gov (United States)

    Guo, Hui-Chen; Sun, Shi-Qi; Jin, Ye; Yang, Shun-Li; Wei, Yan-Quan; Sun, De-Hui; Yin, Shuang-Hui; Ma, Jun-Wu; Liu, Zai-Xin; Guo, Jian-Hong; Luo, Jian-Xun; Yin, Hong; Liu, Xiang-Tao; Liu, Ding Xiang

    2013-07-04

    Foot-and-mouth disease virus (FMDV) causes a highly contagious infection in cloven-hoofed animals. The format of FMD virus-like particles (VLP) as a non-replicating particulate vaccine candidate is a promising alternative to conventional inactivated FMDV vaccines. In this study, we explored a prokaryotic system to express and assemble the FMD VLP and validated the potential of VLP as an FMDV vaccine candidate. VLP composed entirely of FMDV (Asia1/Jiangsu/China/2005) capsid proteins (VP0, VP1 and VP3) were simultaneously produced as SUMO fusion proteins by an improved SUMO fusion protein system in E. coli. Proteolytic removal of the SUMO moiety from the fusion proteins resulted in the assembly of VLP with size and shape resembling the authentic FMDV. Immunization of guinea pigs, swine and cattle with FMD VLP by intramuscular inoculation stimulated the FMDV-specific antibody response, neutralizing antibody response, T-cell proliferation response and secretion of cytokine IFN-γ. In addition, immunization with one dose of the VLP resulted in complete protection of these animals from homologous FMDV challenge. The 50% protection dose (PD50) of FMD VLP in cattle is up to 6.34. These results suggest that FMD VLP expressed in E. coli are an effective vaccine in guinea pigs, swine and cattle and support further development of these VLP as a vaccine candidate for protection against FMDV.

  17. Structural dynamic modification

    Indian Academy of Sciences (India)

    A Sestieri

    2000-06-01

    Vibration and acoustic requirements are becoming increasingly important in the design of mechanical structures, but they are not usually of primary concern in the design process. So the need to vary the structural behaviour to solve noise and vibration problems often occurs at the prototype stage, giving rise to the so-called structural modification problem. In this paper, the direct problem of determing the new response of a system, after some modifications are introduced into the sestem, is analysed using two different databases: the modal database and the frequency response function database. The limitaions of the modal database are discussed. Structural modifications that can be accounted for are lumped masses, springs, dampers and dynamic absorbers.

  18. C2株蓝氏贾第鞭毛虫SUMO基因的克隆、原核表达和生物信息学分析%Cloning, prokaryotic expression and bioinformatics analysis of Giardia lamblia C2 strain SUMO gene

    Institute of Scientific and Technical Information of China (English)

    王洋; 王沂; LI Wei-wei; 李冀; 李思瑾; 楚晗; 田喜凤

    2013-01-01

    Small ubiquitin-like modifier (or SUMO) proteins are a family of small proteins that are covalently attached to and detached from other proteins in cells to modify their function.In order to express Giardia lamblia (C2 strain) SUMO prorein in E.coli,the full-length open reading frame of SUMO was amplified by PCR from Giardia lamblia genome DNA.The PCR product about 320 bp in length was cloned into prokaryotic expression vector pET 28a(+) with restriction enzymes Nco I and Xho I.Sequencing result showed the sequence of SUMO in C2 strain was identical with that in WB strain.Bioinformatics analysis showed that SUMO protein displayed an ubiquitin-fold structure in three dimensional space.Phylogenetic analysis showed a distant relationship of SUMO protein between Giardia lamblia and other eukaryotes.The recombinant vector pET-28a(+)-SUMO was transformed into E.coli Rosetta(DE3),then the recombinant SUMO protein was expressed by IPTG induction.SDS-PAGE and western blot using anti-His Tag antibody showed that the expression product of SUMO was a fusion protein about 12.7 kD.The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SUMO protein provide basis for antibody preparation and functional study of SUMO related protein.%目的 克隆、原核表达C2株蓝氏贾第鞭毛虫(Giardia lamblia,简称贾第虫)的小泛素相关修饰物(small ubiquit in-related modifier,SUMO)蛋白,并对其序列进行生物信息学分析.方法 提取C2株贾第虫基因组DNA,以基因组DNA为模板PCR获得SUMO基因片段,双酶切连入原核表达载体pET28a(+),经测序验证并进行生物信息学分析,将重组质粒pET 28a(+)-SUMO转化大肠杆菌Rosetta(DE3).IPTG诱导后收集菌体,裂解后进行SDS-PAGE及Western blot检测.结果 成功构建了C2株贾第虫SUMO基因原核表达载体pET 28a(+)-SUMO,经IPTG诱导后,在大肠杆菌中高效表达,SDSPAGE及Western blot分析显示,在相对分子量约12.7 kD的位置出现目的蛋

  19. Organic modification of carbon nanotubes

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The organic modification of carbon nanotubes is a novel research field being developed recently. In this article, the history and newest progress of organic modification of carbon nanotubes are reviewed from two aspects:organic covalent modification and organic noncovalent modification of carbon nanotubes. The preparation and properties of organic modified carbon nanotubes are discussed in detail. In addition, the prospective development of organic modification of carbon nanotubes is suggested.

  20. Biblical behavior modification.

    Science.gov (United States)

    Lasure, L C; Mikulas, W L

    1996-07-01

    Although we may have formalized and systematized the field of behavior modification in the last few decades, people around the world have been using behavioral change strategies throughout history. Premack's (1965) theory of reinforcement is often called "Grandma's rule" because grandmothers have long been using it (e.g. You must finish your vegetables before you may go out and play). Franks (1969, p. 4), in one of the first behavioral texts, gave historical examples from China, Turkey, France, and Italy. Knapp and Shodahl (1974) showed how Benjamin Franklin used behavior modification. And de Silva (1984, 1985) gave examples of behavior modification by the Buddha and other early Buddhists. Conspicuously absent from our literature are examples from the Judeo-Christian tradition. In this paper, we provide a number of behavior modification examples from the Bible (New International Version). Footnotes provide references for many more examples. In the discussion, we explore implications for education and therapy. Examples are grouped by the following categories: operant conditioning, respondent conditioning, modeling, and cognitive interventions. However, the Biblical examples, like contemporary case studies, do not always fall neatly into discrete categories. They often are a combination, particularly operant and respondent conditioning interweaving.

  1. Context modification in action

    NARCIS (Netherlands)

    Visser, A.

    2008-01-01

    In this paper we develop the positive fragment of Context Modification Logic. This logical system is a variant of Dynamic Predicate Logic that employs multiple-access variables and that treats argument places in the same way as Latin. The positive fragment is purely incremental.

  2. A role for the Cajal-body-associated SUMO isopeptidase USPL1 in snRNA transcription mediated by RNA polymerase II.

    Science.gov (United States)

    Hutten, Saskia; Chachami, Georgia; Winter, Ulrike; Melchior, Frauke; Lamond, Angus I

    2014-03-01

    Cajal bodies are nuclear structures that are involved in biogenesis of snRNPs and snoRNPs, maintenance of telomeres and processing of histone mRNA. Recently, the SUMO isopeptidase USPL1 was identified as a component of Cajal bodies that is essential for cellular growth and Cajal body integrity. However, a cellular function for USPL1 is so far unknown. Here, we use RNAi-mediated knockdown in human cells in combination with biochemical and fluorescence microscopy approaches to investigate the function of USPL1 and its link to Cajal bodies. We demonstrate that levels of snRNAs transcribed by RNA polymerase (RNAP) II are reduced upon knockdown of USPL1 and that downstream processes such as snRNP assembly and pre-mRNA splicing are compromised. Importantly, we find that USPL1 associates directly with U snRNA loci and that it interacts and colocalises with components of the Little Elongation Complex, which is involved in RNAPII-mediated snRNA transcription. Thus, our data indicate that USPL1 plays a key role in RNAPII-mediated snRNA transcription.

  3. Detection of O-propargyl-puromycin with SUMO and ubiquitin by click chemistry at PML-nuclear bodies during abortive proteasome activities.

    Science.gov (United States)

    Uozumi, Naoki; Matsumoto, Hotaru; Saitoh, Hisato

    2016-05-27

    The amino-nucleoside antibiotic, puromycin, acts by covalently linking to elongating polypeptide chains on ribosomes to generate prematurely terminated immature polypeptides. The trafficking of puromycin-conjugated (puromycylated) immature polypeptides within cell has, however, remained elusive. In this study, using O-propargyl-puromycin (OP-Puro), the distribution of puromycylated polypeptides was assessed in HeLa cells by click chemistry. Under standard culture conditions, OP-Puro signals were detected in the cytoplasm and nucleus with the highest concentrations in the nucleolus. Intriguingly, when proteasome activities were aborted using MG132, OP-Puro signals began to accumulate at promyelocytic leukemia nuclear bodies (PML-NBs) in addition to the nucleolus. We also found promiscuous association of OP-Puro signals with SUMO-2/3 and ubiquitin at PML-NBs, but not at the nucleolus, during abortive proteasome activities. This study reveals a previously unknown distribution of OP-Puro that argues for a nuclear function in regulating immature protein homeostasis.

  4. Posttranslational protein modification in Archaea.

    Science.gov (United States)

    Eichler, Jerry; Adams, Michael W W

    2005-09-01

    One of the first hurdles to be negotiated in the postgenomic era involves the description of the entire protein content of the cell, the proteome. Such efforts are presently complicated by the various posttranslational modifications that proteins can experience, including glycosylation, lipid attachment, phosphorylation, methylation, disulfide bond formation, and proteolytic cleavage. Whereas these and other posttranslational protein modifications have been well characterized in Eucarya and Bacteria, posttranslational modification in Archaea has received far less attention. Although archaeal proteins can undergo posttranslational modifications reminiscent of what their eucaryal and bacterial counterparts experience, examination of archaeal posttranslational modification often reveals aspects not previously observed in the other two domains of life. In some cases, posttranslational modification allows a protein to survive the extreme conditions often encountered by Archaea. The various posttranslational modifications experienced by archaeal proteins, the molecular steps leading to these modifications, and the role played by posttranslational modification in Archaea form the focus of this review.

  5. Pectin modifications: a review.

    Science.gov (United States)

    Chen, Jun; Liu, Wei; Liu, Cheng-Mei; Li, Ti; Liang, Rui-Hong; Luo, Shun-Jing

    2015-01-01

    In recent years, the interest in studying modification of pectin has increased. A number of hydroxyl and carboxyl groups distributed along the backbone as well as a certain amount of neutral sugars presented as side chains make pectin capable of preparing a broad spectrum of derivatives. By forming pectin derivatives, their properties may be modified and some other new functional properties may be created. This article attempts to review the information about various methods used for pectin modification, including substitution (alkylation, amidation, quaternization, thiolation, sulfation, oxidation, etc.), chain elongation (cross-linking and grafting) and depolymerization (chemical, physical, and enzymatic degradation). Characteristics and applications of some pectin derivatives are also presented. In addition, the safety and regulatory status of pectin and its derivatives were reviewed.

  6. Geometrically Consistent Mesh Modification

    KAUST Repository

    Bonito, A.

    2010-01-01

    A new paradigm of adaptivity is to execute refinement, coarsening, and smoothing of meshes on manifolds with incomplete information about their geometry and yet preserve position and curvature accuracy. We refer to this collectively as geometrically consistent (GC) mesh modification. We discuss the concept of discrete GC, show the failure of naive approaches, and propose and analyze a simple algorithm that is GC and accuracy preserving. © 2010 Society for Industrial and Applied Mathematics.

  7. Genetic modification and genetic determinism.

    Science.gov (United States)

    Resnik, David B; Vorhaus, Daniel B

    2006-06-26

    In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound. Serious discussion of the morality of genetic modification, and the development of sound science policy, should be driven by arguments that address the actual consequences of genetic modification for individuals and society, not by ones propped up by false or misleading biological assumptions.

  8. Evaluation of a SUMO E2 conjugating enzyme involved in resistance to Clavibacter michiganensis subsp. michiganensis in Solanum peruvianum, through a tomato mottle virus VIGS assay

    Directory of Open Access Journals (Sweden)

    Mayra Janeth Esparza-Araiza

    2015-12-01

    Full Text Available Clavibacter michiganensis subsp. michiganensis (Cmm causes bacterial wilt and canker of tomato. Currently, no Solanum lycopersicum resistant varieties are commercially available, but some degree of Cmm resistance has been identified in Solanum peruvianum. Previous research showed up-regulation of a SUMO E2 conjugating enzyme (SCEI transcript in resistant S. peruvianum compared to susceptible S. lycopersicum following infection by Cmm. In order to test the role of SCEI in resistance to Cmm, a fragment of the gene from S. peruvianum was cloned into a novel virus-induced gene-silencing (VIGS vector based on the geminivirus Tomato Mottle Virus (ToMoV. Using biolistic inoculation, the ToMoV-based VIGS vector was shown to be effective in S. peruvianum by silencing the magnesium chelatase gene, which resulted in leaf bleaching. The ToMoV_SCEI construct resulted in approx. 61% silencing of SCEI in leaves of S. peruvianum as determined by quantitative RT-PCR. VIGS of SCEI in S. peruvianum resulted in unilateral wilting (15 dpi and subsequent death (20 dpi of the entire plant after Cmm inoculation, whereas empty vector-treated plants only showed wilting in the Cmm-inoculated leaf. SCEI-silenced plants also showed higher Cmm colonization with an average of 4.5 times more damaged tissue compared to the empty vector control plants. SCEI appears to play an important role in the innate immunity of S. peruvianum against Cmm, perhaps through the regulation of WRKY transcription factors, which may lead to expression of proteins involved in salicylic acid-dependent defense responses.

  9. Evaluation of a SUMO E2 Conjugating Enzyme Involved in Resistance to Clavibacter michiganensis Subsp. michiganensis in Solanum peruvianum, Through a Tomato Mottle Virus VIGS Assay.

    Science.gov (United States)

    Esparza-Araiza, Mayra J; Bañuelos-Hernández, Bernardo; Argüello-Astorga, Gerardo R; Lara-Ávila, José P; Goodwin, Paul H; Isordia-Jasso, María I; Castillo-Collazo, Rosalba; Rougon-Cardoso, Alejandra; Alpuche-Solís, Ángel G

    2015-01-01

    Clavibacter michiganensis subsp. michiganensis (Cmm) causes bacterial wilt and canker of tomato. Currently, no Solanum lycopersicum resistant varieties are commercially available, but some degree of Cmm resistance has been identified in Solanum peruvianum. Previous research showed up-regulation of a SUMO E2 conjugating enzyme (SCEI) transcript in S. peruvianum compared to S. lycopersicum following infection with Cmm. In order to test the role of SCEI in resistance to Cmm, a fragment of SCEI from S. peruvianum was cloned into a novel virus-induced gene-silencing (VIGS) vector based on the geminivirus, Tomato Mottle Virus (ToMoV). Using biolistic inoculation, the ToMoV-based VIGS vector was shown to be effective in S. peruvianum by silencing the magnesium chelatase gene, resulting in leaf bleaching. VIGS with the ToMoV_SCEI construct resulted in ~61% silencing of SCEI in leaves of S. peruvianum as determined by quantitative RT-PCR. The SCEI-silenced plants showed unilateral wilting (15 dpi) and subsequent death (20 dpi) of the entire plant after Cmm inoculation, whereas the empty vector-treated plants only showed wilting in the Cmm-inoculated leaf. The SCEI-silenced plants showed higher Cmm colonization and an average of 4.5 times more damaged tissue compared to the empty vector control plants. SCEI appears to play an important role in the innate immunity of S. peruvianum against Cmm, perhaps through the regulation of transcription factors, leading to expression of proteins involved in salicylic acid-dependent defense responses.

  10. Development and validation of a method for profiling post-translational modification activities using protein microarrays.

    Directory of Open Access Journals (Sweden)

    Sonia V Del Rincón

    Full Text Available BACKGROUND: Post-translational modifications (PTMs impact on the stability, cellular location, and function of a protein thereby achieving a greater functional diversity of the proteome. To fully appreciate how PTMs modulate signaling networks, proteome-wide studies are necessary. However, the evaluation of PTMs on a proteome-wide scale has proven to be technically difficult. To facilitate these analyses we have developed a protein microarray-based assay that is capable of profiling PTM activities in complex biological mixtures such as whole-cell extracts and pathological specimens. METHODOLOGY/PRINCIPAL FINDINGS: In our assay, protein microarrays serve as a substrate platform for in vitro enzymatic reactions in which a recombinant ligase, or extracts prepared from whole cells or a pathological specimen is overlaid. The reactions include labeled modifiers (e.g., ubiquitin, SUMO1, or NEDD8, ATP regenerating system, and other required components (depending on the assay that support the conjugation of the modifier. In this report, we apply this methodology to profile three molecularly complex PTMs (ubiquitylation, SUMOylation, and NEDDylation using purified ligase enzymes and extracts prepared from cultured cell lines and pathological specimens. We further validate this approach by confirming the in vivo modification of several novel PTM substrates identified by our assay. CONCLUSIONS/SIGNIFICANCE: This methodology offers several advantages over currently used PTM detection methods including ease of use, rapidity, scale, and sample source diversity. Furthermore, by allowing for the intrinsic enzymatic activities of cell populations or pathological states to be directly compared, this methodology could have widespread applications for the study of PTMs in human diseases and has the potential to be directly applied to most, if not all, basic PTM research.

  11. Protein interaction analysis of senataxin and the ALS4 L389S mutant yields insights into senataxin post-translational modification and uncovers mutant-specific binding with a brain cytoplasmic RNA-encoded peptide.

    Directory of Open Access Journals (Sweden)

    Craig L Bennett

    Full Text Available Senataxin is a large 303 kDa protein linked to neuron survival, as recessive mutations cause Ataxia with Oculomotor Apraxia type 2 (AOA2, and dominant mutations cause amyotrophic lateral sclerosis type 4 (ALS4. Senataxin contains an amino-terminal protein-interaction domain and a carboxy-terminal DNA/RNA helicase domain. In this study, we focused upon the common ALS4 mutation, L389S, by performing yeast two-hybrid screens of a human brain expression library with control senataxin or L389S senataxin as bait. Interacting clones identified from the two screens were collated, and redundant hits and false positives subtracted to yield a set of 13 protein interactors. Among these hits, we discovered a highly specific and reproducible interaction of L389S senataxin with a peptide encoded by the antisense sequence of a brain-specific non-coding RNA, known as BCYRN1. We further found that L389S senataxin interacts with other proteins containing regions of conserved homology with the BCYRN1 reverse complement-encoded peptide, suggesting that such aberrant protein interactions may contribute to L389S ALS4 disease pathogenesis. As the yeast two-hybrid screen also demonstrated senataxin self-association, we confirmed senataxin dimerization via its amino-terminal binding domain and determined that the L389S mutation does not abrogate senataxin self-association. Finally, based upon detection of interactions between senataxin and ubiquitin-SUMO pathway modification enzymes, we examined senataxin for the presence of ubiquitin and SUMO monomers, and observed this post-translational modification. Our senataxin protein interaction study reveals a number of features of senataxin biology that shed light on senataxin normal function and likely on senataxin molecular pathology in ALS4.

  12. Enzymatic Modification of Sphingomyelin

    DEFF Research Database (Denmark)

    Due to its major role in maintaining the water-retaining properties of the epidermis, ceramide is of great commercial potential in cosmetic and pharmaceuticals such as hair and skin care products. Currently, chemical synthesis of ceramide is a costly process, and developments of alternative cost......-efficient, high yield production methods are of great interest. In the present study, the potential of producing ceramide through the enzymatic hydrolysis of sphingomyelin have been studied. sphingomyelin is a ubiquitous membrane-lipid and rich in dairy products or by-products. It has been verified...... that sphingomyelin modification gives a feasible approach to the potential production of ceramide. The reaction system has been improved through system evaluation and the optimization of several important factors, and phospholipase C from Clostridium perfringens shows higher activity towards the hydrolysis reaction...

  13. End Group Modification

    DEFF Research Database (Denmark)

    Jahnsen, Rasmus O; Sandberg-Schaal, Anne; Frimodt-Møller, Niels

    2015-01-01

    Increased incidence of infections with multidrug-resistant bacterial strains warrants an intensive search for novel potential antimicrobial agents. Here, an antimicrobial peptide analogue with a cationic/hydrophobic alternating design displaying only moderate activity against Gram-positive pathog......Increased incidence of infections with multidrug-resistant bacterial strains warrants an intensive search for novel potential antimicrobial agents. Here, an antimicrobial peptide analogue with a cationic/hydrophobic alternating design displaying only moderate activity against Gram......-positive pathogens was optimized. Generally, introduction of hydrophobic moieties at the N-terminus resulted in analogues with remarkably increased activity against multidrug-resistant Staphylococcus aureus and Enterococcus faecium. Interestingly, the potency against Escherichia coli strains was unaffected, whereas...... modification with hydrophobic moieties led to increased activity towards the Gram-negative Acinetobacter baumannii. Despite increased cytotoxicity against murine fibroblasts and human umbilical vein endothelial cells, the optimized peptide analogues exhibited significantly improved cell selectivity. Overall...

  14. Surface modification technology

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Wonbaek; Yu, Hyosin; Chung, Inwha; Rhee, Kang In; Choi, Good Sun; Lee, Chulkyung; Youn, In Ju; Chung, Jinki; Suh, Chang Youl; Yang, Dong Hyo [Korea Institute of Geology Mining and Materials, Taejon (Korea, Republic of)

    1997-12-01

    Domestic production of rare metals has not been achieved due to the low metal content in their ores in the nation. For these reasons, a strategy for the value addition of rare metal sponges by processes like vacuum melting should be seek to meet the growing domestic demand for titanium metals and alloys. However, current domestic market appears not to be sufficient enough for the large scale investment for the expensive vacuum-melting equipment. Besides, related ingot-processing technologies like rolling, extrusion, and forging of titanium should be prepared in advance. In the mean time, the attempt to recycle expensive titanium scraps produced in our nation would be worthwhile in view of the reduction of import from foreign countries and of saving valuable secondary resources. The objectives for this research is to develop a multipurpose inductively-coupled plasma enhanced-surface modification (ICPESM) process to upgrade powder products. Stable plasma was obtained by the impedance harmonization between plasma generator and matching networks maintaining the reflected power at near zero. The chamber vacuum went down to 10{sup -3} torr offering no difficulties to maintain 1.0 torr at which the present experiments were conducted. However, the fluidization in the chamber was unstable when operated in vacuum. The gas distributor and chamber design may need modifications. Argon plasma treatment on the titanium powders changed the surface morphology slightly even though the effect was not significant due possibly to the short treatment duration of 60 minutes. Oxygen plasma oxidized the surface of titanium powders to TiO{sub 2} as confirmed by XRD. The carbon black powders were clustered during oxidation treatment by the fluidization or surface activation by the high power of low temperature plasma. (author). 4 tabs., 15 figs.

  15. Overexpression of the Rice SUMO E3 Ligase Gene OsSIZ1 in Cotton Enhances Drought and Heat Tolerance, and Substantially Improves Fiber Yields in the Field under Reduced Irrigation and Rainfed Conditions

    Science.gov (United States)

    Mishra, Neelam; Sun, Li; Zhu, Xunlu; Smith, Jennifer; Prakash Srivastava, Anurag; Yang, Xiaojie; Pehlivan, Necla; Esmaeili, Nardana; Luo, Hong; Shen, Guoxin; Jones, Don; Auld, Dick; Burke, John

    2017-01-01

    The Arabidopsis SUMO E3 ligase gene AtSIZ1 plays important roles in plant response to abiotic stresses as loss of function in AtSIZ1 leads to increased sensitivity to drought, heat and salt stresses. Overexpression of the AtSIZ1 rice homolog, OsSIZ1, leads to increased heat and drought tolerance in bentgrass, suggesting that the function of the E3 ligase SIZ1 is highly conserved in plants and it plays a critical role in abiotic stress responses. To test the possibility that the SUMO E3 ligase could be used to engineer drought- and heat-tolerant crops, the rice gene OsSIZ1 was overexpressed in cotton. We report here that overexpression of OsSIZ1 in cotton results in higher net photosynthesis and better growth than wild-type cotton under drought and thermal stresses in growth chamber and greenhouse conditions. Additionally, this tolerance to abiotic stresses was correlated with higher fiber yield in both controlled-environment and field trials carried out under reduced irrigation and rainfed conditions. These results suggest that OsSIZ1 is a viable candidate gene to improve crop yields under water-limited and rainfed agricultural production systems. PMID:28340002

  16. Genetic modification and genetic determinism

    OpenAIRE

    Vorhaus Daniel B; Resnik David B

    2006-01-01

    Abstract In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound....

  17. SUMO1基因单核苷酸多态性分析及rs7599810多态性与非综合征型唇/腭裂的关联研究%Analysis of the HapMap data on SNPs in SUMO1 and association study of rs7599810 in trios with non-syndromic cleft lip with or without cleft palate

    Institute of Scientific and Technical Information of China (English)

    岳青; 王红; 张博; 赵凯平

    2014-01-01

    目的:对HapMap数据库四个不同人群SUMO1基因的单核苷酸多态性(single nucleotide polymorphisms,SNPs)进行分析和比较,并探讨山东汉族人群rs7599810位点多态性与非综合征型唇/腭裂(non-syndromic cleft lip with or without cleft palate,NSCL/P)的关联.方法:首先用Haploview软件确定HapMap四个人群,分别为:中国北京汉族(Han Chinese in Beijing,China,CHB)、日本东京(Japanese in Tokyo,Japan,JPT)、西北欧后裔(Utah residents with Northern and Western European ancestry from the CEPH collection,CEU)和尼日利亚约鲁巴(Yoruban in Ibadan,Nigeria,YRI),四个人群中SUMO1基因SNPs中最小等位基因频率(minor allele frequency,MAF)>0.01且符合Hardy-Weinberg平衡的SNPs为合格SNPs,用MAF全距相对比指标衡量人群间最小等位基因相同的共有合格SNPs的MAFs差异;然后对四个人群共有合格SNPs进行单体域和单体型的分析及比较;最后对本研究所测该基因中的rs7599810在山东地区183个NSCL/P核心家系中进行传递不平衡检验(transmission/disequilibrium test,TDT).结果:HapMap提供的24个SNPs中,纯合SNPs在CHB和JPT均为9个,在CEU为8个,在YRI为6个,四个人群共有合格SNPs为12个.这些SNPs在四个人群均形成了一个单体域,其中CHB、JPT、CEU人群的单体域包括全部12个SNPs,YRI人群的单体域包括第3~11个SNPs.CHB、JPT、CEU和YRI人群的单体域分别确定了3、3、5、6种单体型,且均以前两种单体型为主,频率为0.634~ 0.922.TDT结果显示SUMO1基因rs7599810多态性与NSCL/P未见统计学关联(x2 =0.485,P=0.486,OR=0.898,95% CI:0.663 ~1.021).结论:CHB、JPT和CEU人群SUMO1基因SNPs在等位基因组成、MAF、单体域及单体型构成等方面以共性特征为主,与之相比,YRI人群SNPs表现出较大的独特性.山东汉族人群中SUMO1基因rs7599810多态性与NSCL/P未见统计学关联.

  18. Glucocorticoid-induced tethered transrepression requires SUMOylation of GR and formation of a SUMO-SMRT/NCoR1-HDAC3 repressing complex

    Science.gov (United States)

    Hua, Guoqiang; Ganti, Krishna Priya; Chambon, Pierre

    2016-01-01

    Upon binding of a glucocorticoid (GC), the GC receptor (GR) can exert one of three transcriptional regulatory functions. We recently reported that SUMOylation of the GR at position K293 in humans (K310 in mice) within the N-terminal domain is indispensable for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression. We now demonstrate that the integrity of this GR SUMOylation site is mandatory for the formation of a GR-small ubiquitin-related modifiers (SUMOs)-SMRT/NCoR1-HDAC3 repressing complex, which is indispensable for NF-κB/AP1-mediated GC-induced tethered indirect transrepression in vitro. Using GR K310R mutant mice or mice containing the N-terminal truncated GR isoform GRα-D3 lacking the K310 SUMOylation site, revealed a more severe skin inflammation than in WT mice. Importantly, cotreatment with dexamethasone (Dex) could not efficiently suppress a 12-O-tetradecanoylphorbol-13-acetate (TPA)–induced skin inflammation in these mutant mice, whereas it was clearly decreased in WT mice. In addition, in mice selectively ablated in skin keratinocytes for either nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors or histone deacetylase 3 (HDAC3), Dex-induced tethered transrepression and the formation of a repressing complex on DNA-bound NF-κB/AP1 were impaired. We previously suggested that GR ligands that would lack both (+)GRE-mediated transactivation and IR nGRE-mediated direct transrepression activities of GCs may preferentially exert the therapeutically beneficial GC antiinflammatory properties. Interestingly, we now identified a nonsteroidal antiinflammatory selective GR agonist (SEGRA) that selectively lacks both Dex-induced (+)GRE-mediated transactivation and IR nGRE-mediated direct transrepression functions, while still exerting a tethered indirect transrepression activity and could therefore be clinically lesser

  19. GR SUMOylation and formation of an SUMO-SMRT/NCoR1-HDAC3 repressing complex is mandatory for GC-induced IR nGRE-mediated transrepression

    Science.gov (United States)

    Hua, Guoqiang; Paulen, Laetitia; Chambon, Pierre

    2016-01-01

    Unique among the nuclear receptor superfamily, the glucocorticoid (GC) receptor (GR) can exert three distinct transcriptional regulatory functions on binding of a single natural (cortisol in human and corticosterone in mice) and synthetic [e.g., dexamethasone (Dex)] hormone. The molecular mechanisms underlying GC-induced positive GC response element [(+)GRE]-mediated activation of transcription are partially understood. In contrast, these mechanisms remain elusive for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression and for tethered indirect transrepression that is mediated by DNA-bound NF-κB/activator protein 1 (AP1)/STAT3 activators and instrumental in GC-induced anti-inflammatory activity. We demonstrate here that SUMOylation of lysine K293 (mouse K310) located within an evolutionary conserved sequence in the human GR N-terminal domain allows the formation of a GR-small ubiquitin-related modifiers (SUMOs)-NCoR1/SMRT-HDAC3 repressing complex mandatory for GC-induced IR nGRE-mediated direct repression in vitro, but does not affect transactivation. Importantly, these results were validated in vivo: in K310R mutant mice and in mice ablated selectively for nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors in skin keratinocytes, Dex-induced direct repression and the formation of repressing complexes on IR nGREs were impaired, whereas transactivation was unaffected. In mice selectively ablated for histone deacetylase 3 (HDAC3) in skin keratinocytes, GC-induced direct repression, but not bindings of GR and of corepressors NCoR1/SMRT, was abolished, indicating that HDAC3 is instrumental in IR nGRE-mediated repression. Moreover, we demonstrate that the binding of HDAC3 to IR nGREs in vivo is mediated through interaction with SMRT/NCoR1. We also show that the GR ligand binding domain (LBD) is not required for SMRT

  20. GR SUMOylation and formation of an SUMO-SMRT/NCoR1-HDAC3 repressing complex is mandatory for GC-induced IR nGRE-mediated transrepression.

    Science.gov (United States)

    Hua, Guoqiang; Paulen, Laetitia; Chambon, Pierre

    2016-02-02

    Unique among the nuclear receptor superfamily, the glucocorticoid (GC) receptor (GR) can exert three distinct transcriptional regulatory functions on binding of a single natural (cortisol in human and corticosterone in mice) and synthetic [e.g., dexamethasone (Dex)] hormone. The molecular mechanisms underlying GC-induced positive GC response element [(+)GRE]-mediated activation of transcription are partially understood. In contrast, these mechanisms remain elusive for GC-induced evolutionary conserved inverted repeated negative GC response element (IR nGRE)-mediated direct transrepression and for tethered indirect transrepression that is mediated by DNA-bound NF-κB/activator protein 1 (AP1)/STAT3 activators and instrumental in GC-induced anti-inflammatory activity. We demonstrate here that SUMOylation of lysine K293 (mouse K310) located within an evolutionary conserved sequence in the human GR N-terminal domain allows the formation of a GR-small ubiquitin-related modifiers (SUMOs)-NCoR1/SMRT-HDAC3 repressing complex mandatory for GC-induced IR nGRE-mediated direct repression in vitro, but does not affect transactivation. Importantly, these results were validated in vivo: in K310R mutant mice and in mice ablated selectively for nuclear receptor corepressor 1 (NCoR1)/silencing mediator for retinoid or thyroid-hormone receptors (SMRT) corepressors in skin keratinocytes, Dex-induced direct repression and the formation of repressing complexes on IR nGREs were impaired, whereas transactivation was unaffected. In mice selectively ablated for histone deacetylase 3 (HDAC3) in skin keratinocytes, GC-induced direct repression, but not bindings of GR and of corepressors NCoR1/SMRT, was abolished, indicating that HDAC3 is instrumental in IR nGRE-mediated repression. Moreover, we demonstrate that the binding of HDAC3 to IR nGREs in vivo is mediated through interaction with SMRT/NCoR1. We also show that the GR ligand binding domain (LBD) is not required for SMRT

  1. Surface modification of cellulose nanocrystals

    Science.gov (United States)

    Eyley, Samuel; Thielemans, Wim

    2014-06-01

    Chemical modification of cellulose nanocrystals is an increasingly popular topic in the literature. This review analyses the type of cellulose nanocrystal modification reactions that have been published in the literature thus far and looks at the steps that have been taken towards analysing the products of the nanocrystal modifications. The main categories of reactions carried out on cellulose nanocrystals are oxidations, esterifications, amidations, carbamations and etherifications. More recently nucleophilic substitutions have been used to introduce more complex functionality to cellulose nanocrystals. Multi-step modifications are also considered. This review emphasizes quantification of modification at the nanocrystal surface in terms of degree of substitution and the validity of conclusions drawn from different analysis techniques in this area. The mechanisms of the modification reactions are presented and considered with respect to the effect on the outcome of the reactions. While great strides have been made in the quality of analytical data published in the field of cellulose nanocrystal modification, there is still vast scope for improvement, both in data quality and the quality of analysis of data. Given the difficulty of surface analysis, cross-checking of results from different analysis techniques is fundamental for the development of reliable cellulose nanocrystal modification techniques.

  2. Surface Modification of Intraocular Lenses

    Directory of Open Access Journals (Sweden)

    Qi Huang

    2016-01-01

    Full Text Available Objective: This paper aimed to review the current literature on the surface modification of intraocular lenses (IOLs. Data Sources: All articles about surface modification of IOLs published up to 2015 were identified through a literature search on both PubMed and ScienceDirect. Study Selection: The articles on the surface modification of IOLs were included, but those on design modification and surface coating were excluded. Results: Technology of surface modification included plasma, ion beam, layer-by-layer self-assembly, ultraviolet radiation, and ozone. The main molecules introduced into IOLs surface were poly (ethylene glycol, polyhedral oligomeric silsesquioxane, 2-methacryloyloxyethyl phosphorylcholine, TiO 2 , heparin, F-heparin, titanium, titanium nitride, vinyl pyrrolidone, and inhibitors of cytokines. The surface modification either resulted in a more hydrophobic lens, a more hydrophilic lens, or a lens with a hydrophilic anterior and hydrophobic posterior surface. Advances in research regarding surface modification of IOLs had led to a better biocompatibility in both in vitro and animal experiments. Conclusion: The surface modification is an efficient, convenient, economic and promising method to improve the biocompatibility of IOLs.

  3. Genetic modification and genetic determinism

    Directory of Open Access Journals (Sweden)

    Vorhaus Daniel B

    2006-06-01

    Full Text Available Abstract In this article we examine four objections to the genetic modification of human beings: the freedom argument, the giftedness argument, the authenticity argument, and the uniqueness argument. We then demonstrate that each of these arguments against genetic modification assumes a strong version of genetic determinism. Since these strong deterministic assumptions are false, the arguments against genetic modification, which assume and depend upon these assumptions, are therefore unsound. Serious discussion of the morality of genetic modification, and the development of sound science policy, should be driven by arguments that address the actual consequences of genetic modification for individuals and society, not by ones propped up by false or misleading biological assumptions.

  4. Chemical Protein Modification through Cysteine.

    Science.gov (United States)

    Gunnoo, Smita B; Madder, Annemieke

    2016-04-01

    The modification of proteins with non-protein entities is important for a wealth of applications, and methods for chemically modifying proteins attract considerable attention. Generally, modification is desired at a single site to maintain homogeneity and to minimise loss of function. Though protein modification can be achieved by targeting some natural amino acid side chains, this often leads to ill-defined and randomly modified proteins. Amongst the natural amino acids, cysteine combines advantageous properties contributing to its suitability for site-selective modification, including a unique nucleophilicity, and a low natural abundance--both allowing chemo- and regioselectivity. Native cysteine residues can be targeted, or Cys can be introduced at a desired site in a protein by means of reliable genetic engineering techniques. This review on chemical protein modification through cysteine should appeal to those interested in modifying proteins for a range of applications.

  5. RNA modification in Cajal bodies.

    Science.gov (United States)

    Meier, U Thomas

    2016-10-24

    Aside from nucleoli, Cajal bodies (CBs) are the best-characterized organelles of mammalian cell nuclei. Like nucleoli, CBs concentrate ribonucleoproteins (RNPs), in particular, spliceosomal small nuclear RNPs (snRNPs) and small nucleolar RNPs (snoRNPs). In one of the best-defined functions of CBs, most of the snoRNPs are involved in site-specific modification of snRNAs. The two major modifications are pseudouridylation and 2'-O-methylation that are guided by the box H/ACA and C/D snoRNPs, respectively. This review details the modifications, their function, the mechanism of modification, and the machineries involved. We dissect the different classes of noncoding RNAs that meet in CBs, guides and substrates. Open questions and conundrums, often raised and appearing due to experimental limitations, are pointed out and discussed. The emphasis of the review is on mammalian CBs and their function in modification of noncoding RNAs.

  6. Non-fusion soluble expression of broad-spectrum antivirus protein in Escherichia coli by translational-coupling with SUMO%利用翻译偶联实现广谱抗病毒蛋白在大肠杆菌中的可溶性表达

    Institute of Scientific and Technical Information of China (English)

    邢伶越; 谢德健; 叶丙雨; 张章; 李平; 沈文龙; 师明磊; 张彦; 赵志虎

    2015-01-01

    目的:以具有广泛助溶活性的小泛素相关修饰蛋白( small ubiquitin-related modifier,SUMO)为辅助蛋白,利用翻译偶联,构建一种新型非融合可溶原核表达载体,实现广谱抗病毒蛋白RA在大肠杆菌中的非融合可溶表达。方法以PCR方法从酵母基因组中克隆SUMO蛋白基因smt3并进行定点突变,去除其中的EcoRⅠ酶切位点,获得同义突变体mSUMO。通过翻译偶联Linker序列的设计、合成,构建以mSUMO蛋白为辅助蛋白的翻译偶联表达载体pTIG-mSUMO。 PCR扩增获得带有相应酶切位点的广谱抗病毒蛋白 RA 的基因,将其克隆至 pTIG-mSUMO中,获得表达载体pTIG-mSUMO/RA,实现RA在大肠杆菌中的非融合可溶性表达。结果 IPTG诱导蛋白表达以后,SDS-PAGE和Western印迹检测证实,目的蛋白RA的表达量明显增加,且可溶比例约占50%。结论以优化后的SUMO为辅助蛋白,成功构建了一种非融合的可溶原核表达载体pTIG-mSUMO,实现了RA蛋白在大肠杆菌中的高效非融合可溶表达,既提高了表达水平,也提高了可溶性。 pTIG-mSUMO作为一种可溶的非融合表达载体,在实现外源蛋白的高效可溶表达方面,很可能具有一定的普遍性。%Objective To design and construct a new non-fusion soluble expression vector pTIG-mSUMO(small ubiq-uitin-related modifier) using the widely used solubility promoting protein SUMO and based on the translational coupling phenomenon in order to enable the non-fusion soluble expression of the broad-spectrum antiviral protein RA in Escherichia coli by pTIG-mSUMO.Methods The smt3 gene coding for SUMO protein was cloned from yeast genome DNA by PCR. After directed-site silent mutation to eliminate the EcoRⅠsite, the mutant mSUMO was inserted into pET-22b to obtain the translational coupling expression vector pTIG-mSUMO.The RA was subject to PCR amplification and cloned into the pTIG-mSUMO to

  7. Surface modification of bioceramics

    Science.gov (United States)

    Monkawa, Akira

    Hydroxyapatite [Ca10(PO4)6(OH)2, HAp] is a major inorganic component of bone and teeth tissues and has the excellent biocompatibility and high osteoconductivity. The interactions between HAp and protein or cell have been studied. The HAp related bioceramics such as bone substitute, coating substance of metal implants, inorganic-polymer composites, and cell culture. We described two methods; (1) surface modification of HAp using organosilane; (2) fabrication of HAp ultra-thin layer on gold surface for protein adsorption analyzed with QCM-D technique. The interfacial interaction between collagen and HAp in a nano-region was controlled by depositing the organosilane of n-octadecyltrimethoxysilane (ODS: -CH3) or aminopropyltriethoxysilane (APTS: -NH2) with a chemical vapor deposition method. The morphologies of collagen adsorbed on the surfaces of HAp and HAp deposited with APTS were similar, however that of the surface with ODS was apparently different, due to the hydrophobic interaction between the organic head group of -CH3 and residual groups of collagen. We present a method for coating gold quartz crystal microbalance with dissipation (QCM-D) sensor with ultra-thin layer of hydroxyapatite nanocrystals evenly covering and tightly bound to the surface. The hydroxyapatite sensor operated in liquid with high stability and sensitivity. The in-situ adsorption mechanism and conformational change of fibrinogen on gold, titanium and hydroxyapatite surfaces were investigated by QCM-D technique and Fourier-transform infrared spectroscopy. The study indicates that the hydroxyapatite sensor is applicable for qualitative and conformational analysis of protein adsorption.

  8. System Evolution by Metalevel Modification

    Directory of Open Access Journals (Sweden)

    Michal Vagač

    2009-05-01

    Full Text Available This paper describes system evolution managed by corresponding metasystem. The metasystem builds a metamodel of base system and allows its modification. The modification is propagated back to the base system. The application model presents the example of standard graphics user interface developed with Java Abstract Windowing Toolkit (AWT, which is a part of the Java Foundation Classes (JFC. The main aim is to confirm the possibility of application properties monitoring using aspect-oriented programming, their abstraction in ametamodel, possibility of their alternations by metamodel modifications and consequent change in the original application model.

  9. SUMO 特异性蛋白酶1在心肌组织缺血/再灌注损伤中的保护作用%Protective Effect of SUMO-Specific Protease 1 on Ischemia/Reperfusion Injury of Myocardial Tissue

    Institute of Scientific and Technical Information of China (English)

    顾剑民; 薛松

    2014-01-01

    目的:探讨SUMO特异性蛋白酶1( SENP1)在心肌组织缺血/再灌注损伤中的保护作用。方法选取14例行体外循环心脏手术前、后的右心房组织,建立缺血再灌注小鼠模型,通过实时定量PCR检测患者和大鼠心肌组织SENP1的mRNA变化。对H9 C2细胞株进行缺氧复氧处理后检测 LDH释放率,观察细胞死亡情况。结果实时定量PCR结果显示,14例患者缺血再灌注后心肌组织内SENP1 mRNA含量为(4.845±1.248),较灌注前明显升高(P<0.01)。单纯缺血处理的小鼠心肌SENP1 mRNA含量较正常组小鼠略高,且随着灌注时间的延长,SENP1 mRNA含量呈逐渐递增的趋势。特异性siRNA敲除大鼠心肌细胞H9C2细胞内SENP148 h后SENP1 mRNA水平降至对照组的30%以下,敲除SENP1的H9C2细胞在缺氧及复氧处理后LDH释放增加。结论 SENP1在心肌缺血再灌注中有保护作用,缺乏SENP1可能会加重心肌损伤。%Objective To investigate the protective effect of SUMO -specific protease 1 ( SUMO-SENP1 ) on ischemia/reperfusion injury of myocardial tissue .Methods Fourteen cases of right atrial tissue before and after cardiopulmonary bypass heart operation were selected , the mouse model of ischemia reperfusion was established , and real-time quantitative polymerase chain reaction ( PCR) was used to detect the changes of SENP1 mRNA in myocardial tissue of patients and rats.The LDH release rate was detected after hypoxia reoxygenation on H 9C2 cell line,and the cell death was observed .Results The results of real-time PCR showed that the SENP1 mRNA content in myocardial tissue of14 cases after reperfusion was(4.845 ±1.248),which was significantly higher than that before reperfusion (P<0.01).The SENP1 mRNA content in myocardium of ischemia treatment rats was slightly higher than that of normal rats , and increased with the increasing reperfusion time .The mRNA level of SENP1 in H9C2 cells of specific si

  10. Respiratory psychophysiology and behavior modification.

    Science.gov (United States)

    Ley, R

    2001-09-01

    This article was written as an introduction to a special issue of Behavior Modification dedicated to studies in the field of respiratory psychophysiology. Although the invited articles that constitute this special issue cover a fairly broad range of topics, priority was given to articles that focus on the role of respiration in panic disorder. Attention is directed to the fundamental role of breathing in applied psychophysiology and to the encouragement of research in the modification of breathing behavior. The connection between respiratory psychophysiology and behavior modification is explained by reference to (a) a recent article on Pavlovian and operant control of breathing behavior and (b) four published volumes of selected articles dedicated exclusively to the field of respiratory psychophysiology. The present special issue of Behavior Modification marks the fifth volume.

  11. HMG Modifications and Nuclear Function

    OpenAIRE

    Zhang, Qingchun; Wang, Yinsheng

    2010-01-01

    High mobility group (HMG) proteins assume important roles in regulating chromatin dynamics, transcriptional activities of genes and other cellular processes. Post-translational modifications of HMG proteins can alter their interactions with DNA and proteins, and consequently, affect their biological activities. Although the mechanisms through which these modifications are involved in regulating biological processes in different cellular contexts are not fully understood, new insights into the...

  12. Posttranslational Protein Modification in Archaea

    OpenAIRE

    Eichler, Jerry; Adams, Michael W. W.

    2005-01-01

    One of the first hurdles to be negotiated in the postgenomic era involves the description of the entire protein content of the cell, the proteome. Such efforts are presently complicated by the various posttranslational modifications that proteins can experience, including glycosylation, lipid attachment, phosphorylation, methylation, disulfide bond formation, and proteolytic cleavage. Whereas these and other posttranslational protein modifications have been well characterized in Eucarya and B...

  13. Modification Management for Construction Processes

    Institute of Scientific and Technical Information of China (English)

    Wolfgang Huhnt; Sven Richter; Felix Enge

    2008-01-01

    This paper presents an approach to handle different types of subsequent modifications in con-struction processes. It is focused on a formalized approach with the objective that effects of subsequent modifications are detected automatically. The approach presented in this paper makes use of an existing modeling technique specifically focused on construction processes. The modeling techniques result in a process description where all technological interdependencies between construction activities are consid-ered and where consistency between the sequence of activities and the history of components is guaran-teed. This modeling technique is expanded by an existing concept for handling versions and dependencies between objects. The integration of these different concepts makes it possible to track effects of relevant types of subsequent modifications in construction processes on a formalized basis. Effects of subsequent modifications are detected by algorithms, different versions can be compared by algorithms and different versions are stored so that subsequent modifications are documented. The innovation of the approach pre-sented is the completeness. Effects of different relevant types of subsequent modifications in construction processes are tracked on a formalized basis.

  14. Weather Modification: Finding Common Ground.

    Science.gov (United States)

    Garstang, Michael; Bruintjes, Roelof; Serafin, Robert; Orville, Harold; Boe, Bruce; Cotton, William; Warburton, Joseph

    2005-05-01

    Research and operational approaches to weather modification expressed in the National Research Council's 2003 report on “Critical Issues in Weather Modification Research” and in the Weather Modification Association's response to that report form the basis for this discussion. There is agreement that advances in the past few decades over a broad front of understanding physical processes and in technology have not been comprehensively applied to weather modification. Such advances need to be capitalized upon in the form of a concerted and sustained national effort to carry out basic and applied research in weather modification. The need for credible scientific evidence and the pressure for action should be resolved. Differences in the perception of current knowledge, the utility of numerical models, and the specific needs of research and operations in weather modification must be addressed. The increasing demand for water and the cost to society inflicted by severe weather require that the intellectual, technical, and administrative resources of the nation be combined to resolve whether and to what degree humans can influence the weather.The National Center for Atmospheric Research is sponsored by the National Science Foundation

  15. Laser modification of polyamide fabrics

    Science.gov (United States)

    Bahtiyari, M. İ.

    2011-02-01

    A new method for the modification of the properties of polyamide fabric, based on exposure to the output from a CO 2 laser, has been investigated. It was found that, after laser modification of polyamide fabric, the dyeability of fabric was increased significantly, while the bursting strength was decreased. The reasons for this drastic increase in dyeability of polyamide fabrics have been analyzed with the help of FTIR and iodine sorption methods, revealing a relationship with a decrease in the crystallinity of the polyamide. It was observed that, as the laser modification of the fabric was carried out with low intensity, the concentration of free amino groups, which are necessary during dyeing with acid and reactive dyes, increased.

  16. Functionalised Polymers by Surface Modification

    Institute of Scientific and Technical Information of China (English)

    Jon-Paul Griffiths; M. G. Moloney

    2005-01-01

    @@ 1Introduction Surface-active polymers are of substantial importance in many diverse aspects of modern technology, with applications ranging from solid phase chemical synthesis related to drug discovery and chemical catalysis to biocompatible/bioactive medical implants and prostheses, and to surface-modified fabrics. Whilst there are a number of existing physical (e. g. corona or plasma discharge, ion beam irradiation[1] ) and chemical (e. g.silanisation, oxidation, chlorination, acylation and quaternisation[2-4]) methods for the surface modification of polymers, the frequent requirement for significant infrastructure, harsh reaction conditions, and limitation to specific polymer types (e. g. polybutadiene[5] ), which must possess suitable chemical functionality capable of direct modification, led us to consider alternative chemical methods. Desirable was an alternative that would be amenable to a large range of polymers, permitting direct chemical modification under mild conditions and using inexpensive reagents.

  17. Surface modification for corrosion resistance

    Energy Technology Data Exchange (ETDEWEB)

    Natesan, K.

    1993-06-01

    The raw gas environments that arise from coal gasification have chemical compositions that are low in pO{sub 2} and moderate-to-high in pS{sub 2}. Metallic materials for service in such an environment undergo predominantly sulfidation attack at temperatures of 400 to 700{degree}C. Modification of alloy compositions in bulk can alter the scaling processes and lead to improvements in corrosion resistance, but the benefits can only be attained at temperatures much higher than the service temperatures of the components. Modification of surfaces of structural components by several of the coating techniques examined in this study showed substantial benefit in corrosion resistance when tested in simulated coal gasification environments. The paper presents several examples of surface modification and their corrosion performance.

  18. Modification on epoxy-based adhesive

    Institute of Scientific and Technical Information of China (English)

    ZhengXiaoxia; QianChunxiang

    2003-01-01

    This research adopted four methods to toughen epoxy adhesives. They were liquid hydroxyl group terminated polybutadiene (HTPB) rubber modification, silicon rubber modification, polyacrylate multiplicity elastomer particulates emulsion modification and chemical grafting modification. After modification, the shearing strength and the rapture elongation were tested. The interface and the chemical reaction between the modifiers and the epoxy were analyzed by scanning electron microscope (SEM) and infrared optical spectrum. The results show that the elastomer particulates modification and the chemical grafting modification can reach the better toughening effects.

  19. [Targeted genome modifications using TALEN].

    Science.gov (United States)

    Dupret, Barbara; Angrand, Pierre-Olivier

    2014-02-01

    Precise modifications of genomes have been one of the biggest goals in the fields of biotechnology and biomedical research. Recent discovery of TALE (transcription activator-like effectors) and the engineering of customized TALEN (transcription activator-like effector nucleases) allowed rapid genome editing in a variety of cell types and different model organisms. TALEN are molecular scissors used to induce a wide range of specific and efficient genomic modifications. TALEN promise to have profound impacts on biological and medical research over the coming years.

  20. Structural dynamic modifications via models

    Indian Academy of Sciences (India)

    T K Kundra

    2000-06-01

    Structural dynamic modification techniques attempt to reduce dynamic design time and can be implemented beginning with spatial models of structures, dynamic test data or updated models. The models assumed in this discussion are mathematical models, namely mass, stiffness, and damping matrices of the equations of motion of a structure. These models are identified/extracted from dynamic test data viz. frequency response functions (FRFs). Alternatively these models could have been obtained by adjusting or updating the finite element model of the structure in the light of the test data. The methods of structural modification for getting desired dynamic characteristics by using modifiers namely mass, beams and tuned absorbers are discussed.

  1. Plasma surface modification of polymers

    Science.gov (United States)

    Hirotsu, T.

    1980-01-01

    Thin plasma polymerization films are discussed from the viewpoint of simplicity in production stages. The application of selective, absorbent films and films used in selective permeability was tested. The types of surface modification of polymers discussed are: (1) plasma etching, (2) surface coating by plasma polymerized thin films, and (3) plasma activation surface graft polymerization.

  2. Electrostatic modification of novel materials

    NARCIS (Netherlands)

    Ahn, C.H.; Bhattacharya, A.; Di Ventra, M.; Eckstein, J.N.; Frisbie, C.D.; Gershenson, M.E.; Goldman, A.M.; Inoue, I.H.; Mannhart, J.; Millis, A.J.; Morpurgo, A.F.; Natelson, D.; Triscone, J.M.

    2006-01-01

    Application of the field-effect transistor principle to novel materials to achieve electrostatic doping is a relatively new research area. It may provide the opportunity to bring about modifications of the electronic and magnetic properties of materials through controlled and reversible changes of t

  3. Behavior Modification in Marriage Counseling

    Science.gov (United States)

    Baird, Emily; Redfering, David L.

    1975-01-01

    The spouses were counseled jointly with a cognitive approach and separately with behavior modification techniques for the husband's obesity and the wife's lack of affective verbalization. After eight weeks, the husband had lost thirty pounds and the wife's affective language had increased markedly. (Author)

  4. A NEW "PROCESSABLE" POLYACETYLENE MODIFICATION

    OpenAIRE

    Hocker, J.; Schneider, G.

    1983-01-01

    Using a special catalyst on the Ziegler-Natta basis and special reaction conditions it is possible to polymerize acetylene for obtaining a new polyacetylene modification with burr- or fibre-shaped particles which can be processed to foils, non-wovens and coatings.

  5. Supervisory Workbook on Behavior Modification.

    Science.gov (United States)

    Arkin, Ronald; And Others

    This workbook is designed to be used with the trainer's manual in supervisory training sessions on behavior modification of employees. This is one of four manuals prepared to aid supervisors in training disadvantaged groups using social reinforcement techniques. Related documents are available as VT 018 031-VT 018 035 in this issue. (MF)

  6. Synthesis and Modification of Clinoptilolite

    Directory of Open Access Journals (Sweden)

    Pavlina Ambrozova

    2017-07-01

    Full Text Available Clinoptilolite is a natural mineral with exceptional physical characteristics resulting from its special crystal structure, mainstreamed into a large zeolite group called heulandites. An overall view of the research related to the synthesis, modification and application of synthetic clinoptilolite is presented. A single phase of clinoptilolite can be hydrothermally synthesized for 1–10 days in an autoclave from various silica, alumina, and alkali sources with initial Si/Al ratio from 3.0 to 5.0 at a temperature range from 120 to 195 °C. Crystallization rate and crystallinity of clinoptilolite can be improved by seeding. The modification of clinoptilolite has received noticeable attention from the research community, since modified forms have specific properties and therefore their area of application has been broadening. This paper provides a review of the use of organic compounds such as quarter alkyl ammonium, polymer, amine and inorganic species used in the modification process, discusses the processes and mechanisms of clinoptilolite modification, and identifies research gaps and new perspectives.

  7. Synthesis and Modification of Clinoptilolite.

    Science.gov (United States)

    Ambrozova, Pavlina; Kynicky, Jindrich; Urubek, Tomas; Nguyen, Vinh Dinh

    2017-07-04

    Clinoptilolite is a natural mineral with exceptional physical characteristics resulting from its special crystal structure, mainstreamed into a large zeolite group called heulandites. An overall view of the research related to the synthesis, modification and application of synthetic clinoptilolite is presented. A single phase of clinoptilolite can be hydrothermally synthesized for 1-10 days in an autoclave from various silica, alumina, and alkali sources with initial Si/Al ratio from 3.0 to 5.0 at a temperature range from 120 to 195 °C. Crystallization rate and crystallinity of clinoptilolite can be improved by seeding. The modification of clinoptilolite has received noticeable attention from the research community, since modified forms have specific properties and therefore their area of application has been broadening. This paper provides a review of the use of organic compounds such as quarter alkyl ammonium, polymer, amine and inorganic species used in the modification process, discusses the processes and mechanisms of clinoptilolite modification, and identifies research gaps and new perspectives.

  8. Medium Modification of Vector Mesons

    Energy Technology Data Exchange (ETDEWEB)

    Chaden Djalali, Michael Paolone, Dennis Weygand, Michael H. Wood, Rakhsha Nasseripour

    2011-03-01

    The theory of the strong interaction, Quantum Chromodynamics (QCD), has been remarkably successful in describing high-energy and short-distance-scale experiments involving quarks and gluons. However, applying QCD to low energy and large-distance scale experiments has been a major challenge. Various QCD-inspired models predict a partial restoration of chiral symmetry in nuclear matter with modifications of the properties of hadrons from their free-space values. Measurable changes such as a shift in mass and/or a change of width are predicted at normal nuclear density. Photoproduction of vector mesons off nuclei have been performed at different laboratories. The properties of the ρ, ω and φ mesons are investigated either directly by measuring their mass spectra or indirectly through transparency ratios. The latest results regarding medium modifications of the vector mesons in the nuclear medium will be discussed.

  9. Posttranslational modification and quality control.

    Science.gov (United States)

    Wang, Xuejun; Pattison, J Scott; Su, Huabo

    2013-01-18

    Protein quality control functions to minimize the level and toxicity of misfolded proteins in the cell. Protein quality control is performed by intricate collaboration among chaperones and target protein degradation. The latter is performed primarily by the ubiquitin-proteasome system and perhaps autophagy. Terminally misfolded proteins that are not timely removed tend to form aggregates. Their clearance requires macroautophagy. Macroautophagy serves in intracellular quality control also by selectively segregating defective organelles (eg, mitochondria) and targeting them for degradation by the lysosome. Inadequate protein quality control is observed in a large subset of failing human hearts with a variety of causes, and its pathogenic role has been experimentally demonstrated. Multiple posttranslational modifications can occur to substrate proteins and protein quality control machineries, promoting or hindering the removal of the misfolded proteins. This article highlights recent advances in posttranslational modification-mediated regulation of intracellular quality control mechanisms and its known involvement in cardiac pathology.

  10. Laser surface modification and adhesion

    CERN Document Server

    Mittal, K L

    2014-01-01

    The book provides a unique overview on laser techniques and applications for the purpose of improving adhesion by altering surface chemistry and topography/morphology of the substrate. It details laser surface modification techniques for a wide range of industrially relevant materials (plastics, metals, ceramics, composites) with the aim to improve and enhance their adhesion to other materials. The joining of different materials is of critical importance in the fabrication of many and varied products.

  11. Synthesis and Modification of Clinoptilolite

    OpenAIRE

    Pavlina Ambrozova; Jindrich Kynicky; Tomas Urubek; Vinh Dinh Nguyen

    2017-01-01

    Clinoptilolite is a natural mineral with exceptional physical characteristics resulting from its special crystal structure, mainstreamed into a large zeolite group called heulandites. An overall view of the research related to the synthesis, modification and application of synthetic clinoptilolite is presented. A single phase of clinoptilolite can be hydrothermally synthesized for 1–10 days in an autoclave from various silica, alumina, and alkali sources with initial Si/Al ratio from 3.0 to 5...

  12. [Advances in genetic modification technologies].

    Science.gov (United States)

    Zhang, Baixue; Sun, Qixin; Li, Haifeng

    2015-08-01

    Genetic modification technology is a new molecular tool for targeted genome modification. It includes zinc finger nucleases (ZFN) technology, transcription activator-like effector nucleases (TALEN) technology and clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) (CRISPR-Cas) nucleases technology. All of these nucleases create DNA double-strand breaks (DSB) at chromosomal targeted sites and induce cell endogenous mechanisms that are primarily repaired by the non-homologous end joining (NHEJ) or homologous recombination (HR) pathway, resulting in targeted endogenous gene knock-out or exogenous gene insertion. In recent years, genetic modification technologies have been successfully applied to bacteria, yeast, human cells, fruit fly, zebra fish, mouse, rat, livestock, cynomolgus monkey, Arabidopsis, rice, tobacco, maize, sorghum, wheat, barley and other organisms, showing its enormous advantage in gene editing field. Especially, the newly developed CRISPR-Cas9 system arose more attention because of its low cost, high effectiveness, simplicity and easiness. We reviewed the principles and the latest research progress of these three technologies, as well as prospect of future research and applications.

  13. Chromatin proteins and modifications as drug targets

    DEFF Research Database (Denmark)

    Helin, Kristian; Dhanak, Dashyant

    2013-01-01

    A plethora of groundbreaking studies have demonstrated the importance of chromatin-associated proteins and post-translational modifications of histones, proteins and DNA (so-called epigenetic modifications) for transcriptional control and normal development. Disruption of epigenetic control...

  14. Energy conservation potential of surface modification technologies

    Energy Technology Data Exchange (ETDEWEB)

    Le, H.K.; Horne, D.M.; Silberglitt, R.S.

    1985-09-01

    This report assesses the energy conservation impact of surface modification technologies on the metalworking industries. The energy conservation impact of surface modification technologies on the metalworking industries is assessed by estimating their friction and wear tribological sinks and the subsequent reduction in these sinks when surface modified tools are used. Ion implantation, coatings, and laser and electron beam surface modifications are considered.

  15. Epigenetic modifications and diabetic nephropathy

    Directory of Open Access Journals (Sweden)

    Marpadga A. Reddy

    2012-09-01

    Full Text Available Diabetic nephropathy (DN is a major complication associated with both type 1 and type 2 diabetes, and a leading cause of end-stage renal disease. Conventional therapeutic strategies are not fully efficacious in the treatment of DN, suggesting an incomplete understanding of the gene regulation mechanisms involved in its pathogenesis. Furthermore, evidence from clinical trials has demonstrated a “metabolic memory” of prior exposure to hyperglycemia that continues to persist despite subsequent glycemic control. This remains a major challenge in the treatment of DN and other vascular complications. Epigenetic mechanisms such as DNA methylation, nucleosomal histone modifications, and noncoding RNAs control gene expression through regulation of chromatin structure and function and post-transcriptional mechanisms without altering the underlying DNA sequence. Emerging evidence indicates that multiple factors involved in the etiology of diabetes can alter epigenetic mechanisms and regulate the susceptibility to diabetes complications. Recent studies have demonstrated the involvement of histone lysine methylation in the regulation of key fibrotic and inflammatory genes related to diabetes complications including DN. Interestingly, histone lysine methylation persisted in vascular cells even after withdrawal from the diabetic milieu, demonstrating a potential role of epigenetic modifications in metabolic memory. Rapid advances in high-throughput technologies in the fields of genomics and epigenomics can lead to the identification of genome-wide alterations in key epigenetic modifications in vascular and renal cells in diabetes. Altogether, these findings can lead to the identification of potential predictive biomarkers and development of novel epigenetic therapies for diabetes and its associated complications.

  16. MAP kinases and histone modification

    Institute of Scientific and Technical Information of China (English)

    Tamaki Suganuma; Jerry L. Workman

    2012-01-01

    Signal transduction pathways alter the gene expression program in response to extracellular or intracellular cues.Mitogen-activated protein kinases (MAPKs) govern numerous cellular processes including cell growth,stress response,apoptosis,and differentiation.In the past decade,MAPKs have been shown to regulate the transcription machinery and associate with chromatin-modifying complexes.Moreover,recent studies demonstrate that several MAPKs bind directly to chromatin at target genes.This review highlights the recent discoveries of MAPK signaling in regard to histone modifications and chromatin regulation.Evidence suggesting that further unknown mechanisms integrate signal transduction with chromatin biology is discussed.

  17. Mechanochemical modification of natural rubber

    Science.gov (United States)

    Mikhaylov, I. A.; Sukhareva, K. V.; Andriasyan, Yu. O.; Popov, A. A.; Vorontsov, N. V.

    2016-11-01

    Thermomechanochemical changes of SVR 3L natural rubber after the treatment in the internal rubber mixer in the self-heating mode were studied. The effect of the molecular mass and content of the gel fraction of natural rubber is shown. Properties of rubber compounds and vulcanized rubber are presented. Taking into account modern requirements, a new alternative technology of obtaining halogenated elastomers based on the solid-phase (mechanochemical) halide modification is created. New halogen-containing natural rubber produced by this technology proves themselves in the conditions of rubber production. New fluorinated natural rubber produced by this technology proves themselves in the conditions of rubber production.

  18. Chemical Modification of Food Proteins

    Institute of Scientific and Technical Information of China (English)

    Allaoua Achouri; Wang Zhang; Xu Shiying

    1999-01-01

    Acylation has been shown to be an effective tool for improving surface functional properties of plant proteins.Soy bean protein has been extensively modified through chemical and enzymatic treatments. Their effectiveness lies in their high nutritional value and low cost, which promote their use as ingredients for the formulation of food products.This paper reports a complete review of chemical modification of various proteins from plant and animal sources. The nutritive and toxicological aspects through in vitro and in vivo tests are also described.

  19. Modifications to Ivor Lewis esophagectomy.

    Science.gov (United States)

    David, Elizabeth A; Marshall, M Blair

    2010-11-01

    The surgical approach to esophagectomy is variable. A number of factors are considered when determining the optimal approach to esophagectomy: location and extent of disease, fibrosis, additional patient factors and surgeon preference. One of the disadvantages to some approaches is the need for a change in position, which increases operative time. Also, because typically the abdomen is initially explored, patients may later be deemed unresectable at thoracotomy. We describe time saving modifications to the standard Ivor Lewis esophagectomy that eliminate the need for repositioning and facilitate a stapled end-to-end anastomosis.

  20. [Bronchial morphologic modification in asthma].

    Science.gov (United States)

    Cataldo, D; Louis, R; Godon, A; Munaut, C; Noël, A; Foidart, J M; Bartsch, P

    2000-07-01

    Asthma is an inflammatory disease of the airways clinically characterised by recurrent bronchial obstructions at least partially reversible. Recent epidemiologic data suggest that asthmatics have an increased rate of decrease of their expiratory volumes during life. This irreversible lung function impairment is associated with fundamental structural changes of the bronchial wall in terms of conjunctive tissue and smooth muscle composition. We describe these changes and explore the different mechanisms proposed to explain these structural modifications. We also review their consequences in terms of bronchial physiology and their potential influence on bronchial hyperresponsiveness.

  1. DNA Modifications and Alzheimer's Disease.

    Science.gov (United States)

    Smith, Rebecca G; Lunnon, Katie

    2017-01-01

    Alzheimer's disease (AD) is a complex neurodegenerative disease, affecting millions of people worldwide. While a number of studies have focused on identifying genetic variants that contribute to the development and progression of late-onset AD, the majority of these only have a relatively small effect size. There are also a number of other risk factors, for example, age, gender, and other comorbidities; however, how these influence disease risk is not known. Therefore, in recent years, research has begun to investigate epigenetic mechanisms for a potential role in disease etiology. In this chapter, we discuss the current state of play for research into DNA modifications in AD, the most well studied being 5-methylcytosine (5-mC). We describe the earlier studies of candidate genes and global measures of DNA modifications in human AD samples, in addition to studies in mouse models of AD. We focus on recent epigenome-wide association studies (EWAS) in human AD, using microarray technology, examining a number of key study design issues pertinent to such studies. Finally, we discuss how new technological advances could further progress the research field.

  2. Realities of craniofacial growth modification.

    Science.gov (United States)

    Kluemper, G T; Spalding, P M

    2001-03-01

    Facial growth modification can be an effective method of resolving skeletal discrepancies. There still is much controversy regarding our understanding of the nature and extent of skeletal orthopedic change possible in individual patients and the most effective appliances and timing of such treatment. In the treatment of class II patients, growth modification can lead to an improvement, if not complete correction of the class II malocclusion. Although two-phase treatment with an early first prepubertal phase can be effective, a later single-phase approach during early puberty seems to be equally effective. Certainly, before surgical correction of the mild to moderate skeletal class II problem in a growing patient is considered, an orthopedic phase of treatment prior to the pubertal growth spurt is an appropriate first step. Skeletal class III patients with a maxillary deficiency stand to gain significant benefits from early orthopedic treatment. However, such therapy may produce more favorable changes for older children and adolescents than previously thought. Nevertheless, orthopedic correction of the mild to moderate skeletal class III should be accompanied by regular progress evaluations to avoid creating significant dental compensations in the face with little skeletal change that ultimately requires surgery anyway. Skeletal class III patients with mandibular excess and/or vertical excess are poor candidates for growth modification. Orthopedic palatal expansion appears to be effective and stable at any time prior to late puberty, a stage of development when ossification of the maxillary sutures is more advanced. Consequently, the timing for expansion may be better determined by the specific needs of each patient. A functional shift resulting from a crossbite is optimally corrected early, so that asymmetric growth of the mandible can be reduced or even prevented. Postpubertal orthopedic expansion is likely to result in bone bending, which will reverse itself over

  3. Germline modification of domestic animals.

    Science.gov (United States)

    Tang, L; González, R; Dobrinski, I

    2015-01-01

    Genetically-modified domestic animal models are of increasing significance in biomedical research and agriculture. As authentic ES cells derived from domestic animals are not yet available, the prevailing approaches for engineering genetic modifications in those animals are pronuclear microinjection and somatic cell nuclear transfer (SCNT, also known as cloning). Both pronuclear microinjection and SCNT are inefficient, costly, and time-consuming. In animals produced by pronuclear microinjection, the exogenous transgene is usually inserted randomly into the genome, which results in highly variable expression patterns and levels in different founders. Therefore, significant efforts are required to generate and screen multiple founders to obtain animals with optimal transgene expression. For SCNT, specific genetic modifications (both gain-of-function and loss-of-function) can be engineered and carefully selected in the somatic cell nucleus before nuclear transfer. SCNT has been used to generate a variety of genetically modified animals such as goats, pigs, sheep and cattle; however, animals resulting from SCNT frequently suffer from developmental abnormalities associated with incomplete nuclear reprogramming. Other strategies to generate genetically-modified animals rely on the use of the spermatozoon as a natural vector to introduce genetic material into the female gamete. This sperm mediated DNA transfer (SMGT) combined with intracytoplasmatic sperm injection (ICSI) has relatively high efficiency and allows the insertion of large DNA fragments, which, in turn, enhance proper gene expression. An approach currently being developed to complement SCNT for producing genetically modified animals is germ cell transplantation using genetically modified male germline stem cells (GSCs). This approach relies on the ability of GSCs that are genetically modified in vitro to colonize the recipient testis and produce donor derived sperm upon transplantation. As the genetic change

  4. Surface Modification for Microreactor Fabrication

    Directory of Open Access Journals (Sweden)

    Wladyslaw Torbicz

    2006-04-01

    Full Text Available In this paper, methods of surface modification of different supports, i.e. glass andpolymeric beads for enzyme immobilisation are described. The developed method ofenzyme immobilisation is based on Schiff’s base formation between the amino groups onthe enzyme surface and the aldehyde groups on the chemically modified surface of thesupports. The surface of silicon modified by APTS and GOPS with immobilised enzymewas characterised by atomic force microscopy (AFM, time-of-flight secondary ion massspectroscopy (ToF-SIMS and infrared spectroscopy (FTIR. The supports withimmobilised enzyme (urease were also tested in combination with microreactors fabricatedin silicon and Perspex, operating in a flow-through system. For microreactors filled withurease immobilised on glass beads (Sigma and on polymeric beads (PAN, a very high andstable signal (pH change was obtained. The developed method of urease immobilisationcan be stated to be very effective.

  5. Soy protein modification: A review

    Directory of Open Access Journals (Sweden)

    Barać Miroljub B.

    2004-01-01

    Full Text Available Soy protein products such as flour, concentrates and isolates are used in food formulation because of their functionality, nutritional value and low cost. To obtain their optimal nutritive and functional properties as well as desirable flavor different treatments are used. Soybean proteins can be modified by physical, chemical and enzymatic treatments. Different thermal treatments are most commonly used, while the most appropriate way of modifying soy proteins from the standpoint of safety is their limited proteolysis. These treatments cause physical and chemical changes that affect their functional properties. This review discusses three principal methods used for modification of soy protein products, their effects on dominant soy protein properties and some biologically active compounds.

  6. Surface modification of cellulose nanocrystals

    Institute of Scientific and Technical Information of China (English)

    WANG Neng; DING Enyong; CHENG Rongshi

    2007-01-01

    In order to improve the dispersibility of cellulose nanocrystal(CNC) particles,three difierent grafted reactions of acetylation,hydroxyethylation and hydroxypropylation were introduced to modify the CNC surface.The main advantages of these methods were the simple and easily controlled reaction conditions,and the dispersibility of the resulting products was distinctly improved.The properties of the modified CNC were characterized by means of Fourier transform infrared spectroscopy(FT-IR),13 C nuclear magnetic resonance(NMR),transmission electron microscopy(TEM)and thermogravimetric analyses(TGA).The results indicated mat after desiccation,the modification products could be dispersed again in the proper solvents by ultrasonic treatments,and the diameter of their particles had no obvious changes.However,their thermal degradation behaviors were quite different.The initial decomposition temperature of the modified products via hydroxyethylation or hydroxypropylation was lower than that of modified products via acetylation.

  7. Review of Rule Modification in Sport

    OpenAIRE

    Arias, Jose L.; Francisco M. Argudo; Alonso, Jose I.

    2011-01-01

    The goal of this qualitative review was to analyze the state of the bibliography about rule modification in sport. In the literature reviewed, there are few studies of rule modification and related aspects. Most studies omit mentioning the purpose of the modifications, but they do refer to the goals of their analysis (improving players' performance, attracting spectators and athletes, attending to commercial pressure, adapting the sport to children's needs and interests, preventing injuries)....

  8. Climate modification directed by control theory

    CERN Document Server

    Liang, Wang

    2008-01-01

    Climate modification measures to counteract global warming receive some more new attentions in these years. Most current researches only discuss the impact of these measures to climate, but how to design such a climate regulator is still unknown. This paper shows the control theory could give the systematic direction for climate modification. But the control analyzing also reveals that climate modifications should only be regarded as a last-ditch measure.

  9. Proteomic analysis of post-translational modifications

    DEFF Research Database (Denmark)

    Mann, Matthias; Jensen, Ole N

    2003-01-01

    Post-translational modifications modulate the activity of most eukaryote proteins. Analysis of these modifications presents formidable challenges but their determination generates indispensable insight into biological function. Strategies developed to characterize individual proteins are now...... mass spectrometric peptide sequencing and analysis technologies hold tremendous potential. Finally, stable isotope labeling strategies in combination with mass spectrometry have been applied successfully to study the dynamics of modifications....

  10. Chemical Modifications of Starch: Microwave Effect

    OpenAIRE

    Kamila Lewicka; Przemysław Siemion; Piotr Kurcok

    2015-01-01

    This paper presents basic methods of starch chemical modification, the effect of microwave radiation on the modification process, and the physicochemical properties of starch. It has been shown that the modifications contribute to improvement of the material performance and likewise to significant improvement of its mechanical properties. As a result, more and more extensive use of starch is possible in various industries. In addition, methods of oxidized starch and starch esters preparation ...

  11. Epigenetic Modifications and Plant Hormone Action.

    Science.gov (United States)

    Yamamuro, Chizuko; Zhu, Jian-Kang; Yang, Zhenbiao

    2016-01-04

    The action of phytohormones in plants requires the spatiotemporal regulation of their accumulation and responses at various levels. Recent studies reveal an emerging relationship between the function of phytohormones and epigenetic modifications. In particular, evidence suggests that auxin biosynthesis, transport, and signal transduction is modulated by microRNAs and epigenetic factors such as histone modification, chromatin remodeling, and DNA methylation. Furthermore, some phytohormones have been shown to affect epigenetic modifications. These findings are shedding light on the mode of action of phytohormones and are opening up a new avenue of research on phytohormones as well as on the mechanisms regulating epigenetic modifications.

  12. Methodological guidelines for developing accident modification functions

    DEFF Research Database (Denmark)

    Elvik, Rune

    2015-01-01

    This paper proposes methodological guidelines for developing accident modification functions. An accident modification function is a mathematical function describing systematic variation in the effects of road safety measures. The paper describes ten guidelines. An example is given of how to use...... limitations in developing accident modification functions are the small number of good evaluation studies and the often huge variation in estimates of effect. It is therefore still not possible to develop accident modification functions for very many road safety measures. © 2015 Elsevier Ltd. All rights...

  13. Chemical biology approaches for studying posttranslational modifications.

    Science.gov (United States)

    Yang, Aerin; Cho, Kyukwang; Park, Hee-Sung

    2017-09-13

    Posttranslational modification (PTM) is a key mechanism for regulating diverse protein functions, and thus critically affects many essential biological processes. Critical for systematic study of the effects of PTMs is the ability to obtain recombinant proteins with defined and homogenous modifications. To this end, various synthetic and chemical biology approaches, including genetic code expansion and protein chemical modification methods, have been developed. These methods have proven effective for generating site-specific authentic modifications or structural mimics, and have demonstrated their value for in vitro and in vivo functional studies of diverse PTMs. This review will discuss recent advances in chemical biology strategies and their application to various PTM studies.

  14. Chemical Modifications of Starch: Microwave Effect

    Directory of Open Access Journals (Sweden)

    Kamila Lewicka

    2015-01-01

    Full Text Available This paper presents basic methods of starch chemical modification, the effect of microwave radiation on the modification process, and the physicochemical properties of starch. It has been shown that the modifications contribute to improvement of the material performance and likewise to significant improvement of its mechanical properties. As a result, more and more extensive use of starch is possible in various industries. In addition, methods of oxidized starch and starch esters preparation are discussed. Properties of microwave radiation and its impact on starch (with particular regard to modifications described in literature are characterized.

  15. Bioactive Surface Modification of Hydroxyapatite

    Directory of Open Access Journals (Sweden)

    Yasuhiko Abe

    2013-01-01

    Full Text Available The purpose of this study was to establish an acid-etching procedure for altering the Ca/P ratio of the nanostructured surface of hydroxyapatite (HAP by using surface chemical and morphological analyses (XPS, XRD, SEM, surface roughness, and wettability and to evaluate the in vitro response of osteoblast-like cells (MC3T3-E1 cells to the modified surfaces. This study utilized HAP and HAP treated with 10%, 20%, 30%, 40%, 50%, or 60% phosphoric acid solution for 10 minutes at 25°C, followed by rinsing 3 times with ultrapure water. The 30% phosphoric acid etching process that provided a Ca/P ratio of 1.50, without destruction of the grain boundary of HAP, was selected as a surface-modification procedure. Additionally, HAP treated by the 30% phosphoric acid etching process was stored under dry conditions at 25°C for 12 hours, and the Ca/P ratio approximated to 1.00 accidentally. The initial adhesion, proliferation, and differentiation (alkaline phosphatase (ALP activity and relative mRNA level for ALP of MC3T3-E1 cells on the modified surfaces were significantly promoted (P<0.05 and 0.01. These findings show that the 30% phosphoric acid etching process for the nanostructured HAP surface can alter the Ca/P ratio effectively and may accelerate the initial adhesion, proliferation, and differentiation of MC3T3-E1 cells.

  16. 38 CFR 36.4315 - Loan modifications.

    Science.gov (United States)

    2010-07-01

    ... back from the modification. (j) This section does not create a right of a borrower to have a loan... 38 Pensions, Bonuses, and Veterans' Relief 2 2010-07-01 2010-07-01 false Loan modifications. 36...) LOAN GUARANTY Guaranty or Insurance of Loans to Veterans With Electronic Reporting § 36.4315...

  17. Histone modifications: Cycling with chromosomal replication

    DEFF Research Database (Denmark)

    Thon, Genevieve

    2008-01-01

    Histone modifications tend to be lost during chromosome duplication. Several recent studies suggest that the RNA interference pathway becomes active during the weakened transcriptional repression occurring at centromeres in S phase, resulting in the re-establishment of histone modifications that ...

  18. Characterization of Surface Modification of Polyethersulfone Membrane

    Science.gov (United States)

    Surface modification of polyethersulfone (PES) membrane surface using UV/ozone-treated grafting and interfacial polymerization on membrane surface was investigated in order to improve the resistance of membrane surface to protein adsorption. These methods of surface modification were compared in te...

  19. Electrochemical modification process of anodic alumina membrane

    Institute of Scientific and Technical Information of China (English)

    YU Mei; LIU Jian-hua; LI Song-mei

    2006-01-01

    The modification procedure of anodic alumina membrane(AAM) was studied. The AAM structure after modification was characterized by nickel nanowires prepared in AAM. Scanning electron microscopy was used to characterize the topography and structure properties of the AAM and nickel nanowires. The transformation of the current during the voltage reduction was studied. The mechanism of current and structure change during modification was discussed. The results show that a root structure produces after the AAM modification. The length of the root structure depends on the velocity of the voltage reduction. Slow voltage reduction leads to a large length of the root structure,otherwise,a short length of the root structure. At the end of the modification,the barrier layer is thin enough to be passed by electrons. Hence,the direct electrodeposition of one-dimensional nanowires can be carried out on the AAM with barrier layer and aluminum matrix successfully without any other treatments.

  20. Modification Semantics in Now-Relative Databases

    DEFF Research Database (Denmark)

    Torp, Kristian; Jensen, Christian Søndergaard; Snodgrass, R. T.

    2004-01-01

    Most real-world databases record time-varying information. In such databases, the notion of ??the current time,?? or NOW, occurs naturally and prominently. For example, when capturing the past states of a relation using begin and end time columns, tuples that are part of the current state have some...... past time as their begin time and NOW as their end time. While the semantics of such variable databases has been described in detail and is well understood, the modification of variable databases remains unexplored. This paper defines the semantics of modifications involving the variable NOW. More...... specifically,  the problems with modifications in the presence of NOW are explored, illustrating that the main problems are with modifications of tuples that reach into the future. The paper defines the semantics of modifications?including insertions, deletions, and updates?of databases without NOW, with NOW...

  1. REVIEW OF RULE MODIFICATION IN SPORT

    Directory of Open Access Journals (Sweden)

    Jose L. Arias

    2011-03-01

    Full Text Available The goal of this qualitative review was to analyze the state of the bibliography about rule modification in sport. In the literature reviewed, there are few studies of rule modification and related aspects. Most studies omit mentioning the purpose of the modifications, but they do refer to the goals of their analysis (improving players' performance, attracting spectators and athletes, attending to commercial pressure, adapting the sport to children's needs and interests, preventing injuries. Eighty percent of the studies did not report the outcome of the previous modifications they analyzed. More than half of the studies (60% achieved the proposed goals. Nearly two-thirds (63.83% analyzed the effect of rule modification on game actions occurring during the game or through a test. Most of the studies (91.5% did not consult the participants. Three-fourths of the studies (74.46% examined the effect of rule modification without any knowledge of a previous analysis or without any previous analysis, and 74.47% studied rule modification related to internal logic. Modifications to be introduced in a sport should be analyzed through a reflective process before their final introduction. The following points should be considered: establishing goals, respecting the basic rules without modifying them, becoming familiar with players' and coaches' opinions, determining the effect of the modification on a wide spectrum of variables, elaborating useful proposals for the organizations that are responsible for competitions, using more than one type of data, modifying the internal logic and, preferably, the functional rules, and following some basic stages to consolidate rule modification.

  2. Modifications to the INSPECT model

    Energy Technology Data Exchange (ETDEWEB)

    Dickinson, S. [AEA Technology, Winfrith (United Kingdom); Sims, H.E. [AEA Technology, Harwell (United Kingdom)

    1996-12-01

    The prediction of iodine behaviour in the containment of a PWR following a loss of coolant accident requires a reliable model of the chemistry of iodine in aqueous solution. The INSPECT model, which was developed several years ago, contains a large number of the relevant chemical reactions of iodine and water radiation chemistry. Since the reactions set was first assembled, however, new data on rate constants and mechanisms have become available. In addition, the application of the model to various small-scale experiments has revealed problems in the modelling of some reactions, leading to an under-prediction of the iodine volatility at high pH, although the experiments have demonstrated that the high pH volatility remains satisfactory low. This paper describes the modifications which have been made to the INSPECT model to take account of new data and to improve the modelling where appropriate. The main changes which have been made to the reaction set are as follows: - The rate constants and activation energies for the reactions describing the radiolysis of water have been updated in with recent assessments, and the temperature dependence of the G value for the primary species have been accordance taken into account, - The mechanism and rates of I{sub 2} hydrolysis have been modified in accordance with the latest assessments of this reaction, - The mechanism for the reaction of I{sub 2} with H{sub 2}O{sub 2} has been changed to a form which produces a more realistic pH dependence under neutral and alkaline conditions, - An addition mechanism for the disproportionation of the O{sub 2} ion has been included, reflecting experimental observations that this reaction has a significant first-order component under all but the purest conditions, - Atomic I is treated as a volatile species, with a partition coefficient of 1.9 at 298 K. (Abstract Truncated)

  3. Physical modification of polyetheretherketone for orthopedic implants

    Science.gov (United States)

    Du, Ya-Wei; Zhang, Li-Nan; Hou, Zeng-Tao; Ye, Xin; Gu, Hong-Sheng; Yan, Guo-Ping; Shang, Peng

    2014-12-01

    Polyetheretherketone (PEEK) is regarded as one of the most potential candidates for replacing current implant applications. To obtain good bone-implant interfaces, many modification methods have been developed to enable PEEK and PEEK-based composites from bio-inert to bioactive. Among them, physical methods have aroused significant attention and been widely used to modify PEEK for orthopedic implants. This review summarizes current physical modification techniques of PEEK for orthopedic applications, which include composite strategies, surface coating methods and irradiation treatments. The positive consequences of those modification methods will encourage continuing investigations and stimulate the wide range of applications of PEEK-based implants in orthopedics.

  4. [Behavior modification therapy: application in renal nursing].

    Science.gov (United States)

    Wang, Lu-Mei; Hung, Shih-Yuan; Chiou, Chou-Ping

    2011-04-01

    Kidney patients often suffer numerous comorbidities (e.g., hyperphosphatemia, renal osteodystrophy, and bodyweight gain induced by fluid overload or hypertension), which often impart discomfort and a significantly reduced quality of life. The literatures points to the efficacy of behavior modification therapy in changing inappropriate self-health care behavior in order to improve comorbidity symptoms. This article introduces behavior modification theory and its application as well as clinical strategies related to kidney patient care. The authors hope this article will promote knowledge of behavior modification theory among healthcare professionals and facilitate to the application of this theory in clinical practice to improve nursing care.

  5. Epigenetic Modifications: Therapeutic Potential in Cancer

    Directory of Open Access Journals (Sweden)

    Manisha Sachan

    2015-08-01

    Full Text Available Epigenetic modifications and alterations in chromatin structure and function contribute to the cumulative changes observed as normal cells undergo malignant transformation. These modifications and enzymes (DNA methyltransferases, histone deacetylases, histone methyltransferases, and demethylases related to them have been deeply studied to develop new drugs, epigenome-targeted therapies and new diagnostic tools. Epigenetic modifiers aim to restore normal epigenetic modification patterns through the inhibition of epigenetic modifier enzymes. Four of them (azacitidine, decitabine, vorinostat and romidepsin are approved by the U.S. Food and Drug Administration. This article provides an overview about the known functional roles of epigenetic enzymes in cancer development.

  6. Nanomaterials Applied in Asphalt Modification: A Review

    Institute of Scientific and Technical Information of China (English)

    Changqing Fang; Ruien Yu; Shaolong Liu; Yan Li

    2013-01-01

    Nanotechnology has been gradually penetrated into the field of asphalt modification.Seemingly magic effects of nanomaterials have now been brought to improve the performance of asphalt.To demonstrate many of the prospective applications,researchers have conducted a series of positive and effective efforts dealing with the preparation of modified asphalt to demonstrate the mechanism of modification and the resultant improvement in performance.In this review,various nanomaterials used in asphalt modification are initially presented,followed by the methods employed to modify the asphalt with these materials and finally the effects of nanomaterials on the performance of base asphalt are presented and the modification mechanisms are discussed.Based on the current research results,the influence of preparation process parameters on the compatibility of every phase in the modified asphalt and the stability of the modified asphalt system are described.Finally,the development trend of the topic field is projected.

  7. Preventing cancer through lifestyle modification: An essential ...

    African Journals Online (AJOL)

    Preventing cancer through lifestyle modification: An essential strategy for primary ... The ten most common cancers include Cervical, Breast, Prostate, Liver, ... viral, bacterial and helminthic infections; tobacco use, poor dietary habits, obesity, ...

  8. Diagonal chromatography to study plant protein modifications

    NARCIS (Netherlands)

    Walton, Alan; Tsiatsiani, Liana; Jacques, Silke; Stes, Elisabeth; Messens, Joris; Van Breusegem, Frank; Goormachtig, Sofie; Gevaert, Kris

    2016-01-01

    An interesting asset of diagonal chromatography, which we have introduced for contemporary proteome research, is its high versatility concerning proteomic applications. Indeed, the peptide modification or sorting step that is required between consecutive peptide separations can easily be altered and

  9. Similarity Arguments in the Genetic Modification Debate

    DEFF Research Database (Denmark)

    Christiansen, Andreas

    2017-01-01

    In the ethical debate on genetic modification (GM), it is common to encounter the claim that some anti-GM argument would also apply an established, ethically accepted technology, and that the anti-GM argument is therefore unsuccessful. The paper discusses whether this argumentative strategy...... transferability of reasons from one case to another; and (iii) it runs the risk of equivocations, especially in cases where the anti-genetic-modification argument relies on gradable features. The paper then shows how these issues play out in three specific Similarity Arguments that can be found in the literature....... Finally, the paper discusses what conclusions we can draw from the fact that genetic modification and established technologies are similar for the ethical status of genetic modification....

  10. Assessment of posttranslational modification of mitochondrial proteins.

    Science.gov (United States)

    Ande, Sudharsana R; Padilla-Meier, G Pauline; Mishra, Suresh

    2015-01-01

    Mitochondria play vital roles in the maintenance of cellular homeostasis. They are a storehouse of cellular energy and antioxidative enzymes. Because of its immense role and function in the development of an organism, this organelle is required for the survival. Defects in mitochondrial proteins lead to complex mitochondrial disorders and heterogeneous diseases such as cancer, type 2 diabetes, and cardiovascular and neurodegenerative diseases. It is widely known in the literature that some of the mitochondrial proteins are regulated by posttranslational modifications. Hence, designing methods to assess these modifications in mitochondria will be an important way to study the regulatory roles of mitochondrial proteins in greater detail. In this chapter, we outlined procedures to isolate mitochondria from cells and separate the mitochondrial proteins by two-dimensional gel electrophoresis and identify the different posttranslational modifications in them by using antibodies specific to each posttranslational modification.

  11. Post-Translational Modifications of TRP Channels

    Directory of Open Access Journals (Sweden)

    Olaf Voolstra

    2014-04-01

    Full Text Available Transient receptor potential (TRP channels constitute an ancient family of cation channels that have been found in many eukaryotic organisms from yeast to human. TRP channels exert a multitude of physiological functions ranging from Ca2+ homeostasis in the kidney to pain reception and vision. These channels are activated by a wide range of stimuli and undergo covalent post-translational modifications that affect and modulate their subcellular targeting, their biophysical properties, or channel gating. These modifications include N-linked glycosylation, protein phosphorylation, and covalent attachment of chemicals that reversibly bind to specific cysteine residues. The latter modification represents an unusual activation mechanism of ligand-gated ion channels that is in contrast to the lock-and-key paradigm of receptor activation by its agonists. In this review, we summarize the post-translational modifications identified on TRP channels and, when available, explain their physiological role.

  12. Surface Modification of Water Purification Membranes.

    Science.gov (United States)

    Miller, Daniel J; Dreyer, Daniel R; Bielawski, Christopher W; Paul, Donald R; Freeman, Benny D

    2017-04-18

    Polymeric membranes are an energy-efficient means of purifying water, but they suffer from fouling during filtration. Modification of the membrane surface is one route to mitigating membrane fouling, as it helps to maintain high levels of water productivity. Here, a series of common techniques for modification of the membrane surface are reviewed, including surface coating, grafting, and various treatment techniques such as chemical treatment, UV irradiation, and plasma treatment. Historical background on membrane development and surface modification is also provided. Finally, polydopamine, an emerging material that can be easily deposited onto a wide variety of substrates, is discussed within the context of membrane modification. A brief summary of the chemistry of polydopamine, particularly as it may pertain to membrane development, is also described. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Covalent Surface Modifications of Carbon Nanotubes.

    Energy Technology Data Exchange (ETDEWEB)

    Pavia Sanders, Adriana [Sandia National Lab. (SNL-CA), Livermore, CA (United States); O' Bryan, Greg [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

    2017-07-01

    A report meant to document the chemistries investigated by the author for covalent surface modification of CNTs. Oxidation, cycloaddition, and radical reactions were explored to determine their success at covalently altering the CNT surface. Characterization through infrared spectroscopy, Raman spectroscopy, and thermo gravimetric analysis was performed in order to determine the success of the chemistries employed. This report is not exhaustive and was performed for CNT surface modification exploration as it pertains to the "Next Gen" project.

  14. DNA Methylation: Bisulphite Modification and Analysis

    OpenAIRE

    Patterson, Kate; Molloy, Laura; Qu, Wenjia; Clark, Susan

    2011-01-01

    Epigenetics describes the heritable changes in gene function that occur independently to the DNA sequence. The molecular basis of epigenetic gene regulation is complex, but essentially involves modifications to the DNA itself or the proteins with which DNA associates. The predominant epigenetic modification of DNA in mammalian genomes is methylation of cytosine nucleotides (5-MeC). DNA methylation provides instruction to gene expression machinery as to where and when the gene should be expres...

  15. Post-translational modifications in neurodegeneration

    Directory of Open Access Journals (Sweden)

    Federico Benetti

    2015-12-01

    Full Text Available Post-translational modifications increase proteome functionality for managing all aspects of normal cell biology. They are based on the covalent attachment of functional groups, leading to phosphorylation, acetylation, glycosylation, acylation, ubiquitination, SUMOylation and oxidation of protein targets. Post-translational modifications occur at any step of protein life cycle, modulating in time and space protein folding, subcellular localization and activity. Aberrant post-translational modifications of one or more culprit proteins may lead to neurodegeneration, as shown in paradigmatic neurological disorders such as Alzheimer’s, Parkinson’s and prion diseases. In this review, we report the most important post-translational modifications found in neurodegenerative disorders, illustrating the pathophysiological mechanisms in which they are involved. This work highlights the lack of a global framework of post-translational modifications in terms of complexity and regulation. Therefore, in the next future many efforts are required to describe the interplay existing between post-translational modifications and their combinatorial patterns on protein targets.

  16. Chromatin Modifications Associated With Diabetes and Obesity.

    Science.gov (United States)

    Schones, Dustin E; Leung, Amy; Natarajan, Rama

    2015-07-01

    The incidence of obesity across the globe has doubled over the past several decades, leading to escalating rates of diabetes mellitus, cardiovascular disease, and other complications. Given this dramatic rise in disease incidence, understanding the cause of these diseases is therefore of paramount importance. Metabolic diseases, such as obesity and diabetes mellitus, result from a multitude of genetic and environmental factors. Although the genetic basis of these diseases has been extensively studied, the molecular pathways whereby environmental factors influence disease progression are only beginning to be understood. One manner by which environmental factors can contribute to disease progression is through modifications to chromatin. The highly structured packaging of the genome into the nucleus through chromatin has been shown to be fundamental to tissue-specific gene regulation. Modifications to chromatin can regulate gene expression and are involved in a myriad of biological functions, and hence, disruption of these modifications is central to many human diseases. These modifications can furthermore be epigenetic in nature, thereby contributing to prolonged disease risk. Recent work has demonstrated that modifications to chromatin are associated with the progression of both diabetes mellitus and obesity, which is the subject of this review. © 2015 American Heart Association, Inc.

  17. Identification and Interrogation of Combinatorial Histone Modifications

    Directory of Open Access Journals (Sweden)

    Kelly R Karch

    2013-12-01

    Full Text Available Histone proteins are dynamically modified to mediate a variety of cellular processes including gene transcription, DNA damage repair, and apoptosis. Regulation of these processes occurs through the recruitment of non-histone proteins to chromatin by specific combinations of histone post-translational modifications (PTMs. Mass spectrometry has emerged as an essential tool to discover and quantify histone PTMs both within and between samples in an unbiased manner. Developments in mass spectrometry that allow for characterization of large histone peptides or intact protein has made it possible to determine which modifications occur simultaneously on a single histone polypeptide. A variety of techniques from biochemistry, biophysics, and chemical biology have been employed to determine the biological relevance of discovered combinatorial codes. This review first describes advancements in the field of mass spectrometry that have facilitated histone PTM analysis and then covers notable approaches to probe the biological relevance of these modifications in their nucleosomal context.

  18. Design inspections for systems undergoing design modifications

    Energy Technology Data Exchange (ETDEWEB)

    Grimes, B.K.; Norkin, D.P.

    1987-01-01

    The US Nuclear Regulatory Commission's (NRC's) Office of Inspection and Enforcement (IE) has conducted direct inspections (integrated design inspections) of the technical aspects of the nuclear plant design process for several plants under construction. It has also evaluated independent design verification programs and engineering assurance programs at a number of other plants under construction. Many of the lessons learned from these construction phase efforts are directly applicable to operating plants undergoing major modifications. This has been confirmed by IE direct inspections of the design aspects of safety systems outage modifications, which have identified significant potential safety-related deficiencies. The paper discusses methodology for design inspections; failure to maintain up-to-date design basis documentation; and lessons learned common to plants under construction and undergoing major modifications.

  19. Proposed Modification of the Progressive Retirement Programme

    CERN Document Server

    2003-01-01

    After discussion at TREF on 29 October 2003, the Management proposes in this document a modification affecting one of the general principles of the Progressive Retirement Programme (PRP), decided by the Council in December 1996 and introduced in April 1997. This modification would authorise the Director-General, in exceptional cases, as indicated in section 3 below, and, with the staff member's consent, to cancel his or her participation in the PRP and to reinstate the staff member in his or her original contractual situation. No modifications to the Staff Rules and Regulations are required. The proposal set out in section 3 of the present document is submitted by the Management, after consultation of the Governing Board of the Pension Fund, for recommendation by the Finance Committee to the Council and for approval by the Council, to enter into force as of 1 January 2004.

  20. [Epigenic modifications associated with low benzene exposure].

    Science.gov (United States)

    Fustinoni, Silvia; Bollati, Valentina; Bertazzi, Pier Alberto

    2013-01-01

    DNA methylation, mitochondrial DNA copy number and telomeres shortening are cellular modifications associated with an increasing number of tumors, cardiovascular and aging diseases. In our studies these modifications were evaluated in subjects occupationally exposed to low levels of benzene and in the general population. In peripheral blood lymphocytes a decrease of DNA methylation with the increase of personal benzene exposure was found, both in Alu and LINE-1 repetitive elements, and in the global DNA. Telomere length shortening in subjects exposed to traffic exhausts and an increase in mitochondrial DNA copy number correlated to benzene exposure was also found. DNA methylation measured in specimen repeats collected at intervals of 8 years decreased more markedly in exposed subjects than in controls. Our studies highlighted the association of epigenetic modifications of DNA with low benzene exposure.

  1. Alkyl- and fluoroalkyltrialkoxysilanes for wettability modification

    Energy Technology Data Exchange (ETDEWEB)

    Dopierala, Katarzyna, E-mail: katarzyna.dopierala@put.poznan.pl [Institute of Chemical Technology and Engineering, Poznan University of Technology, Pl. Marii Skłodowskiej-Curie 2, Poznań 60-965 (Poland); Maciejewski, Hieronim [Poznań Science and Technology Park, Rubież 46, Poznań 61-612 (Poland); Faculty of Chemistry, Adam Mickiewicz University, Grunwaldzka 6, Poznań 60-780 (Poland); Karasiewicz, Joanna [Poznań Science and Technology Park, Rubież 46, Poznań 61-612 (Poland); Prochaska, Krystyna [Institute of Chemical Technology and Engineering, Poznan University of Technology, Pl. Marii Skłodowskiej-Curie 2, Poznań 60-965 (Poland)

    2013-10-15

    Alkyl- and fluoroalkyltriethoxysilanes were investigated as potential surface modifiers. Many aspects concerning trialkoxysilanes were discussed, starting from hydrolysis of silanes in water solutions, the effect of this hydrolysis on the surface tension, wettability of the modified surface to the morphology of the modified surface. Surface tension and contact angle measurements as well as scanning electron microscopy were used to characterise alkyl- and fluoroalkyltriethoxysilanes and their ability to modify the wettability of glass. The effect of such modification was superhydrophobic surface with high values of contact angles. Superhydrophobic behaviour was observed as a result of two-step modification providing increased surface roughness thanks to the use of different size silica particles and surface chemical modification with fluorosilane molecules.

  2. PREVIEW Behavior Modification Intervention Toolbox (PREMIT)

    DEFF Research Database (Denmark)

    Kahlert, Daniela; Unyi-Reicherz, Annelie; Stratton, Gareth

    2016-01-01

    of behavior modification by providing a deeper understanding of successful intervention components. OBJECTIVE: To develop a physical activity and dietary behavior modification intervention toolbox (PREMIT) that fulfills current requirements of being theory-driven and evidence-based, comprehensively described......) implementing the intervention and assuring quality. RESULTS: PREMIT is based on a trans-theoretical approach referring to valid behavior modification theories, models and approaches. A major "product" of PREMIT is a matrix, constructed for use by onsite-instructors. The matrix includes objectives, tasks...... and feasible to evaluate. PREMIT is part of an intervention trial, which aims to prevent the onset of type-2 diabetes in pre-diabetics in eight clinical centers across the world by guiding them in changing their physical activity and dietary behavior through a group counseling approach. METHODS: The program...

  3. Compositional, physical and chemical modification of polylactide

    Directory of Open Access Journals (Sweden)

    M. Żenkiewicz

    2010-11-01

    Full Text Available Purpose: The purpose of this article was to review some of the modification methods applied to improve mechanical, barrier and/or surface properties of polylactide (PLA.Design/methodology/approach: The presented modification methods were classified into three groups due to the dominant role of compositional, physical or chemical factor effecting the most PLA properties.Findings: It was found that incorporation of small amounts of montmorillonite up to 5% leads to formation of a nanocomposite with enhanced tensile strength and improved barrier properties. Corona treatment of pure PLA and PLA contained MMT nanofiller causes a significant decrease in the water contact angle and does not essentially affect the diiodomethane contact angle. This treatment leads to an increase in surface free energy that is much more significant for pure PLA than for PLA containing MMT nanofiller. It was also found that with increasing number up to 1000 of laser pulses of energies 5 mJ/cm2 an increase in surface free energy was observed, while the next laser pulses caused decrease of this energy. The determination and comparison of the influence of 3 wt.% of trimethylopropane trimethacylate (TMPTA and 3 wt.% of trially isocyanurate (TAIC crosslinking agents on the thermomechanical properties of electron beam irradiated PLA was reported.Research limitations/implications: A number of various modification methods are widely reported in literature. In this article a review of only such modification methods is presented, which are in line with the newest trends in polymer industry and science.Practical implications: There are a number of PLA properties, which need to be improved to satisfy specific application conditions. For that reasons researches are leading to find suitable modification methods to improve selected properties of PLA.Originality/value: This article presents some of modification methods, which are in line with the newest trends in polymer industry and

  4. Development of an Improved Permeability Modification Simulator

    Energy Technology Data Exchange (ETDEWEB)

    Gao, H.W.; Elphnick, J.

    1999-03-09

    This report describes the development of an improved permeability modification simulator performed jointly by BDM Petroleum Technologies and Schlumberger Dowell under a cooperative research and development agreement (CRADA) with the US Department of Energy. The improved simulator was developed by modifying NIPER's PC-GEL permeability modification simulator to include a radial model, a thermal energy equation, a wellbore simulator, and a fully implicit time-stepping option. The temperature-dependent gelation kinetics of a delayed gel system (DGS) is also included in the simulator.

  5. Chromatin proteins and modifications as drug targets

    DEFF Research Database (Denmark)

    Helin, Kristian; Dhanak, Dashyant

    2013-01-01

    A plethora of groundbreaking studies have demonstrated the importance of chromatin-associated proteins and post-translational modifications of histones, proteins and DNA (so-called epigenetic modifications) for transcriptional control and normal development. Disruption of epigenetic control...... is a frequent event in disease, and the first epigenetic-based therapies for cancer treatment have been approved. A generation of new classes of potent and specific inhibitors for several chromatin-associated proteins have shown promise in preclinical trials. Although the biology of epigenetic regulation...

  6. Structural dynamic modification using additive damping

    Indian Academy of Sciences (India)

    B C Nakra

    2000-06-01

    In order to control dynamic response in structures and machines, modofications using additive viscoelastic damping materials are highlighted. The techniques described for analysis include analytical methods for structural elements, FEM and perturbation methods for reanalysis or structural dynamic modifications for complex structures. Optimisation techniques are used for damping effectiveness include multi-parameter optimisatoin techniques and a technique using dynamic sensitivity analysis and structural dynamic modification. These have been applied for optimum dynamic design of structures incorporating viscoelastic damping. Some current trends for vibraton control are also discussed.

  7. Stability of Strongly Gauduchon Manifolds under Modifications

    CERN Document Server

    Popovici, Dan

    2010-01-01

    In our previous works on deformation limits of projective and Moishezon manifolds, we introduced and made crucial use of the notion of strongly Gauduchon metrics as a reinforcement of the earlier notion of Gauduchon metrics. Using direct and inverse images of closed positive currents of type $(1, \\, 1)$ and regularisation, we now show that compact complex manifolds carrying strongly Gauduchon metrics are stable under modifications. This stability property, known to fail for compact K\\"ahler manifolds, mirrors the modification stability of balanced manifolds proved by Alessandrini and Bassanelli.

  8. Epigenomic Modification and Epigenetic Regulation in Rice

    Institute of Scientific and Technical Information of China (English)

    Yu Zhao; Dao-Xiu Zhou

    2012-01-01

    Epigenomes including genome-wide histone modification and DNA methylation profiles are important for genome activity and for defining gene expression patterns of plant development and responses to various environmental conditions.Rice is the most important crop plant and serves as a model for cereal genomics.Rice epigenomic landscape is emerging and the function of chromatin modification regulators in gene expression,transposon repression and plant development is being characterized.Epigenomic variation that gives rise to stable or transgenerational heritable epialleles related to variation of important agronomical traits or stress responses is being characterized in rice.Implication of epigenomic variation in rice heterosis is being exploited.

  9. Surface modification to prevent oxide scale spallation

    Science.gov (United States)

    Stephens, Elizabeth V; Sun, Xin; Liu, Wenning; Stevenson, Jeffry W; Surdoval, Wayne; Khaleel, Mohammad A

    2013-07-16

    A surface modification to prevent oxide scale spallation is disclosed. The surface modification includes a ferritic stainless steel substrate having a modified surface. A cross-section of the modified surface exhibits a periodic morphology. The periodic morphology does not exceed a critical buckling length, which is equivalent to the length of a wave attribute observed in the cross section periodic morphology. The modified surface can be created using at least one of the following processes: shot peening, surface blasting and surface grinding. A coating can be applied to the modified surface.

  10. Beam Distribution Modification By Alfven Modes

    Energy Technology Data Exchange (ETDEWEB)

    White, R. B.; Gorelenkov, N.; Heidbrink, W. W.; Van Zeeland, M. A.

    2010-01-25

    Modification of a deuterium beam distribution in the presence of low amplitude Toroidal Alfven (TAE) eigenmodes and Reversed Shear Alfven (RSAE) eigenmodes in a toroidal magnetic confinement device is examined. Comparison with experimental data shows that multiple low amplitude modes can account for significant modification of high energy beam particle distributions. It is found that there is a stochastic threshold for beam transport, and that the experimental amplitudes are only slightly above this threshold. The modes produce a substantial central flattening of the beam distribution.

  11. Beam Distribution Modification by Alfven Modes

    Energy Technology Data Exchange (ETDEWEB)

    White, R. B.; Gorelenkov, N.; Heidbrink, W. W.; Van Zeeland, M. A.

    2010-04-03

    Modification of a deuterium beam distribution in the presence of low amplitude Toroidal Alfven (TAE) eigenmodes and Reversed Shear Alfven (RSAE) eigenmodes in a toroidal magnetic confinement device is examined. Comparison with experimental data shows that multiple low amplitude modes can account for significant modification of high energy beam particle distributions. It is found that there is a stochastic threshold for beam transport, and that the experimental amplitudes are only slightly above this threshold. The modes produce a substantial central flattening of the beam distribution.

  12. Epigenetic Modifications in Pediatric Acute Lymphoblastic Leukemia

    Directory of Open Access Journals (Sweden)

    Michael James Burke

    2014-05-01

    Full Text Available Aberrant epigenetic modifications are well-recognized drivers for oncogenesis. Pediatric acute lymphoblastic leukemia (ALL is no exception and serves as a model toward the significant impact these heritable alterations can have in leukemogenesis. In this brief review, we will focus on the main aspects of epigenetics which control leukemogenesis in pediatric ALL, mainly DNA methylation, histone modification and microRNA alterations. As we continue to gain better understanding of the driving mechanisms for pediatric ALL at both diagnosis and relapse, therapeutic interventions directed toward these pathways and mechanisms can be harnessed and introduced into clinical trials for pediatric ALL.

  13. 42 CFR 84.35 - Changes or modifications of approved respirators; issuance of modification of certificate of...

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 1 2010-10-01 2010-10-01 false Changes or modifications of approved respirators... modifications of approved respirators; issuance of modification of certificate of approval. (a) Each applicant may, if he desires to change any feature of an approved respirator, request a modification of...

  14. Covalent Modification of DNA Regulates Memory Formation

    National Research Council Canada - National Science Library

    Miller, Courtney A; Sweatt, J. David

    2007-01-01

    ... during mitosis. The mechanisms that accomplish this are a set of posttranslational modifications of DNA and chromatin that alter gene expression patterns. Rapidly accumulating evidence suggests that the nervous system has co-opted these epigenetic mechanisms utilized during development for the generation of long-term behavioral memories in adulthood ( S...

  15. Extreme exercise and oxidative DNA modification

    DEFF Research Database (Denmark)

    Poulsen, H E; Loft, S; Vistisen, K

    1996-01-01

    increased the rate of oxidative DNA modification by 33% (95% confidence limits, 3-67%; P DNA repair, oxidation of the nucleotide pool from mitochondrial...... DNA and/or from cell turnover. Oxidative stress to DNA points to a risk for the development of cancer and premature ageing from extreme exercise....

  16. In-Medium Modifications of Hadron Properties

    CERN Document Server

    Tawfik, A

    2006-01-01

    The in-medium modifications of hadron properties are briefly discussed. We restrict the discussion to the lattice QCD calculations for the hadron masses, screening masses, decay constants and wave functions. We review the progress made so far and describe how to broaden its horizon.

  17. Some peculiarities of bitumen emulsion modification

    Directory of Open Access Journals (Sweden)

    A. Batyrbayev

    2013-09-01

    Full Text Available Polymer modification of bitumen emulsions obtained from bitumen of domestic production with the use of several commercial emulsifiers was studied. The influence of the polymer modifier concentration on bitumen emulsion physical-mechanical properties was considered. Possibility of obtaining of modified bitumen emulsion with high impact resistance was shown.

  18. Microscale surface modifications for heat transfer enhancement.

    Science.gov (United States)

    Bostanci, Huseyin; Singh, Virendra; Kizito, John P; Rini, Daniel P; Seal, Sudipta; Chow, Louis C

    2013-10-09

    In this experimental study, two surface modification techniques were investigated for their effect on heat transfer enhancement. One of the methods employed the particle (grit) blasting to create microscale indentations, while the other used plasma spray coating to create microscale protrusions on Al 6061 (aluminum alloy 6061) samples. The test surfaces were characterized using scanning electron microscopy (SEM) and confocal scanning laser microscopy. Because of the surface modifications, the actual surface area was increased up to 2.8× compared to the projected base area, and the arithmetic mean roughness value (Ra) was determined to vary from 0.3 μm for the reference smooth surface to 19.5 μm for the modified surfaces. Selected samples with modified surfaces along with the reference smooth surface were then evaluated for their heat transfer performance in spray cooling tests. The cooling system had vapor-atomizing nozzles and used anhydrous ammonia as the coolant in order to achieve heat fluxes up to 500 W/cm(2) representing a thermal management setting for high power systems. Experimental results showed that the microscale surface modifications enhanced heat transfer coefficients up to 76% at 500 W/cm(2) compared to the smooth surface and demonstrated the benefits of these practical surface modification techniques to enhance two-phase heat transfer process.

  19. Starch--value addition by modification.

    Science.gov (United States)

    Tharanathan, Rudrapatnam N

    2005-01-01

    Starch is one of the most important but flexible food ingredients possessing value added attributes for innumerable industrial applications. Its various chemically modified derivatives offer a great scope of high technological value in both food and non-food industries. Modified starches are designed to overcome one or more of the shortcomings, such as loss of viscosity and thickening power upon cooking and storage, particularly at low pH, retrogradation characteristics, syneresis, etc., of native starches. Oxidation, esterification, hydroxyalkylation, dextrinization, and cross-linking are some of the modifications commonly employed to prepare starch derivatives. In a way, starch modification provides desirable functional attributes as well as offering economic alternative to other hydrocolloid ingredients, such as gums and mucilages, which are unreliable in quality and availability. Resistant starch, a highly retrograded starch fractionformed upon food processing, is another useful starch derivative. It exhibits the beneficial physiological effects of therapeutic and nutritional values akin to dietary fiber. There awaits considerable opportunity for future developments, especially for tailor-made starch derivatives with multiple modifications and with the desired functional and nutritional properties, although the problem of obtaining legislative approval for the use of novel starch derivatives in processed food formulations is still under debate. Nevertheless, it can be predicted that new ventures in starch modifications and their diverse applications will continue to be of great interest in applied research.

  20. Genetic modifications for personal enhancement: a defence.

    Science.gov (United States)

    Murphy, Timothy F

    2014-04-01

    Bioconservative commentators argue that parents should not take steps to modify the genetics of their children even in the name of enhancement because of the damage they predict for values, identities and relationships. Some commentators have even said that adults should not modify themselves through genetic interventions. One commentator worries that genetic modifications chosen by adults for themselves will undermine moral agency, lead to less valuable experiences and fracture people's sense of self. These worries are not justified, however, since the effects of modification will not undo moral agency as such. Adults can still have valuable experiences, even if some prior choices no longer seem meaningful. Changes at the genetic level will not always, either, alienate people from their own sense of self. On the contrary, genetic modifications can help amplify choice, enrich lives and consolidate identities. Ultimately, there is no moral requirement that people value their contingent genetic endowment to the exclusion of changes important to them in their future genetic identities. Through weighing risks and benefits, adults also have the power to consent to, and assume the risks of, genetic modifications for themselves in a way not possible in prenatal genetic interventions.

  1. Alignment modification for pencil eye shields

    Energy Technology Data Exchange (ETDEWEB)

    Evans, M.D.; Pla, M.; Podgorsak, E.B. (McGill Univ., Quebec (Canada))

    1989-01-01

    Accurate alignment of pencil beam eye shields to protect the lens of the eye may be made easier by means of a simple modification of existing apparatus. This involves drilling a small hole through the center of the shield to isolate the rayline directed to the lens and fabricating a suitable plug for this hole.

  2. Modification of Behavior: Application in Classroom Management.

    Science.gov (United States)

    Ille, Sister Janiece; Bezzi, D. R.

    The emphasis of this research report was on modification of classroom behavior. After a brief introduction, the two main learning theories of Gestalt-field psychology and stimulus-response association were investigated. The importance of the individual in modifying his own behavior was stressed in Gestalt-field psychology. The importance of…

  3. 7 CFR 987.35 - Modifications.

    Science.gov (United States)

    2010-01-01

    ... RIVERSIDE COUNTY, CALIFORNIA Order Regulating Handling Marketing Policy § 987.35 Modifications. In the event the Committee subsequently determines that the marketing policy should be modified due to changing... Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing...

  4. A Suggested Modification to Maslow's Need Hierarchy

    Science.gov (United States)

    Groves, David L.; And Others

    1975-01-01

    Since its development, Maslow's need hierarchy has been criticized and applauded. This investigation was undertaken to explore a modification of the upper levels of the need hierarchy based upon the application of power, competition, and achievement to self, as well as the concept of "other directed." (Author)

  5. 75 FR 41530 - Petitions for Modification; Correction

    Science.gov (United States)

    2010-07-16

    ... Safety and Health Administration Petitions for Modification; Correction AGENCY: Mine Safety and Health Administration, Labor. ACTION: Notice; correction. SUMMARY: The Mine Safety and Health Administration (MSHA... Affected: 30 CFR 75.507-1(a) (Electric equipment other than power-connection points; outby the last...

  6. A Suggested Modification to Maslow's Need Hierarchy

    Science.gov (United States)

    Groves, David L.; And Others

    1975-01-01

    Since its development, Maslow's need hierarchy has been criticized and applauded. This investigation was undertaken to explore a modification of the upper levels of the need hierarchy based upon the application of power, competition, and achievement to self, as well as the concept of "other directed." (Author)

  7. A Classroom Modification of the Ames Test.

    Science.gov (United States)

    Yavornitzky, Joseph; Trzeciak, Victor

    1979-01-01

    A modification of the Ames test for detecting carcinogens and mutagens using a strain of bacteria is described. A suggestion is given for checking the correctness of procedures by using particular hair dyes which have been shown to be mutogenic. (Author/SA)

  8. An Energy Absorber with Force Modificator

    Institute of Scientific and Technical Information of China (English)

    SU Hao; ZHANG Xiaowei; YU Tongxi

    2006-01-01

    Thin-walled tubes are extensively applied in engineering,especially in vehicle structures to resist axial or traversal impact loads,for their excellent energy absorbing capacity.However,in the axial deformation mode,the force history has an extremely high peak force which may bring not only fatal injury to occupants but also damage to structures,cargo and environment.Aiming to develop energy absorbers with impact-force modificator,square metal tube with force modificator is investigated which can monitor the force-deformation history of the tube.A small device is designed to serve as an impact-force modificator,which introduces desired imperfections to the square tube just before the impact happens between the impactor and the tube,so as to reduce the peak force.Prototypes with various governing parameters were manufactured and tested both quasi-statically and dynamically to study the effects of these parameters on the characteristics of energy absorption.The results show that the force modificator can achieve the desired reduction of the peak force well whilst remaining the specific energy absorption capacity of the original square tube.With future improvements,it could be applied to vehicles or roadside safety hardware to mitigate the consequences produced by traffic accidents.

  9. 75 FR 27002 - Petitions for Modification

    Science.gov (United States)

    2010-05-13

    ... bags, rock dust, mine gob, earth--rock and dirt, gravel, refuse, or other similar material that is...) . Modification Request: The petitioner proposes to use Model J251-D/E Face Drill, manufactured by J.H. Fletcher & Company. The petitioner states that: (1) The J251-D/E Face Drill consists of an electric drill...

  10. SUMO 2016 – Traffic, Mobility, and Logistics

    OpenAIRE

    Blokpoel, Robbin; Joueiai, Mahtab; Barthauer, Mirko; Friedrich, Bernhard; Richter, Andreas; Friedl, Hartmut; Scholz, Michael; Krumnow, Mario; Flötteröd, Yun-Pang; Erdmann, Jakob; Busquets, Dídac; Francis, Raymond; Tsotskas, Christos; North, Robin; Galatioto, Fabio

    2016-01-01

    Dear reader, You are holding in your hands a volume of the series „Reports of the DLR-Institute of Transportation Systems“. We are publishing in this series fascinating, scientific topics from the Institute of Trans- portation Systems of the German Aerospace Center (Deutsches Zentrum für Luft- und Raumfahrt e.V. – DLR) and from his environment. We are providing libraries with a part of the circulation. Outstanding scientific contributions and dissertations are here published as...

  11. Determinants of Treatment Modification in Hypercholesterolemic Patients

    Science.gov (United States)

    Wang, Ko-Fan; Wu, Cheng-Hsueh; Chang, Chun-Chin; Chen, Lung-Ching; Wang, Kang-Ling; Lu, Tse-Min; Lin, Shing-Jong; Chiang, Chern-En

    2017-01-01

    Background There is a lack of knowledge of those contemporary factors associated with modifying subtherapeutic treatments in hypercholesterolemic patients. The aim of this study was to assess determinants of treatment modification in patients not attaining their low-density lipoprotein cholesterol goals. Methods The centralized Pan-Asian survey on the under-treatment of hypercholesterolemia enrolled patients taking stable lipid-lowering medications. The study physicians then determined existing patient treatments, which were to be continued or modified when treatments failed. The patient questionnaire surveying patient attitudes and perceptions toward their hypercholesterolemia management was prospectively collected. The odds ratios (ORs) (95% confidence intervals) were calculated. Results Among the 420 patients included for analysis, 35.7% were designated for planned treatment modification. Those patients assigned to treatment modification were more likely to have a family history of premature coronary heart disease (40% vs. 19%), an indication for secondary prevention (76% vs. 61%), elevated triglyceride (60% vs. 48%) and fasting sugar (84% vs. 67%), and were less adherent to their medications (29% vs. 12%) than patients assigned to treatment continuation. Patient recognition of treatment failure [OR, 1.82 (1.13-2.94)], the lower frequency of cholesterol checkup [OR, 2.40 (1.41-4.08)], patient satisfaction with provided cholesterol information [OR, 2.30 (1.21-4.39)], and their feelings toward cholesterol management [OR, 0.25 (0.10-0.62) and 3.80 (2.28-6.32)] for confusion and no strong feeling, respectively were determinants of the treatment modification assignment. Conclusions There was a large gap between evidence-based goals and modification of subtherapeutic treatments, particularly among patients with lower treatment satisfaction and better compliance. Our findings have emphasized the need to further reduce inertia in implementing hypercholesterolemia

  12. Improved Collision Search for Hash Functions: New Advanced Message Modification

    Science.gov (United States)

    Naito, Yusuke; Ohta, Kazuo; Kunihiro, Noboru

    In this paper, we discuss the collision search for hash functions, mainly in terms of their advanced message modification. The advanced message modification is a collision search tool based on Wang et al.'s attacks. Two advanced message modifications have previously been proposed: cancel modification for MD4 and MD5, and propagation modification for SHA-0. In this paper, we propose a new concept of advanced message modification, submarine modification. As a concrete example combining the ideas underlying these modifications, we apply submarine modification to the collision search for SHA-0. As a result, we show that this can reduce the collision search attack complexity from 239 to 236 SHA-0 compression operations.

  13. 77 FR 37926 - Petitions for Modification of Application of Existing Mandatory Safety Standards

    Science.gov (United States)

    2012-06-25

    ... equipment and conductors; permissibility). Modification Request: The petitioner requests a modification of... CFR 75.1002(a) (Installation of electric equipment and conductors; permissibility). Modification... equipment and conductors; permissibility). Modification Request: The petitioner requests a modification...

  14. Nuclear modifications of Parton Distribution Functions

    Science.gov (United States)

    Adeluyi, Adeola Adeleke

    This dissertation addresses a central question of modern nuclear physics: how does the behavior of fundamental degrees of freedom (quarks and gluons) change in the nuclear environment? This is an important aspect of experimental studies at current facilities such as the Relativistic Heavy Ion Collider (RHIC) at Brookhaven National Laboratory and the Continuous Electron Beam Accelerator Facility (CEBAF) at the Thomas Jefferson National Laboratory (JLAB). It is also highly relevant to planned experimental efforts at the Large Hadron Collider (LHC) and the future Electron Ion Collider (EIC). All these facilities probe matter via collisions involving nuclei; thus complications arise due to the presence of the attendant nuclear medium. Theoretical efforts to understand and interpret experimental results from such collisions are therefore largely dependent on the resolution of this question. The development of nuclear physics demonstrates that theoretical description is most efficient in terms of the effective degrees of freedom relevant to the scale (energy) being probed. Thus at low energies, nuclei are described as bound states of protons and neutrons (nucleons). At higher energies, the nucleons are no longer elementary, but are revealed to possess an underlying substructure: they are made up of quarks and gluons, collectively termed partons. The mometum distributions of these partons in the nucleon are referred to as Parton Distribution Functions (PDFs). Parton distributions can be determined from experimental measurements of structure functions. The ratio of nuclear structure functions to nucleon structure functions (generically referred to as nuclear ratio) is a measure of the nuclear modifications of the free nucleon PDFs. Thus a study of the nuclear ratio suffices to gain an understanding of nuclear modifications. In this dissertation we aim to describe theoretically nuclear modifications in a restricted region where the nuclear ratio is less than unity, the so

  15. 30 CFR 18.81 - Field modification of approved (permissible) equipment; application for approval of modification...

    Science.gov (United States)

    2010-07-01

    ... TESTING, EVALUATION, AND APPROVAL OF MINING PRODUCTS ELECTRIC MOTOR-DRIVEN MINE EQUIPMENT AND ACCESSORIES Machines Assembled With Certified or Explosion-Proof Components, Field Modifications of Approved Machines... and Certification Center, 765 Technology Drive, Triadelphia, WV 26059. (b) Proposed...

  16. Graphite Surface Modification by Heterogeneous Nucleation Process

    Institute of Scientific and Technical Information of China (English)

    CAO Ran; LI Hongxia

    2006-01-01

    Flaky graphite particles were coated by ZrOCl2·8H2O as precursors by heterogeneous nucleation process.The effects of factors such as pH values (2.4-5.1),concentration of the precursor solution (0.005-0.1 mol·L-1 ) , mixing method of graphite and precursor solution on the surface modification of graphite were studied. Result shows that: 1) the preferable technical process for heterogeneous nucleation modified graphite is to mix the graphite suspension and precursor solution with concentration 0. 025 mol·L -1 and then drip ammonia water to adjust the pH value to 3.6; 2)By surface modification, the ZrO2 particles are evenly coated on graphite surface and therefore improve oxidation resistance and dispersion ability of graphite.

  17. SKELETAL MODIFICATIONS IN TURNER AND KLINEFELTER SYNDROMES

    Directory of Open Access Journals (Sweden)

    Adriana Balan

    2011-12-01

    Full Text Available The study aims at identifying the growth disorders in children and adolescents suffering from Turner (45X0 and Klinefelter syndromes (47XXY, in order to correctly establish subsequent treatment strategies. To this extent, a group of children and adolescents formed of 18 girls (9.5-17.5 year-old and 8 boys (14.5-18.5 year-old was investigated. Analysis of these subjects demonstrated dimensional modifications of the basis of the skull, of the maxillary and mandible length. The absence of a chromsome X or its supranumerary presence influences especially the shape and size of the skull basis, of the mandible and of the intermaxillary relations. Chromosomial X aneuploidy determines dimensional modifications of the anterior and posterior facial height.

  18. Video Playback Modifications for a DSpace Repository

    Directory of Open Access Journals (Sweden)

    Keith Gilbertson

    2016-01-01

    Full Text Available This paper focuses on modifications to an institutional repository system using the open source DSpace software to support playback of digital videos embedded within item pages. The changes were made in response to the formation and quick startup of an event capture group within the library that was charged with creating and editing video recordings of library events and speakers. This paper specifically discusses the selection of video formats, changes to the visual theme of the repository to allow embedded playback and captioning support, and modifications and bug fixes to the file downloading subsystem to enable skip-ahead playback of videos via byte-range requests. This paper also describes workflows for transcoding videos in the required formats, creating captions, and depositing videos into the repository.

  19. Chitosan Modification and Pharmaceutical/Biomedical Applications

    Directory of Open Access Journals (Sweden)

    Jiali Zhang

    2010-06-01

    Full Text Available Chitosan has received much attention as a functional biopolymer for diverse applications, especially in pharmaceutics and medicine. Our recent efforts focused on the chemical and biological modification of chitosan in order to increase its solubility in aqueous solutions and absorbability in the in vivo system, thus for a better use of chitosan. This review summarizes chitosan modification and its pharmaceutical/biomedical applications based on our achievements as well as the domestic and overseas developments: (1 enzymatic preparation of low molecular weight chitosans/chitooligosaccharides with their hypocholesterolemic and immuno-modulating effects; (2 the effects of chitin, chitosan and their derivatives on blood hemostasis; and (3 synthesis of a non-toxic ion ligand—D-Glucosaminic acid from Oxidation of D-Glucosamine for cancer and diabetes therapy.

  20. Fault diagnosis based on controller modification

    DEFF Research Database (Denmark)

    Niemann, Hans Henrik

    2015-01-01

    Detection and isolation of parametric faults in closed-loop systems will be considered in this paper. A major problem is that a feedback controller will in general reduce the effects from variations in the systems including parametric faults on the controlled output from the system. Parametric...... time. A negative effect of increasing the amplitude of the auxiliary input is that the disturbances in the external output will be increased and consequently reduce the closed-loop performance. This problem can be handled by using a modification of the feedback controller. Applying the YJBK......-parameterization (after Youla, Jabr, Bongiorno and Kucera) for the controller, it is possible to modify the feedback controller with a minor effect on the closed-loop performance in the fault-free case and at the same time optimize the detection and isolation in a faulty case. Controller modification in connection...

  1. Chitosan Modification and Pharmaceutical/Biomedical Applications

    Science.gov (United States)

    Zhang, Jiali; Xia, Wenshui; Liu, Ping; Cheng, Qinyuan; Tahirou, Talba; Gu, Wenxiu; Li, Bo

    2010-01-01

    Chitosan has received much attention as a functional biopolymer for diverse applications, especially in pharmaceutics and medicine. Our recent efforts focused on the chemical and biological modification of chitosan in order to increase its solubility in aqueous solutions and absorbability in the in vivo system, thus for a better use of chitosan. This review summarizes chitosan modification and its pharmaceutical/biomedical applications based on our achievements as well as the domestic and overseas developments: (1) enzymatic preparation of low molecular weight chitosans/chitooligosaccharides with their hypocholesterolemic and immuno-modulating effects; (2) the effects of chitin, chitosan and their derivatives on blood hemostasis; and (3) synthesis of a non-toxic ion ligand—D-Glucosaminic acid from Oxidation of D-Glucosamine for cancer and diabetes therapy. PMID:20714418

  2. Computational Methods for Modification of Metabolic Networks

    Directory of Open Access Journals (Sweden)

    Takeyuki Tamura

    2015-01-01

    Full Text Available In metabolic engineering, modification of metabolic networks is an important biotechnology and a challenging computational task. In the metabolic network modification, we should modify metabolic networks by newly adding enzymes or/and knocking-out genes to maximize the biomass production with minimum side-effect. In this mini-review, we briefly review constraint-based formalizations for Minimum Reaction Cut (MRC problem where the minimum set of reactions is deleted so that the target compound becomes non-producible from the view point of the flux balance analysis (FBA, elementary mode (EM, and Boolean models. Minimum Reaction Insertion (MRI problem where the minimum set of reactions is added so that the target compound newly becomes producible is also explained with a similar formalization approach. The relation between the accuracy of the models and the risk of overfitting is also discussed.

  3. Diagonal chromatography to study plant protein modifications.

    Science.gov (United States)

    Walton, Alan; Tsiatsiani, Liana; Jacques, Silke; Stes, Elisabeth; Messens, Joris; Van Breusegem, Frank; Goormachtig, Sofie; Gevaert, Kris

    2016-08-01

    An interesting asset of diagonal chromatography, which we have introduced for contemporary proteome research, is its high versatility concerning proteomic applications. Indeed, the peptide modification or sorting step that is required between consecutive peptide separations can easily be altered and thereby allows for the enrichment of specific, though different types of peptides. Here, we focus on the application of diagonal chromatography for the study of modifications of plant proteins. In particular, we show how diagonal chromatography allows for studying proteins processed by proteases, protein ubiquitination, and the oxidation of protein-bound methionines. We discuss the actual sorting steps needed for each of these applications and the obtained results. This article is part of a Special Issue entitled: Plant Proteomics--a bridge between fundamental processes and crop production, edited by Dr. Hans-Peter Mock.

  4. Restriction/modification polypeptides, polynucleotides, and methods

    Energy Technology Data Exchange (ETDEWEB)

    Westpheling, Janet; Chung, DaeHwan; Huddleston, Jennifer; Farkas, Joel A

    2015-02-24

    The present invention relates to the discovery of a novel restriction/modification system in Caldicellulosiruptor bescii. The discovered restriction enzyme is a HaeIII-like restriction enzyme that possesses a thermophilic activity profile. The restriction/modification system also includes a methyltransferase, M.CbeI, that methylates at least one cytosine residue in the CbeI recognition sequence to m.sup.4C. Thus, the invention provides, in various aspects, isolated CbeI or M.CbeI polypeptides, or biologically active fragments thereof; isolated polynucleotides that encode the CbeI or M.CbeI polypeptides or biologically active fragments thereof, including expression vectors that include such polynucleotide sequences; methods of digesting DNA using a CbeI polypeptide; methods of treating a DNA molecule using a M.CbeI polypeptide; and methods of transforming a Caldicellulosiruptor cell.

  5. CTL Model Update for System Modifications

    CERN Document Server

    Ding, Yulin; Zhang, Yan; Zhang, Y; 10.1613/jair.2420

    2011-01-01

    Model checking is a promising technology, which has been applied for verification of many hardware and software systems. In this paper, we introduce the concept of model update towards the development of an automatic system modification tool that extends model checking functions. We define primitive update operations on the models of Computation Tree Logic (CTL) and formalize the principle of minimal change for CTL model update. These primitive update operations, together with the underlying minimal change principle, serve as the foundation for CTL model update. Essential semantic and computational characterizations are provided for our CTL model update approach. We then describe a formal algorithm that implements this approach. We also illustrate two case studies of CTL model updates for the well-known microwave oven example and the Andrew File System 1, from which we further propose a method to optimize the update results in complex system modifications.

  6. Amalyse modifications during production and application

    DEFF Research Database (Denmark)

    Nielsen, Connie

    . The commercial a-amylases are produced by fermentations of genetically modified bacteria, where the native gene has been manipulated to code for an enzyme with improved performance characteristics. Despite the genome only contains copies of the same a-amylase gene, multiple different a-amylase forms varying in p......I value can be identified when analysing the final enzyme product, hereby indicating that the protein is modified. The purpose of the present Ph.D. project is to identify differences between the multiple forms of a-amylase derived from the same gene and determine if or how these various modifications...... affect the activity and wash performance. The focus of the present project is on a-amylases fromBacillus species used in the detergent industry. The thesis begins with an introduction to Bacillus a-amylases, modifications which could cause the multiple forms varying in pI value and mass spectrometry (MS...

  7. Nursing researchers' modifications of Ricoeur's hermeneutic phenomenology.

    Science.gov (United States)

    Singsuriya, Pagorn

    2015-12-01

    Paul Ricoeur's hermeneutic phenomenology has proved to be very helpful in guiding nursing researchers' qualitative analysis of interview transcripts. Modifying Ricoeur's philosophy, a number of nursing researchers have developed their own interpretive methods and shared them, along with their experience, with research community. Major contributors who published papers directly presenting their modifications of Ricoeur's theory include Rene Geanellos (2000), Lena Wiklund, Lisbet Lindholm and Unni Å. Lindström (2002), Anders Lindseth and Astrid Norberg (2004) and Pia Sander Dreyer and Birthe D Pedersen (2009). The aim of this article was to delineate differences among these methods. Descriptive presentation of each method side by side makes clear the differences among them. In addition, Ricoeur's hermeneutic theory is portrayed and compared with the modifications. It is believed that differences that are found can stimulate further thoughts on how to apply Ricoeur's theory in qualitative research in nursing.

  8. Behavioral modification in choice process of Drosophila

    Institute of Scientific and Technical Information of China (English)

    WANG; Shunpeng; (王顺鹏); TANG; Shiming; (唐世明); LI; Yan; (李; 岩); GUO; Aike; (郭爱克)

    2003-01-01

    In visual operant conditioning of Drosophila at the flight simulator, only motor output of flies--yaw torque--is recorded, which is involved in the conditioning process. The current study used a newly-designed data analysis method to study the torque distribution of Drosophila. Modification of torque distribution represents the effects of operant conditioning on flies' behavioral mode. Earlier works[10] showed that, when facing contradictory visual cues, flies could make choices based upon the relative weightiness of different cues, and it was demonstrated that mushroom bodies might play an important role in such choice behavior. The new "torque-position map" method was used to explore the CS-US associative learning and choice behavior in Drosophila from the aspect of its behavioral mode. Finally, this work also discussed various possible neural bases involved in visual associative learning, choice processing and modification processing of the behavioral mode in the visual operant conditioning of Drosophila.

  9. Mechanism of Surface Modification for Sericite

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Surface modification of sericite by wet method was conducted with the addition of 1.0% (w/w) silane. The resulting wetting contact angle and activity ratio of sericite were 130° and 98% respectively.Good pre-evaluation indexes of oil value (40.8%) and dispersivity (14.0 mL) were obtained. When 30% of sericite was filled into acrylonitrile butadiene styrene(ABS) plastic, the bending strength and tensile strength of the composite material were reduced by 7% and 14.3% in comparison to those of pure ABS plastic, while the rigidity was increased by 3 times, and the impact strength and breaking elongation were reduced significantly.The mechanism of surface modification was investigated and the configuration of silane coupling agent on the surface of sericite was given. Infrared (IR) spectroscopic analysis indicates that the adsorption of silane on the surface of sericite belongs to chemical adsorption.

  10. Quantitative proteomic approaches to studying histone modifications.

    Science.gov (United States)

    Zee, Barry M; Young, Nicolas L; Garcia, Benjamin A

    2011-01-01

    Histone post-translational modifications (PTMs) positively and negatively regulate gene expression, and are consequently a vital influence on the genomic profile of all eukaryotic species. The study of histone PTMs using classical methods in molecular biology, such as immunofluorescence and Western blotting, is challenging given the technical issues of the approaches, and chemical diversity and combinatorial patterns of the modifications. In light of these many technical limitations, mass spectrometry (MS) is emerging as the most unbiased and rigorous experimental platform to identify and quantify histone PTMs in a high-throughput manner. This review covers the latest developments in mass spectrometry for the analysis of histone PTMs, with the hope of inspiring the continued integration of proteomic, genomic and epigenetic research.

  11. Optical Marking of Alcohol Induced Hemoglobin Modification

    CERN Document Server

    Vlokh, R; Moroz, O; Nastishin, Yu; Dudok, K; Dudok, T; Grinchishin, N; Nechiporenko, I; Hul, A

    2007-01-01

    It has been shown that conformational modifications of Hb induced by ethanol consumption can be visualized in optical spectra studying oxygenation kinetics of hemoglobin or mixing hemoglobin with Cibacron blue dye. Better dye affinity of blood proteins extracted from alcoholised rats with respect to those from non-alcoholised ones confirms that ethanol and its metabolites induce structural pathologies in blood protein molecules. The detected changes for the case of the posterity of intoxicated animals may be explained as a post-translation modification, as well as a disturbance of the structure and function of tissue cellular gene mechanism for the blood creation. It is established that alcohol intake during first four months leads to the decrease of fractional weight of oxyhemoglobin and to the increase of methemoglobin amount in blood. Further alcohol consumption is accompanied by recovering of the normal level of hemoglobin derivatives in blood. Normalization of the fractional weight of hemoglobin derivati...

  12. PREVIEW Behavior Modification Intervention Toolbox (PREMIT)

    DEFF Research Database (Denmark)

    Kahlert, Daniela; Unyi-Reicherz, Annelie; Stratton, Gareth;

    2016-01-01

    of behavior modification by providing a deeper understanding of successful intervention components. OBJECTIVE: To develop a physical activity and dietary behavior modification intervention toolbox (PREMIT) that fulfills current requirements of being theory-driven and evidence-based, comprehensively described...... and feasible to evaluate. PREMIT is part of an intervention trial, which aims to prevent the onset of type-2 diabetes in pre-diabetics in eight clinical centers across the world by guiding them in changing their physical activity and dietary behavior through a group counseling approach. METHODS: The program...... development took five progressive steps, in line with the Public Health Action Cycle: (1) Summing-up the intervention goal(s), target group and the setting, (2) uncovering the generative psychological mechanisms, (3) identifying behavior change techniques and tools, (4) preparing for evaluation and (5...

  13. Design quality assurance for plant modifications

    Energy Technology Data Exchange (ETDEWEB)

    Ankrum, G.T.; Norkin, D.P.; Architzel, R.

    1985-11-01

    The US Nuclear Regulatory Commission's Office of Inspection and Enforcement has conducted direct inspections of the technical aspects of the nuclear plant design process for several plants under construction. It has also evaluated independent design verification programs and engineering assurance programs at a number of other plants under construction. Many of the lessons learned from these construction phase efforts are directly applicable to operating plants undergoing major modifications.

  14. Modification Semantics in Now-Relative Databases

    DEFF Research Database (Denmark)

    Torp, Kristian; Jensen, Christian Søndergaard; Snodgrass, R. T.

    2004-01-01

    , and with values of the type NOW + D, where D is a non-variable time duration. To accommodate these semantics, three new timestamp values are introduced. Finally, implementation is explored. We show how to represent the variable NOW with columns of standard SQL data types and give a mapping from SQL on NOW-relative......  data to standard SQL on these columns. The paper thereby completes the semantics, the querying, and the modification of now-relative databases.   Udgivelsesdato: DEC...

  15. Modification of Phenolic Oximes for Copper Extraction

    OpenAIRE

    Forgan, Ross Stewart

    2008-01-01

    The thesis deals with the modification of salicylaldoxime-based reagents used in hydrometallurgical extraction, addressing rational ligand design to tune copper(II) extractant strengths and also the development of reagents which are capable of transporting transition metal salts. Chapter 1 reviews current solvent extractant technology for metal recovery, including the limited knowledge of the effect of substituents on extractive efficacy. Advances in leaching technology have...

  16. Carbon Surface Modification for Enhanced Corrosion Resistance

    Science.gov (United States)

    2008-01-01

    2 R. Rayne,1 and R.A. Bayles1 1Chemistry Division 2SAIC Introduction: Case hardening by carburization has long been recognized to produce wear... carburization technique has been developed for intro- ducing carbon into stainless steel surfaces without formation of carbides.1,2 This surface modification...Michal, F. Ernst, H. Kahn, Y. Cao, F. Oba, N. Agarwal, and A.H. Heuer, “Carbon Supersaturation due to Paraequilibrium Carburization : Stainless

  17. SKELETAL MODIFICATIONS IN TURNER AND KLINEFELTER SYNDROMES

    OpenAIRE

    2011-01-01

    The study aims at identifying the growth disorders in children and adolescents suffering from Turner (45X0) and Klinefelter syndromes (47XXY), in order to correctly establish subsequent treatment strategies. To this extent, a group of children and adolescents formed of 18 girls (9.5-17.5 year-old) and 8 boys (14.5-18.5 year-old) was investigated. Analysis of these subjects demonstrated dimensional modifications of the basis of the skull, of the maxillary and mandibl...

  18. Firmware Modification Analysis in Programmable Logic Controllers

    Science.gov (United States)

    2014-03-27

    FORCE AIR UNIVERSITY AIR FORCE INSTITUTE OF TECHNOLOGY Wright-Patterson Air Force Base, Ohio DISTRIBUTION STATEMENT A: APPROVED FOR PUBLIC RELEASE...Faculty Department of Electrical and Computer Engineering Graduate School of Engineering and Management Air Force Institute of Technology Air University...controller ( PLC ). The Stuxnet worm, which became increasingly apparent in the summer of 2010, has shown that modifications to a SCADA system can be discovered

  19. CORSIKA modifications for faster background generation

    Science.gov (United States)

    Jero, Kyle

    2016-04-01

    CORSIKA is a simulation program for extensive air showers initiated by high energy cosmic particles. These air showers create the majority of the muons and neutrinos which neutrino that telescopes detect and are considered a background signature in searches for astrophysical neutrinos. This contribution will discuss changes to CORSIKA which allow for faster high energy background simulation. The theory, implementation, application, and performance of these modifications will be presented.

  20. CORSIKA modifications for faster background generation

    Directory of Open Access Journals (Sweden)

    Jero Kyle

    2016-01-01

    Full Text Available CORSIKA is a simulation program for extensive air showers initiated by high energy cosmic particles. These air showers create the majority of the muons and neutrinos which neutrino that telescopes detect and are considered a background signature in searches for astrophysical neutrinos. This contribution will discuss changes to CORSIKA which allow for faster high energy background simulation. The theory, implementation, application, and performance of these modifications will be presented.

  1. Food modification versus oral liquid nutrition supplementation.

    Science.gov (United States)

    Silver, Heidi J

    2009-01-01

    Oral liquid nutrition supplements (ONS) are widely used in community, residential and healthcare settings. ONS are intended for individuals whose nutrient requirements cannot be achieved by conventional diet or food modification, or for the management of distinctive nutrient needs resulting from specific diseases and/or conditions. ONS appear to be most effective in patients with a body mass index of

  2. Reproductive cloning combined with genetic modification.

    Science.gov (United States)

    Strong, C

    2005-11-01

    Although there is widespread opposition to reproductive cloning, some have argued that its use by infertile couples to have genetically related children would be ethically justifiable. Others have suggested that lesbian or gay couples might wish to use cloning to have genetically related children. Most of the main objections to human reproductive cloning are based on the child's lack of unique nuclear DNA. In the future, it may be possible safely to create children using cloning combined with genetic modifications, so that they have unique nuclear DNA. The genetic modifications could be aimed at giving such children genetic characteristics of both members of the couple concerned. Thus, cloning combined with genetic modification could be appealing to infertile, lesbian, or gay couples who seek genetically related children who have genetic characteristics of both members. In such scenarios, the various objections to human reproductive cloning that are based on the lack of genetic uniqueness would no longer be applicable. The author argues that it would be ethically justifiable for such couples to create children in this manner, assuming these techniques could be used safely.

  3. Integrated Microfluidics for Protein Modification Discovery*

    Science.gov (United States)

    Noach-Hirsh, Meirav; Nevenzal, Hadas; Glick, Yair; Chorni, Evelin; Avrahami, Dorit; Barbiro-Michaely, Efrat; Gerber, Doron; Tzur, Amit

    2015-01-01

    Protein post-translational modifications mediate dynamic cellular processes with broad implications in human disease pathogenesis. There is a large demand for high-throughput technologies supporting post-translational modifications research, and both mass spectrometry and protein arrays have been successfully utilized for this purpose. Protein arrays override the major limitation of target protein abundance inherently associated with MS analysis. This technology, however, is typically restricted to pre-purified proteins spotted in a fixed composition on chips with limited life-time and functionality. In addition, the chips are expensive and designed for a single use, making complex experiments cost-prohibitive. Combining microfluidics with in situ protein expression from a cDNA microarray addressed these limitations. Based on this approach, we introduce a modular integrated microfluidic platform for multiple post-translational modifications analysis of freshly synthesized protein arrays (IMPA). The system's potency, specificity and flexibility are demonstrated for tyrosine phosphorylation and ubiquitination in quasicellular environments. Unlimited by design and protein composition, and relying on minute amounts of biological material and cost-effective technology, this unique approach is applicable for a broad range of basic, biomedical and biomarker research. PMID:26276765

  4. Integrated Microfluidics for Protein Modification Discovery.

    Science.gov (United States)

    Noach-Hirsh, Meirav; Nevenzal, Hadas; Glick, Yair; Chorni, Evelin; Avrahami, Dorit; Barbiro-Michaely, Efrat; Gerber, Doron; Tzur, Amit

    2015-10-01

    Protein post-translational modifications mediate dynamic cellular processes with broad implications in human disease pathogenesis. There is a large demand for high-throughput technologies supporting post-translational modifications research, and both mass spectrometry and protein arrays have been successfully utilized for this purpose. Protein arrays override the major limitation of target protein abundance inherently associated with MS analysis. This technology, however, is typically restricted to pre-purified proteins spotted in a fixed composition on chips with limited life-time and functionality. In addition, the chips are expensive and designed for a single use, making complex experiments cost-prohibitive. Combining microfluidics with in situ protein expression from a cDNA microarray addressed these limitations. Based on this approach, we introduce a modular integrated microfluidic platform for multiple post-translational modifications analysis of freshly synthesized protein arrays (IMPA). The system's potency, specificity and flexibility are demonstrated for tyrosine phosphorylation and ubiquitination in quasicellular environments. Unlimited by design and protein composition, and relying on minute amounts of biological material and cost-effective technology, this unique approach is applicable for a broad range of basic, biomedical and biomarker research.

  5. Modification and Applications of Hydrophilic Polypropylene Membrane

    Science.gov (United States)

    Ariono, Danu; Kusuma Wardani, Anita

    2017-07-01

    Polypropylene (PP) is one of the most important polymers for microporous membrane due to its high void volume, well-controlled porosity, high thermal and chemical stability, and low cost. However, the hydrophobicity of PP becomes a limitation to broaden its applications. Furthermore, membrane fouling occurs more seriously on hydrophobic membranes than hydrophilic ones. To solve this problem, surface modifications have been developed to enhance PP membrane hydrophilicity without changing its bulk properties. Graft polymerization and plasma treatment are the most popular techniques for surface hydrophilization. Some studies showed that highly hydrophilic PP membranes with water contact angle less than 20° could be obtained by plasma treatment and graft polymerization. Furthermore, during plasma treatment, polar groups were formed on the PP membrane surface thus increased water uptake. To bring brief explanation on various research trends for PP modification, this paper provides a review of surface hydrophilization of microporous PP membrane, including plasma treatment and graft polymerization. The effects of surface modification on PP membrane performance such as porosity, water contact angle, and water flux are also discussed. In addition, the applications of modified PP membrane are presented as well.

  6. Improving active eigenvector assignment through passive modifications

    Science.gov (United States)

    Belotti, R.; Richiedei, D.

    2016-09-01

    Specifications on the dynamic behavior of feedback-controlled vibrating systems are often expressed in terms of its eigenstructure, i.e. eigenvalues and eigenvectors. The notion of controllability establishes the possibility to assign eigenvalues through state feedback, but it is not adequate to assure the assignment of arbitrary eigenvectors. Indeed, assignable eigenvectors are just those belonging to the allowable vector subspace, which depends on the physical properties of the vibrating system (mass, damping and stiffness matrices) and of the actuators. To overcome this limitation, this paper proposes a hybrid approach that exploits passive modification of the system physical parameters to modify the allowable subspace in such a way that it spans (or closely approximates) the desired eigenvectors. Then, once that the system modifications have been computed, standard techniques for control synthesis can be employed to compute the gains assigning the desired poles and the eigenvectors. The modification of the allowable subspace is cast in this work as a rank minimization problem, which can be efficiently tackled through semi-definite programming. The proposed method is numerically validated on a lumped parameter system, by proving that the assignment of eigenvectors by hybrid control is significantly enhanced compared with sole active control.

  7. Surface modification: advantages, techniques, and applications

    Energy Technology Data Exchange (ETDEWEB)

    Natesan, K.

    2000-03-01

    Adequate performance of materials at elevated temperatures is a potential problem in many systems within the chemical, petroleum, process, and power-generating industries. Degradation of materials occurs because of interaction between the structural material and the exposure environment. These interactions are generally undesired chemical reactions that can lead to accelerated wastage and alter the functional requirements and/or structural integrity of the materials. Therefore, material selection for high-temperature applications must be based not only on a material strength properties but also on resistance to the complex environments prevalent in the anticipated exposure environment. As plants become larger, the satisfactory performance and reliability of components play a greater role in plant availability and economics. However, system designers are becoming increasingly concerned with finding the least expensive material that will satisfactorily perform the design function for the desired service life. This present paper addresses the benefits of surface modification and identified several criteria for selection and application of modified surfaces in the power sector. A brief review is presented on potential methods for modification of surfaces, with the emphasis on coatings. In the final section of the paper, several examples address the requirements of different energy systems and surface modification avenues that have been applied to resolve the issues.

  8. Subtractive Structural Modification of Morpho Butterfly Wings.

    Science.gov (United States)

    Shen, Qingchen; He, Jiaqing; Ni, Mengtian; Song, Chengyi; Zhou, Lingye; Hu, Hang; Zhang, Ruoxi; Luo, Zhen; Wang, Ge; Tao, Peng; Deng, Tao; Shang, Wen

    2015-11-11

    Different from studies of butterfly wings through additive modification, this work for the first time studies the property change of butterfly wings through subtractive modification using oxygen plasma etching. The controlled modification of butterfly wings through such subtractive process results in gradual change of the optical properties, and helps the further understanding of structural optimization through natural evolution. The brilliant color of Morpho butterfly wings is originated from the hierarchical nanostructure on the wing scales. Such nanoarchitecture has attracted a lot of research effort, including the study of its optical properties, its potential use in sensing and infrared imaging, and also the use of such structure as template for the fabrication of high-performance photocatalytic materials. The controlled subtractive processes provide a new path to modify such nanoarchitecture and its optical property. Distinct from previous studies on the optical property of the Morpho wing structure, this study provides additional experimental evidence for the origination of the optical property of the natural butterfly wing scales. The study also offers a facile approach to generate new 3D nanostructures using butterfly wings as the templates and may lead to simpler structure models for large-scale man-made structures than those offered by original butterfly wings.

  9. Inorganic Surface Modification of Nonwoven Polymeric Substrates

    Science.gov (United States)

    Halbur, Jonathan Chandler

    In this study, atomic layer deposition (ALD), a vapor phase inorganic thin film deposition technique, is used to modify the surface of a range of industrially relevant polymers to enhance surface properties or impart additional functionalities. Several unique demonstrations of polymer surface modification are presented including uniform nanomaterial photodeposition to the surface of nonowoven fabrics and the first application of photocatalytic thin film coated nonwovens for advanced filtration of heavy metals from solution. Recent advances in polymer synthesis and processing technologies have resulted in the production of novel polymer systems with unique chemistries and sub-micron scale dimensions. As a result, advanced fiber systems have received much attention for potential use in a wide range of industrially and medically important applications such as advanced and selective filtration, catalysis, flexible electronics, and tissue engineering. However, tailoring the surface properties of the polymer is still needed in order to realize the full range of advanced applications, which can be difficult given the high complexity and non-uniformity of nonwoven polymeric structures. Uniform and controllable inorganic surface modification of nonwovens allows the introduction or modification of many crucial polymer properties with a wide range of application methods.

  10. Particle Distribution Modification by Low Amplitude Modes

    Energy Technology Data Exchange (ETDEWEB)

    White, R. B.; Gorelenkov, N.; Heidbrink, W. W.; Van Zeeland, M. A.

    2009-08-28

    Modification of a high energy particle distribution by a spectrum of low amplitude modes is investigated using a guiding center code. Only through resonance are modes effective in modifying the distribution. Diagnostics are used to illustrate the mode-particle interaction and to find which effects are relevant in producing significant resonance, including kinetic Poincare plots and plots showing those orbits with time averaged mode-particle energy transfer. Effects of pitch angle scattering and drag are studied, as well as plasma rotation and time dependence of the equilibrium and mode frequencies. A specific example of changes observed in a DIII-D deuterium beam distribution in the presence of low amplitude experimentally validated Toroidal Alfven (TAE) eigenmodes and Reversed Shear Alfven (RSAE) eigenmodes is examined in detail. Comparison with experimental data shows that multiple low amplitude modes can account for significant modification of high energy beam particle distributions. It is found that there is a stochastic threshold for beam profile modification, and that the experimental amplitudes are only slightly above this threshold.

  11. Post-production modification of industrial enzymes.

    Science.gov (United States)

    Minten, Inge J; Abello, Nicolas; Schooneveld-Bergmans, Margot E F; van den Berg, Marco A

    2014-01-01

    Industry has an increasing interest in the use of enzymes as environmentally friendly, highly efficient, and specific bio-catalysts. Enzymes have primarily evolved to function in aqueous environments at ambient temperature and pressure. These conditions however do not always correspond with industrial processes or applications, and only a small portion of all known enzymes are therefore suitable for industrial use. Protein engineering can sometimes be applied to convey more desirable properties to enzymes, such as increased stability, but is limited to the 20 naturally occurring amino acids or homologs thereof. Using post-production modification, which has the potential to combine desirable properties from the enzyme and the conjugated compounds, enzymes can be modified with both natural and synthetic molecules. This offers access to a myriad of possibilities for tuning the properties of enzymes. At this moment, however, the effects of post-production modification cannot yet be reliably predicted. The increasing number of applications will improve this so that the potential of this technology can be fully exploited. This review will focus on post-production modification of enzymes and its use and opportunities in industry.

  12. Modification Bituminous Binders Petroleum Resin (Based on C9 Fraction

    Directory of Open Access Journals (Sweden)

    Elena A. Chigorina

    2015-12-01

    Full Text Available The goal of the present study is to measure the basic parameters of a bituminous binder obtained by modification of the BND 60/90 binder with petroleum resin, for both dynamic and static modification modes.

  13. Reactive modification of polyesters and their blends

    Science.gov (United States)

    Wan, Chen

    2004-12-01

    As part of a broader research effort to investigate the chemical modification of polyesters by reactive processing a low molecular weight (MW) unsaturated polyester (UP) and a higher MW saturated polyester, polyethylene terephthalate (PET), alone or blended with polypropylene (PP) were melt processed in a batch mixer and continuous twin screw extruders. Modification was monitored by on-line rheology and the products were characterized primarily by off-line rheology, morphology and thermal analysis. Efforts were made to establish processing/property relationships and provide an insight of the accompanying structural changes. The overall response of the reactively modified systems was found to be strongly dependent on the component characteristics, blend composition, type and concentrations of reactive additives and processing conditions. The work concluded that UP can be effectively modified through reactive melt processing. Its melt viscosity and MW can be increased through chemical reactions between organic peroxides (POX) and chain unsaturation or between MgO and carboxyl/hydroxyl end groups. Reactive blending of PP/UP blends through peroxide modification gave finer and more uniform morphology than unreacted blends and at a given PP/UP weight ratio more thermoplastic elastomers-like rheological behavior. This is due to the continuously decreasing viscosity ratio of PP/UP towards unity by the competing reactions between POX and the blend components and formation of PP-UP copolymers which serve as in-situ compatibilizers to promote better interfacial adhesion. Kinetics of the competing reactions were analyzed through a developed model. In addition to POX concentration and mixing efficiency, rheology and morphology of UP/PP bends were significantly affected by the addition of inorganic and organic coagents. Addition of coagents such as a difunctional maleimide, MgO and/or an anhydride functionalized PP during reactive blending offers effective means for tailoring

  14. Chemical and structural effects of base modifications in messenger RNA

    Science.gov (United States)

    Harcourt, Emily M.; Kietrys, Anna M.; Kool, Eric T.

    2017-01-01

    A growing number of nucleobase modifications in messenger RNA have been revealed through advances in detection and RNA sequencing. Although some of the biochemical pathways that involve modified bases have been identified, research into the world of RNA modification -- the epitranscriptome -- is still in an early phase. A variety of chemical tools are being used to characterize base modifications, and the structural effects of known base modifications on RNA pairing, thermodynamics and folding are being determined in relation to their putative biological roles.

  15. 76 FR 6555 - Loan Guaranty Revised Loan Modification Procedures

    Science.gov (United States)

    2011-02-07

    ... receive any cash back from the modification. (b) If a loan fails to meet one or more of the conditions... AFFAIRS 38 CFR Part 36 RIN 2900-AN78 Loan Guaranty Revised Loan Modification Procedures AGENCY: Department... Veterans Affairs (VA) Loan Guaranty regulation related to modification of guaranteed housing loans...

  16. 49 CFR 237.133 - Supervision of repairs and modifications.

    Science.gov (United States)

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Supervision of repairs and modifications. 237.133... Supervision of repairs and modifications. Each repair or modification pursuant to this part shall be performed under the immediate supervision of a railroad bridge supervisor as defined in § 237.55 of this part...

  17. 48 CFR 53.243 - Contract modifications (SF 30).

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 2 2010-10-01 2010-10-01 false Contract modifications (SF... (CONTINUED) CLAUSES AND FORMS FORMS Prescription of Forms 53.243 Contract modifications (SF 30). SF 30 (Rev. 10/83), Amendment of Solicitation/Modification of Contract. SF 30 is prescribed for use in...

  18. 14 CFR 21.9 - Replacement and modification articles.

    Science.gov (United States)

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Replacement and modification articles. 21.9... CERTIFICATION PROCEDURES FOR PRODUCTS AND PARTS General § 21.9 Replacement and modification articles. (a) If a person knows, or should know, that a replacement or modification article is reasonably likely to...

  19. Sumoylation of Human Translationally Controlled Tumor Protein Is Important for Its Nuclear Transport

    OpenAIRE

    Gnanasekar Munirathinam; Kalyanasundaram Ramaswamy

    2012-01-01

    Translationally controlled tumor protein (TCTP) lacks nuclear bipartite localization signal sequence; yet TCTP is present abundantly in the nucleus. At present it is not known how TCTP gets transported to the nucleus. Sequence analyses showed that all TCTPs described to date have putative small ubiquitin-like modifier (SUMO) motifs. Since SUMO modification plays an important role in the nuclear transport of proteins, we evaluated whether SUMO motifs are important for transport of TCTP into th...

  20. Modification of the Stockholm Vascular Scale

    Energy Technology Data Exchange (ETDEWEB)

    Poole, K.; Elms, J.; Mason, H. [Health & Safety Laboratory, Buxton (United Kingdom)

    2006-09-15

    Staging hand-arm vibration syndrome (HAVS) depends upon accurate reporting of the extent and frequency of blanching attacks. Reporting may not be repeatable and not all individuals classifiable using the Stockholm Workshop Scale (SWS). For Department of Trade and Industry (Dti) coal miners' assessments, the SWS was modified to include a blanching score. Further modifications, which involve splitting Stage 2V into 'early' and 'late' have been proposed but the impact of this on classification has not been investigated. This ppaper investigates the impact of modifications in the SWS on HAVS classification using two modified scales. Two different cut-offs for defining 'frequent' blanching attacks ({>=} 3 or {>=} 7 attacks/week, respectively) were used. One hundred and sixty-five individuals were staged. Using the SWS, 58 and 31% of the population were unclassifiable using the two cut-offs, respectively. The modification splitting Stage 2V reduced the proportions that were unclassifiable to 2 and 9%, respectively, and increased those classified as Stage 2V. The cut-off for frequent attacks used (3 or 7) affected the proportion of individuals falling into the subdivisions of Stage 2 with 17 and 42% being classified as 2Vearly and 45 and 20% as 2Vlate, respectively. It was concluded that subdividing Stage 2V enables more individuals to be classified, but the proportion falling into each category is susceptible to the cut-off used for defining frequent attacks. Caution may need to be applied if this categorization is used to make decisions regarding fitness to work.

  1. Oxidative and other posttranslational modifications in extracellular vesicle biology.

    Science.gov (United States)

    Szabó-Taylor, Katalin; Ryan, Brent; Osteikoetxea, Xabier; Szabó, Tamás G; Sódar, Barbara; Holub, Marcsilla; Németh, Andrea; Pálóczi, Krisztina; Pállinger, Éva; Winyard, Paul; Buzás, Edit I

    2015-04-01

    Extracellular vesicles including exosomes, microvesicles and apoptotic vesicles, are phospholipid bilayer surrounded structures secreted by cells universally, in an evolutionarily conserved fashion. Posttranslational modifications such as oxidation, citrullination, phosphorylation and glycosylation play diverse roles in extracellular vesicle biology. Posttranslational modifications orchestrate the biogenesis of extracellular vesicles. The signals extracellular vesicles transmit between cells also often function via modulating posttranslational modifications of target molecules, given that extracellular vesicles are carriers of several active enzymes catalysing posttranslational modifications. Posttranslational modifications of extracellular vesicles can also contribute to disease pathology by e.g. amplifying inflammation, generating neoepitopes or carrying neoepitopes themselves.

  2. Dynamic Binary Modification Tools, Techniques and Applications

    CERN Document Server

    Hazelwood, Kim

    2011-01-01

    Dynamic binary modification tools form a software layer between a running application and the underlying operating system, providing the powerful opportunity to inspect and potentially modify every user-level guest application instruction that executes. Toolkits built upon this technology have enabled computer architects to build powerful simulators and emulators for design-space exploration, compiler writers to analyze and debug the code generated by their compilers, software developers to fully explore the features, bottlenecks, and performance of their software, and even end-users to extend

  3. Surface modification and functionalization of nanostructured carbons

    Directory of Open Access Journals (Sweden)

    A. Stanishevsky

    2009-12-01

    Full Text Available Purpose: Nanostructured carbon nanomaterials (e.g., nanocrystalline diamond films and particles, carbon nanotubes, carbon onions, fullerenes, etc. are being extensively explored for numerous biomedical applications in surgical implants, therapy, drug delivery, and biosensoring due to their interesting physical, chemical, and biological properties. Such applications of carbon nanomaterials often require specific surface functionality to be introduced for better integration of these materials with physiological environment. In the last decade, substantial progress has been made in the development of controllable surface modification methods and in the introduction of different functional groups on the surface of carbon nanomaterials.Design/methodology/approach: This paper briefly overviews the surface modification and functionalization approaches for various carbon nanomaterials, and it focuses on the plasma modification and functionalization of nanocrystalline diamond films, diamond nanoparticles, and carbon nanospheres. The results on the surface characterization using FTIR and XPS techniques, and the preliminary studies of cellular response to these modified carbon nanomaterials are presented and discussed.Findings: The results of surface modification of NCD films, detonation nanodiamonds, and carbon nanospheres, demonstrate the flexibility of nanocarbons to attain various surface functionality that can be adjusted for specific applications. It has been shown that neither of tested nanocarbon materials was cytotoxic in this study, although the attachement and proliferation of various cells was strongly affected by the specific type of surface functionalization.Research limitations/implications: At the present, it is not clear to what degree the available surface sites on NCD films or carbon nanoparticles can be occupied with functional groups. Furthermore, while there is clear selectivity of cellular response to H, O, and F surface

  4. Atmospheric pressure plasma for surface modification

    CERN Document Server

    Wolf, Rory A

    2012-01-01

    This Book's focus and intent is to impart an understanding of the practical application of atmospheric plasma for the advancement of a wide range of current and emerging technologies. The primary key feature of this book is the introduction of over thirteen years of practical experimental evidence of successful surface modifications by atmospheric plasma methods. It offers a handbook-based approach for leveraging and optimizing atmospheric plasma technologies which are currently in commercial use. It also offers a complete treatment of both basic plasma physics and industrial plasma process

  5. Modification of Ordinary Differential Equations MATLAB Solver

    Directory of Open Access Journals (Sweden)

    E. Cocherova

    2003-12-01

    Full Text Available Various linear or nonlinear electronic circuits can be described bythe set of ordinary differential equations (ODEs. The ordinarydifferential equations can be solved in the MATLAB environment inanalytical (symbolic toolbox or numerical way. The set of nonlinearODEs with high complexity can be usually solved only by use ofnumerical integrator (solver. The modification of ode23 MATLABnumerical solver has been suggested in this article for the applicationin solution of some special cases of ODEs. The main feature of thismodification is that the solution is found at every prescribed point,in which the special behavior of system is anticipated. Theextrapolation of solution is not allowed in those points.

  6. Magnetic zenith effect in ionospheric modifications

    Energy Technology Data Exchange (ETDEWEB)

    Gurevich, A.V.; Zybin, K.P.; Carlson, H.C.; Pedersen, T

    2002-12-09

    The theory of ionospheric modification for the beam of powerful radio emission directed along magnetic field lines is developed. Nonlinear process of beam self-focusing on striations is shown to determine strong amplification of heating and acceleration of plasma electrons. It results in a dramatic enhancement of optic emission from the magnetic zenith region in ionospheric F-layer. An excellent agreement between the theory and recent fundamental observations at HAARP facility (Alaska) [T. Pedersen et al., Geophys. Res. Lett. (2002), in press] is demonstrated.

  7. [TALE nuclease engineering and targeted genome modification].

    Science.gov (United States)

    Shen, Yan; Xiao, An; Huang, Peng; Wang, Wei-Ye; Zhu, Zuo-Yan; Zhang, Bo

    2013-04-01

    Artificial designer nucleases targeting specific DNA sequences open up a new field for reverse genetics study. The rapid development of engineered endonucleases (EENs) enables targeted genome modification theoretically in any species. The construction of transcription activator-like effector nucleases (TALENs) is simpler with higher specificity and less toxicity than zinc-finger nucleases (ZFNs). Here, we summarized the recent progresses and prospects of TALEN technology, with an emphasis on its structure, function, and construction strategies, as well as a collection of species and genes that have been successfully modified by TALENs, especially the application in zebrafish.

  8. Modified Kriging: evaluation, modification, and recommendations

    Energy Technology Data Exchange (ETDEWEB)

    Schultz, C.A.; Myers, S.C.

    1997-09-01

    This report describes modifications to a technique developed by Sandia National Laboratory (SNL) to predict general corrections (traveltime, etc.) for a geographic grid only sparsely covered by calibration points (Hipp and Young, 1997). SNL has worked to create a modified version of linear prediction (kriging) based on the idea of blending the surface back to zero at some distance from the points or, in other words, by damping the solution through the damping of the input data points. LLNL has been working with SNL to evaluate Modified Kriging. This report documents our evaluation of the technique and our resulting recommendations to SNL.

  9. Bt toxin modification for enhanced efficacy.

    Science.gov (United States)

    Deist, Benjamin R; Rausch, Michael A; Fernandez-Luna, Maria Teresa; Adang, Michael J; Bonning, Bryony C

    2014-10-22

    Insect-specific toxins derived from Bacillus thuringiensis (Bt) provide a valuable resource for pest suppression. Here we review the different strategies that have been employed to enhance toxicity against specific target species including those that have evolved resistance to Bt, or to modify the host range of Bt crystal (Cry) and cytolytic (Cyt) toxins. These strategies include toxin truncation, modification of protease cleavage sites, domain swapping, site-directed mutagenesis, peptide addition, and phage display screens for mutated toxins with enhanced activity. Toxin optimization provides a useful approach to extend the utility of these proteins for suppression of pests that exhibit low susceptibility to native Bt toxins, and to overcome field resistance.

  10. On cavity modification of stimulated Raman scattering

    CERN Document Server

    Matsko, A B; Letargat, R J; Ilchenko, V S; Maleki, L

    2003-01-01

    We study theoretically stimulated Raman scattering (SRS) in a nonlinear dielectric microcavity and compare SRS thresholds for the cavity and the bulk material it is made of. We show that cavity SRS enhancement results solely from the intensity build up in the cavity and from the differences of the SRS dynamics in free and confined space. There is no significant modification of the Raman gain due to cavity QED effects. We show that the SRS threshold depends significantly on the nature of the dominating cavity decay as well as on the coupling technique with the cavity used for SRS measurements.

  11. Characteristic of involute slope modification of asymmetric spur gear

    Institute of Scientific and Technical Information of China (English)

    DENG Xiao-he; HUA Lin; HAN Xing-hui

    2015-01-01

    The meshing characteristic of asymmetric involute spur gear was studied, the equations of the geometric shape of the asymmetric gear for both sides were deduced, and the equations of contact ratio and the key points of contact were also obtained. Meanwhile, an involute slope modification method considering the effects of static transmission errors was proposed based on the meshing properties. The characteristic of the involute slope modification was analyzed by changing different modification parameters. The mesh stiffness and synthetic mesh stiffness of unmodified and modified asymmetric spur gears were investigated. Furthermore, the spectrums of synthetic mesh stiffness under different modification parameters were compared. Research results showed that the modification parameters influence the meshing performance of gear pairs, and the proposed modification method was feasible to improve the transmission performance of gear pairs with appropriate modification parameters.

  12. Chemical modification of soybean oil for lubricant

    Energy Technology Data Exchange (ETDEWEB)

    Duan Shijie; Mao Zongqiang [Tsinghua Univ., INET, BJ (China)

    2008-07-01

    This paper presents a series of structural modifications of soybean oils for lubricant. The reaction was monitored and products were confirmed by NMR and FTIR. The structural modification is carried out in four stages, (1) synthesis of soybean oil isooctyl ester from soybean oil; (2) synthesis of epoxy-soybean oil isooctyl ester from soybean oil isooctyl ester; (3) synthesis of hydroxylated products from epoxy soybean oil isooctyl ester with fatty acid; (4) esterification of the hydroxylated product with anhydride. Rheological behaviour of the products was measured. Pour points of the products (3) were observed as low as -24 C (lauric acid) and -15 C (isooctanoic acid) respectively. When the hydroxyl groups in the products were esterified with and acid anhydride, the pour points were became higher, which were -21 C (lauric acid) and -6 C (isooctanoic acid) without pour point depressant, and -27 C (lauric acid) and -24 C (isooctanoic acid) with 1% of pour point depressant respectively. The products have suitable viscosity and viscosity index, and the viscosity indices are all above 100. (orig.)

  13. MODIFICATION OF DELTA FOR CHOOSER OPTIONS

    Directory of Open Access Journals (Sweden)

    Marek Ďurica

    2015-09-01

    Full Text Available Correctly used financial derivatives can help investors increase their expected returns and minimize their exposure to risk. To ensure the specific needs of investors, a large number of different types of non-standard exotic options is used. Chooser option is one of them. It is an option that gives its holder the right to choose at some predetermined future time whether the option will be a standard call or put with predetermined strike price and maturity time. Although the chooser options are more expensive than standard European-style options, in many cases they are a more suitable instrument for investors in hedging their portfolio value. For an effective use of the chooser option as a hedging instrument, it is necessary to check the values of the Greek parameters delta and gamma for the options. Especially, if the value of the parameter gamma is too large, hedging of the portfolio value using only parameter delta is insufficient and brings high transaction costs because the portfolio has to be reviewed relatively often. Therefore, in this article, a modification of delta-hedging as well as using the value of parameter gamma is suggested. Error of the delta modification is analyzed and compared with the error of widely used parameter delta. Typical patterns for the modified hedging parameter variation with various time to choose time for chooser options are also presented in this article.

  14. Detection of thiol modifications by hydrogen sulfide.

    Science.gov (United States)

    Williams, E; Pead, S; Whiteman, M; Wood, M E; Wilson, I D; Ladomery, M R; Teklic, T; Lisjak, M; Hancock, J T

    2015-01-01

    Hydrogen sulfide (H2S) is an important gasotransmitter in both animals and plants. Many physiological events, including responses to stress, have been suggested to involve H2S, at least in part. On the other hand, numerous responses have been reported following treatment with H2S, including changes in the levels of antioxidants and the activities of transcription factors. Therefore, it is important to understand and unravel the events that are taking place downstream of H2S in signaling pathways. H2S is known to interact with other reactive signaling molecules such as reactive oxygen species (ROS) and nitric oxide (NO). One of the mechanisms by which ROS and NO have effects in a cell is the modification of thiol groups on proteins, by oxidation or S-nitrosylation, respectively. Recently, it has been reported that H2S can also modify thiols. Here we report a method for the determination of thiol modifications on proteins following the treatment with biological samples with H2S donors. Here, the nematode Caenorhabditis elegans is used as a model system but this method can be used for samples from other animals or plants.

  15. Surface modification of barite nanoparticles using stearate

    Institute of Scientific and Technical Information of China (English)

    LI Lin-lin; HANG Jian-zhong; SHI Li-yi

    2009-01-01

    In this study,the barite nanoparticles were successfully modified with stearate and the influence of stearate addition on the performance of barite nanoparticles was systematically investigated.The products were characterized by activating factor analysis,contact angle test,surface energy calculation,sedimentation rate calculation,rheological measurement,and FT-IR analysis,etc.As the quantity of added stearate increased,both the activating factor and contact angle of barite nanoparticles increased first then decreased.When the stearate content was 5% of the mass of barite nanoparticles,the activating factor and water contact angle of modified particles reached maximum value,97% and 126~ respectively.At this time,the sedimentation rate reached minimum,and so did the surface energy.The rheological test reveals that the viscosity of modified barite nanoparticles/ petronol system decreases greatly,indicating the surface performance of barite nanoparticles has changed from hydrophilicity to lipophilicity after modification.C=O and COO stretching vibration peaks were found in the FT-IR spectra,which proves that the stearate has combined onto the surface of barite nanoparticles.Finally,according to the zeta potential result of unmodified barite,the possible modification mechanism was provided.

  16. STIFFNESS MODIFICATION OF COTTON IN CHITOSAN TREATMENT

    Directory of Open Access Journals (Sweden)

    CAMPOS Juan

    2017-05-01

    Full Text Available Chitosan is a biopolymer obtained from chitin, and among their most important aspects highlights its applications in a lot of industrial sectors due to its intrinsic properties, especially in the textile sector. In the last years, chitosan is widely used in the cotton and wool finishing processes due to its bond between them and its properties as an antifungical and antimicrobial properties. In this paper three different molecular weight chitosan are used in the finishing process of cotton to evaluate its influence in the surface properties modification. In order to evaluate the effect of the treatment with chitosan, flexural stiffness test is performed in warp and weft direction, and then the total value is calculated. The cotton fabric is treated with 5 g/L of different types of chitosan in an impregnation bath. This study shows the extent of surface properties modification of the cotton provided by three types of chitosan treatment. The results show that all types of chitosan modify the cotton flexural rigidity properties but the one which modifies it in a relevant manner is chitosan originated from shrimps. Chitosan, textile, flexural stiffnes, chitin, cotton.

  17. Wetting and adsorption modification in the system

    Directory of Open Access Journals (Sweden)

    Yuliya Bogdanova

    2015-09-01

    Full Text Available Regularities of wetting and adsorption modification of surfaces of continual membranes made from highly permeable glassy polymers poly[1-(trimethylsilyl-1-propyne] (PTMSP and poly(4-methyl-2-pentyn (PMP with aqueous ethanol solutions and alcohol solutions containing organic dyes (Solvent Blue 35 and Remazol Brilliant Blue were investigated. Isotherms of stress wetting of polymer membrane surface by etanol solutions were found out to have maximums in the range of concentrations corresponding to the beginning of liquid sorption into the membrane and polymer swelling. Thus, the principal possibility of optimization of nanofiltration experiments by liquid wetting angle measurements on continuous polymer membrane surfaces was shown. The presence of the dye was shown not to affect PMP wetting. But in the case of PTMSP, it leads to shear of the maximum of stress wetting isotherms to the range of higher concentrations. It was found out the effectiveness of the adsorption surface modification of continuous polymer membrane surfaces by ethanol solutions containing dyes does not dependent on chemical nature of the dye. At the same time, there are different trends in the energy characteristics of the membrane surface.

  18. Targeting aging for disease modification in osteoarthritis.

    Science.gov (United States)

    Collins, John A; Diekman, Brian O; Loeser, Richard F

    2017-09-27

    Age is a key risk factor for the development of osteoarthritis and age-related changes within the joint might represent targets for therapy. The recent literature was reviewed to find studies that provide new insight into the role of aging in osteoarthritis, with a focus on the potential for disease modification. Preclinical studies using isolated cells and animal models provide evidence that two hallmarks of aging (cellular senescence and mitochondrial dysfunction) contribute to the development of osteoarthritis. Senescent cells secrete pro-inflammatory mediators and matrix degrading enzymes, and killing these cells with 'senolytic' compounds has emerged as a potential disease-modifying therapy. Mitochondrial dysfunction is associated with increased levels of reactive oxygen species (ROS) that can promote osteoarthritis by disrupting homeostatic intracellular signaling. Reducing ROS production in the mitochondria, stimulating antioxidant gene expression through Nrf2 activation, or inhibiting specific redox-sensitive signaling proteins represent additional approaches to disease modification in osteoarthritis that require further investigation. Although no human clinical trials for osteoarthritis have specifically targeted aging, preclinical studies suggest that targeting cellular senescence and/or mitochondrial dysfunction and the effects of excessive ROS may lead to novel interventions that could slow the progression of osteoarthritis.

  19. Electron beam modification of vanadium dioxide oscillators

    Energy Technology Data Exchange (ETDEWEB)

    Belyaev, Maksim; Velichko, Andrey; Putrolaynen, Vadim; Perminov, Valentin; Pergament, Alexander [Petrozavodsk State University, Petrozavodsk (Russian Federation)

    2017-03-15

    The paper presents the results of a study of electron-beam modification (EBM) of VO{sub 2}-switch I-V curve threshold parameters and the self-oscillation frequency of a circuit containing such a switching device. EBM in vacuum is reversible and the parameters are restored when exposed to air at pressure of 150 Pa. At EBM with a dose of 3 C cm{sup -2}, the voltages of switching-on (V{sub th}) and off (V{sub h}), as well as the OFF-state resistance R{sub off}, decrease down to 50% of the initial values, and the oscillation frequency increases by 30% at a dose of 0.7 C cm{sup -2}. Features of physics of EBM of an oscillator are outlined considering the contribution of the metal and semiconductor phases of the switching channel. Controlled modification allows EBM forming of switches with preset parameters. Also, it might be used in artificial oscillatory neural networks for pattern recognition based on frequency shift keying. (copyright 2017 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  20. Posttranslational Modifications and the Immunogenicity of Biotherapeutics

    Directory of Open Access Journals (Sweden)

    Roy Jefferis

    2016-01-01

    Full Text Available Whilst the amino acid sequence of a protein is determined by its gene sequence, the final structure and function are determined by posttranslational modifications (PTMs, including quality control (QC in the endoplasmic reticulum (ER and during passage through the Golgi apparatus. These processes are species and cell specific and challenge the biopharmaceutical industry when developing a production platform for the generation of recombinant biologic therapeutics. Proteins and glycoproteins are also subject to chemical modifications (CMs both in vivo and in vitro. The individual is naturally tolerant to molecular forms of self-molecules but nonself variants can provoke an immune response with the generation of anti-drug antibodies (ADA; aggregated forms can exhibit enhanced immunogenicity and QC procedures are developed to avoid or remove them. Monoclonal antibody therapeutics (mAbs are a special case because their purpose is to bind the target, with the formation of immune complexes (ICs, a particular form of aggregate. Such ICs may be removed by phagocytic cells that have antigen presenting capacity. These considerations may frustrate the possibility of ameliorating the immunogenicity of mAbs by rigorous exclusion of aggregates from drug product. Alternate strategies for inducing immunosuppression or tolerance are discussed.

  1. Heavy flavored jet modification in CMS

    CERN Document Server

    Jung, Kurt

    2015-01-01

    The energy loss of jets in heavy-ion collisions is expected to depend on the flavor of the fragmenting parton. Thus, measurements of jet quenching as a function of flavor place powerful constraints on the thermodynamical and transport properties of the hot and dense medium. Measurements of the nuclear modification factors of the heavy-flavor-tagged jets (from charm and bottom quarks) in both PbPb and pPb collisions can quantify such energy loss effects. Specifically, pPb measurements provide crucial insights into the behavior of the cold nuclear matter effect, which is required to fully understand the hot and dense medium effects on jets in PbPb collisions. In this talk, we present the heavy flavor jet spectra and measurements of the nuclear modification factors in both PbPb and pPb as a function of transverse momentum and pseudorapidity, using the high statistics pp, pPb and PbPb data taken in 2011 and 2013. Finally, we also will present a proposal for c-jet tagging methodology to be used for the upcoming hi...

  2. The NEDD8 modification pathway in plants

    Directory of Open Access Journals (Sweden)

    Claus eSchwechheimer

    2014-03-01

    Full Text Available NEDD8, in plants and yeasts also known as RELATED TO UBIQUITIN (RUB, is an evolutionarily conserved 76 amino acid protein highly related to ubiquitin. Like ubiquitin, NEDD8 can be conjugated to and deconjugated from target proteins, but unlike ubiquitin, NEDD8 has not been reported to form chains similar to the different polymeric ubiquitin chains that have a role in a diverse set of cellular processes. NEDD8-modification is best known as a posttranslational modification of the cullin subunits of cullin-RING E3 ubiquitin ligases. In this context, structural analyses have revealed that neddylation induces a conformation change of the cullin that brings the ubiquitylation substrates into proximity of the interacting E2 conjugating enzyme. In turn, NEDD8 deconjugation destabilizes the cullin RING ligase complex allowing for the exchange of substrate recognition subunits via the exchange factor CAND1. In plants, components of the neddylation and deneddylation pathway were identified based on mutants with defects in auxin and light responses and the characterization of these mutants has been instrumental for the elucidation of the neddylation pathway. More recently, there has been evidence from animal and plant systems that NEDD8 conjugation may also regulate the behavior or fate of non-cullin substrates in a number of ways. Here, the current knowledge on NEDD8 processing, conjugation and deconjugation is presented, where applicable, in the context of specific signaling pathways from plants.

  3. Studing cranial vault modifications in ancient Mesoamerica.

    Science.gov (United States)

    Tiesler, Vera

    2012-01-01

    The artificial modification of infant cranial vaults through massages or by means of constriction and compression devices constitutes a readily visible, permanent body modification that has been employed cross-culturally to express identity, ethnicity, beauty, status and gender. For those ancient societies that staged head shaping, these cultural correlates may be ascertained by examining cranial shapes together with other data sets from the archaeological record. Studies of skulls modified for cultural reasons also provide important clues for understanding principles in neural growth and physiopathological variation in cranial expansion. This paper focuses on head shaping techniques in Mesoamerica, where the practice was deeply rooted and widespread before the European conquest. It provides a comprehensive review of the Mesoamericanistic research on shaping techniques, implements and taxonomies. An up-dated, interdisciplinary examination of the physiological implications and the cultural meanings of artificially produced head shapes in different times and culture areas within Mesoamerica leads to a discussion of the scope, caveats, and future directions involved in this kind of research in the region and beyond.

  4. Modification methods for poly(arylsulfone) membranes: A mini-review focusing on surface modification

    NARCIS (Netherlands)

    Nady, N.; Franssen, M.C.R.; Zuilhof, H.; Mohy Eldin, M.S.; Boom, R.M.; Schroën, C.G.P.H.

    2011-01-01

    Surface modification of membranes is thought to be equally important to the membrane industry as membrane material and process development; surface functionalization has already become a key technology, the major aims being performance improvement (flux and selectivity) by reduction of unwanted prot

  5. Modification-specific proteomics: strategies for characterization of post-translational modifications using enrichment techniques

    DEFF Research Database (Denmark)

    Zhao, Yingming; Jensen, Ole N

    2009-01-01

    substrates and their PTM sites is key to dissection of PTM-mediated cellular processes. The past several years have seen a tremendous progress in developing MS-based proteomics technologies for global PTM analysis, including numerous studies of yeast and other microbes. Modification-specific enrichment...

  6. 40 CFR 53.14 - Modification of a reference or equivalent method.

    Science.gov (United States)

    2010-07-01

    ... intended modification of the method, including but not limited to modifications of design or construction... intended modification. (2) A brief statement of the applicant's belief that the modification will, will...

  7. Mitochondrial Modification Techniques and Ethical Issues

    Directory of Open Access Journals (Sweden)

    Lucía Gómez-Tatay

    2017-02-01

    Full Text Available Current strategies for preventing the transmission of mitochondrial disease to offspring include techniques known as mitochondrial replacement and mitochondrial gene editing. This technology has already been applied in humans on several occasions, and the first baby with donor mitochondria has already been born. However, these techniques raise several ethical concerns, among which is the fact that they entail genetic modification of the germline, as well as presenting safety problems in relation to a possible mismatch between the nuclear and mitochondrial DNA, maternal mitochondrial DNA carryover, and the “reversion” phenomenon. In this essay, we discuss these questions, highlighting the advantages of some techniques over others from an ethical point of view, and we conclude that none of these are ready to be safely applied in humans.

  8. Mitochondrial Modification Techniques and Ethical Issues

    Science.gov (United States)

    Gómez-Tatay, Lucía; Hernández-Andreu, José M.; Aznar, Justo

    2017-01-01

    Current strategies for preventing the transmission of mitochondrial disease to offspring include techniques known as mitochondrial replacement and mitochondrial gene editing. This technology has already been applied in humans on several occasions, and the first baby with donor mitochondria has already been born. However, these techniques raise several ethical concerns, among which is the fact that they entail genetic modification of the germline, as well as presenting safety problems in relation to a possible mismatch between the nuclear and mitochondrial DNA, maternal mitochondrial DNA carryover, and the “reversion” phenomenon. In this essay, we discuss these questions, highlighting the advantages of some techniques over others from an ethical point of view, and we conclude that none of these are ready to be safely applied in humans. PMID:28245555

  9. Autoantibodies to Posttranslational Modifications in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Agata N. Burska

    2014-01-01

    Full Text Available Autoantibodies have been associated with human pathologies for a long time, particularly with autoimmune diseases (AIDs. Rheumatoid factor (RF is known since the late 1930s to be associated with rheumatoid arthritis (RA. The discovery of anticitrullinated protein antibodies in the last century has changed this and other posttranslational modifications (PTM relevant to RA have since been described. Such PTM introduce neoepitopes in proteins that can generate novel autoantibody specificities. The recent recognition of these novel specificities in RA provides a unique opportunity to understand human B-cell development in vivo. In this paper, we will review the three of the main classes of PTMs already associated with RA: citrullination, carbamylation, and oxidation. With the advancement of research methodologies it should be expected that other autoantibodies against PTM proteins could be discovered in patients with autoimmune diseases. Many of such autoantibodies may provide significant biomarker potential.

  10. Modification of graphene by ion irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Bukowska, Hanna; Akcoeltekin, Sevilay; El Kharrazi, Mourad; Schleberger, Marika [Universitaet Duisburg-Essen, Fakultaet fuer Physik, Duisburg (Germany); Osmani, Orkhan [Universitaet Duisburg-Essen, Fakultaet fuer Physik, Duisburg (Germany); Technische Universitaet Kaiserslautern, Fachbereich Physik, Gottlieb-Daimler-Strasse, Gebaeude 47, 67663 Kaiserslautern (Germany)

    2010-07-01

    Ion irradiation can be used to modify surfaces on the nanometer scale. We investigate graphene on different insulator (SrTiO{sub 3}, TiO{sub 2}, and Al{sub 2}O{sub 3}) and semiconductor (SiO{sub 2}) substrates. The bombardment of those target surfaces with swift heavy ions under grazing angle of incidence creates chains of nanodots on the substrate and folds graphene to typical origami-like structures. The shape of the folded graphene seems to depend on the length of the tracks. The length can be controlled by the angle of incidence. From the analysis of atomic force microscopy measurements, we classify the different types of modifications, with the aim to determine the relationship between chain length and origami shape. Further more we want to develop a theoretical understanding of the physical processes leading to the folding.

  11. Genome modification by CRISPR/Cas9.

    Science.gov (United States)

    Ma, Yuanwu; Zhang, Lianfeng; Huang, Xingxu

    2014-12-01

    Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein (Cas)9-mediated genome modification enables us to edit the genomes of a variety of organisms rapidly and efficiently. The advantages of the CRISPR-Cas9 system have made it an increasingly popular genetic engineering tool for biological and therapeutic applications. Moreover, CRISPR-Cas9 has been employed to recruit functional domains that repress/activate gene expression or label specific genomic loci in living cells or organisms, in order to explore developmental mechanisms, gene expression regulation, and animal behavior. One major concern about this system is its specificity; although CRISPR-Cas9-mediated off-target mutation has been broadly studied, more efforts are required to further improve the specificity of CRISPR-Cas9. We will also discuss the potential applications of CRISPR-Cas9.

  12. Modifications to the silver physical developer.

    Science.gov (United States)

    Burow, David; Seifert, Donald; Cantu, Antonio A

    2003-09-01

    The silver physical developer is currently the most successful reagent used for visualizing the water-insoluble components (e.g., lipids) of latent prints on porous surfaces. It is normally used after the amino acid visualizing reagents (e.g., ninhydrin and DFO) are used. This work found that the performance of the current formulation of silver physical developer is strongly reduced when the water used is changed from the usual distilled water to the more purified reverse osmosis/deionized (RO/DI) water. Based on numerous experiments involving the systematic variation of the component concentrations, the performance was restored and even improved by reducing the concentration of all the components (except that of the ferric salt) and by including malic acid in the formulation. These modifications resulted in a new silver physical developer formulation that performs as well as or better than the current formulation and is less expensive to make.

  13. Epigenetic Modifications of Major Depressive Disorder

    Directory of Open Access Journals (Sweden)

    Kathleen Saavedra

    2016-08-01

    Full Text Available Major depressive disorder (MDD is a chronic disease whose neurological basis and pathophysiology remain poorly understood. Initially, it was proposed that genetic variations were responsible for the development of this disease. Nevertheless, several studies within the last decade have provided evidence suggesting that environmental factors play an important role in MDD pathophysiology. Alterations in epigenetics mechanism, such as DNA methylation, histone modification and microRNA expression could favor MDD advance in response to stressful experiences and environmental factors. The aim of this review is to describe genetic alterations, and particularly altered epigenetic mechanisms, that could be determinants for MDD progress, and how these alterations may arise as useful screening, diagnosis and treatment monitoring biomarkers of depressive disorders.

  14. TALEN utilization in rice genome modifications.

    Science.gov (United States)

    Li, Ting; Liu, Bo; Chen, Chih Ying; Yang, Bing

    2014-08-15

    Transcription activator-like effector nucleases (TALENs), the newly developed and powerful genetic tools for precise genome editing, are fusion proteins of TAL effectors as DNA binding domains and the cleavage domain of FokI endonuclease. As a pair, the central repeat regions of TALENs determine the DNA binding specificity for the two sub-target sites; and the dimeric non-specific FokI cleavage domains cause a DNA double strand break (DSB) between the bound sequences. In vivo, cells repair the DSBs through either non-homologous end joining (NHEJ) pathway or homologous recombination (HR) pathway. Various methods have been developed for easy and fast assembly of TALEN genes for their utilization in a variety of eukaryotic cells or organisms. Here we present a TALEN-based rice genome modification protocol including constructing modularly assembled TALENs, rice transformation, and mutant screening.

  15. A ``fractal'' modification of Torricelli's formula

    Science.gov (United States)

    Maramathas, Athanasios J.; Boudouvis, Andreas G.

    2010-03-01

    A modification is proposed of Torricelli’s (1608-1647) formula for the velocity of water discharging from a small hole at the bottom of a large tank filled with fractal solid material. The new formula takes proper account of the mechanical energy losses due to flow in the solid matrix, thus expanding the area of validity of the classical Torricelli’s formula. Moreover, it offers a convenient alternative to Darcy’s law for estimating the discharge rate from an aquifer. The new formula was derived from laboratory experiments, with a low-Reynolds number discharge flow (Darcian flow). It was tested in a natural karst aquifer where the flow is non-Darcian, at Almiros spring on the island of Crete (Greece). In both cases, the predictive capability of the modified formula is established.

  16. Autoantibodies to Posttranslational Modifications in Rheumatoid Arthritis

    Science.gov (United States)

    Burska, Agata N.; Hunt, Laura; Strollo, Rocky; Ryan, Brent J.; Vital, Ed; Nissim, Ahuva; Winyard, Paul G.; Emery, Paul; Ponchel, Frederique

    2014-01-01

    Autoantibodies have been associated with human pathologies for a long time, particularly with autoimmune diseases (AIDs). Rheumatoid factor (RF) is known since the late 1930s to be associated with rheumatoid arthritis (RA). The discovery of anticitrullinated protein antibodies in the last century has changed this and other posttranslational modifications (PTM) relevant to RA have since been described. Such PTM introduce neoepitopes in proteins that can generate novel autoantibody specificities. The recent recognition of these novel specificities in RA provides a unique opportunity to understand human B-cell development in vivo. In this paper, we will review the three of the main classes of PTMs already associated with RA: citrullination, carbamylation, and oxidation. With the advancement of research methodologies it should be expected that other autoantibodies against PTM proteins could be discovered in patients with autoimmune diseases. Many of such autoantibodies may provide significant biomarker potential. PMID:24782594

  17. Machining, joining and modifications of advanced materials

    CERN Document Server

    Altenbach, Holm

    2016-01-01

    This book presents the latest advances in mechanical and materials engineering applied to the machining, joining and modification of modern engineering materials. The contributions cover the classical fields of casting, forming and injection moulding as representative manufacturing methods, whereas additive manufacturing methods (rapid prototyping and laser sintering) are treated as more innovative and recent technologies that are paving the way for the manufacturing of shapes and features that traditional methods are unable to deliver. The book also explores water jet cutting as an innovative cutting technology that avoids the heat build-up typical of classical mechanical cutting. It introduces readers to laser cutting as an alternative technology for the separation of materials, and to classical bonding and friction stir welding approaches in the context of joining technologies. In many cases, forming and machining technologies require additional post-treatment to achieve the required level of surface quali...

  18. Controller modification applied for active fault detection

    DEFF Research Database (Denmark)

    Niemann, Hans Henrik; Stoustrup, Jakob; Poulsen, Niels Kjølstad

    2014-01-01

    This paper is focusing on active fault detection (AFD) for parametric faults in closed-loop systems. This auxiliary input applied for the fault detection will also disturb the external output and consequently reduce the performance of the controller. Therefore, only small auxiliary inputs are used...... with the result that the detection and isolation time can be long. In this paper it will be shown, that this problem can be handled by using a modification of the feedback controller. By applying the YJBK-parameterization (after Youla, Jabr, Bongiorno and Kucera) for the controller, it is possible to modify...... the feedback controller with a minor effect on the external output in the fault free case. Further, in the faulty case, the signature of the auxiliary input can be optimized. This is obtained by using a band-pass filter for the YJBK parameter that is only effective in a small frequency range where...

  19. Gold Nanowires and Their Chemical Modifications

    Science.gov (United States)

    Häkkinen, Hannu; Barnett, Robert N.; Landman, Uzi

    2000-03-01

    Atomic structure, electronic structure, and ballistic transport in thin gold nanowires at their final stages before break-up, recently imaged by high-resolution electron microscopy,(H. Ohnishi et al., Nature 395), 780 (1998) are investigated with density functional simulations.(H. Häkkinen et al., J. Phys. Chem. B 103), 8814 (1999) We discuss stretching mechanisms leading to elongated chain-like structures showing dimerization akin to a Peierls transition and retaining conductance close to unity for stretching lengths that exceed considerably a typical interatomic bond length. We also demonstrate a chemical modification of the wire via adsorption of a methyl thiol molecule and discuss its effects on the structure and conductance.

  20. A study of microbial profile modification

    Energy Technology Data Exchange (ETDEWEB)

    Bae, J.H.; Lee, H.O.

    1995-12-31

    A microbial profile modification method using spores was investigated. A halotolerant, spore-forming, biopolymer-producing mesophile was used in Berea cores with a specifically formulated nutrient package to reduce the permeability of the rock. The degree of permeability reduction varied widely depending on the stimulation protocols used. The incubation period had a significant impact on permeability reduction, and there appeared to be an optimum incubation time for maximum permeability reduction. The reduction persisted for many PV of brine injection and appeared very stable. For our microbes used in this study, the permeability reduction was about the same when the NaCl concentration was above 2 wt% in the range from 0 wt% to 10 wt%.

  1. Genetic Modification of Baculovirus Expression Vectors

    Institute of Scientific and Technical Information of China (English)

    Shu-fen Li; Hua-lin Wang; Zhi-hong Hu; Fei Deng

    2012-01-01

    As a protein expression vector,the baculovirus demonstrates many advantages over other vectors.With the development of biotechnology,baculoviral vectors have been genetically modified to facilitate high level expression of heterologous proteins in both insect and mammalian cells.These modifications include utilization of different promoters and signal peptides,deletion or replacement of viral genes for increasing protein secretion,integration of polycistronic expression cassette for producing protein complexes,and baculovirus pseudotyping,promoter accommodation or surface display for enhancing mammalian cell targeting gene delivery.This review summarizes the development and the current state of art of the baculovirus expression system.Further development of baculovirus expression systems will make them even more feasible and accessible for advanced applications.

  2. Posttranslational Protein Modifications in Plant Metabolism.

    Science.gov (United States)

    Friso, Giulia; van Wijk, Klaas J

    2015-11-01

    Posttranslational modifications (PTMs) of proteins greatly expand proteome diversity, increase functionality, and allow for rapid responses, all at relatively low costs for the cell. PTMs play key roles in plants through their impact on signaling, gene expression, protein stability and interactions, and enzyme kinetics. Following a brief discussion of the experimental and bioinformatics challenges of PTM identification, localization, and quantification (occupancy), a concise overview is provided of the major PTMs and their (potential) functional consequences in plants, with emphasis on plant metabolism. Classic examples that illustrate the regulation of plant metabolic enzymes and pathways by PTMs and their cross talk are summarized. Recent large-scale proteomics studies mapped many PTMs to a wide range of metabolic functions. Unraveling of the PTM code, i.e. a predictive understanding of the (combinatorial) consequences of PTMs, is needed to convert this growing wealth of data into an understanding of plant metabolic regulation.

  3. Selective Surface Modification on Lubricant Retention

    Science.gov (United States)

    Jiang, Yu; Suvanto, Mika; Pakkanen, Tapani A.

    2016-11-01

    While surface patterns are effective in improving tribological properties, nevertheless they alter the surface wettability, which will in turn affect the surface-lubricant interactions. When there is a shortage of lubricant on a patterned surface, the lubricant stored inside the cavities will be extracted to compensate the surface lubricant dissipation. Additionally, the lubricant retention effect provided by the cavities is competing with the release of the lubricant. With weak surface-lubricant interaction, the retention is limited. Therefore, the lubrication will have a sudden failure, giving a dramatic transition to abrasive wear. To improve the performance of polar lubricants on hydrophobic polymer surfaces, both topographical and selective surface modifications were incorporated on injection molded polypropylene surfaces. Distinctive lubrication improvement was observed when the surface structure density for the lubricant storage was high, and the release of the lubricant was controlled by the interaction with the selectively modified surfaces.

  4. Progress on nuclear modifications of structure functions

    CERN Document Server

    Kumano, S

    2016-01-01

    We report progress on nuclear structure functions, especially on their nuclear modifications and a new tensor structure function for the deuteron. To understand nuclear structure functions is an important step toward describing nuclei and QCD matters from low to high densities and from low to high energies in terms of fundamental quark and gluon degrees of freedom beyond conventional hadron and nuclear physics. It is also practically important for understanding new phenomena in high-energy heavy-ion collisions at RHIC and LHC. Furthermore, since systematic errors of current neutrino-oscillation experiments are dominated by uncertainties of neutrino-nucleus interactions, such studies are valuable for finding new physics beyond current framework. Next, a new tensor-polarized structure function $b_1$ is discussed for the deuteron. There was a measurement by HERMES; however, its data are inconsistent with the conventional convolution estimate based on the standard deuteron model with D-state admixture. This fact ...

  5. Epigenetic Modifications of Major Depressive Disorder.

    Science.gov (United States)

    Saavedra, Kathleen; Molina-Márquez, Ana María; Saavedra, Nicolás; Zambrano, Tomás; Salazar, Luis A

    2016-08-05

    Major depressive disorder (MDD) is a chronic disease whose neurological basis and pathophysiology remain poorly understood. Initially, it was proposed that genetic variations were responsible for the development of this disease. Nevertheless, several studies within the last decade have provided evidence suggesting that environmental factors play an important role in MDD pathophysiology. Alterations in epigenetics mechanism, such as DNA methylation, histone modification and microRNA expression could favor MDD advance in response to stressful experiences and environmental factors. The aim of this review is to describe genetic alterations, and particularly altered epigenetic mechanisms, that could be determinants for MDD progress, and how these alterations may arise as useful screening, diagnosis and treatment monitoring biomarkers of depressive disorders.

  6. Modification of Food Systems by Ultrasound

    Directory of Open Access Journals (Sweden)

    L. M. Carrillo-Lopez

    2017-01-01

    Full Text Available This review describes the mechanism, operation, and recent potential applications of ultrasound in various food systems, as well as the physical and chemical effects of ultrasound treatments on the conservation and modification of different groups of food. Acoustic energy has been recognized as an emerging technology with great potential for applications in the food industry. The phenomenon of acoustic cavitation, which modifies the physical, chemical, and functional properties of food, can be used to improve existing processes and to develop new ones. The combination of ultrasonic energy with a sanitizing agent can improve the effect of microbial reduction in foods and, thereby, their quality. Finally, it is concluded that the use of ultrasound in food is a very promising area of research; however, more research is still needed before applying this technology in a wider range of industrial sectors.

  7. Cryochemical modification, activity, and toxicity of dioxidine

    Science.gov (United States)

    Vernaya, O. I.; Shabatin, V. P.; Shabatina, T. I.; Khvatov, D. I.; Semenov, A. M.; Yudina, T. P.; Danilov, V. S.

    2017-02-01

    Dioxidine nanoparticles are prepared via cryochemical modification of the pharmacopoeial dioxidine substance. The form of the cryomodified dioxidine is characterized by data from 1H NMR spectroscopy; X-ray diffraction analysis; such thermal analytical methods as TG and DSC; low-temperature argon adsorption; and transmission electron microscopy. It is shown that the cryomodified samples are synthesized in the form of dioxidine nanocrystals 50-300 nm in size, with a crystal structure differing from that of the initial pharmacopoeial substance. The prepared cryomodified dioxidine nanoparticles inhibit the growth of E. coli 52, S. aureus 144, M. cyaneum 98, and B. cereus 9 better than the initial pharmacopoeial substance, and have comparable chronic toxicity.

  8. Pursuing DNA catalysts for protein modification.

    Science.gov (United States)

    Silverman, Scott K

    2015-05-19

    Catalysis is a fundamental chemical concept, and many kinds of catalysts have considerable practical value. Developing entirely new catalysts is an exciting challenge. Rational design and screening have provided many new small-molecule catalysts, and directed evolution has been used to optimize or redefine the function of many protein enzymes. However, these approaches have inherent limitations that prompt the pursuit of different kinds of catalysts using other experimental methods. Nature evolved RNA enzymes, or ribozymes, for key catalytic roles that in modern biology are limited to phosphodiester cleavage/ligation and amide bond formation. Artificial DNA enzymes, or deoxyribozymes, have great promise for a broad range of catalytic activities. They can be identified from unbiased (random) sequence populations as long as the appropriate in vitro selection strategies can be implemented for their identification. Notably, in vitro selection is different in key conceptual and practical ways from rational design, screening, and directed evolution. This Account describes the development by in vitro selection of DNA catalysts for many different kinds of covalent modification reactions of peptide and protein substrates, inspired in part by our earlier work with DNA-catalyzed RNA ligation reactions. In one set of studies, we have sought DNA-catalyzed peptide backbone cleavage, with the long-term goal of artificial DNA-based proteases. We originally anticipated that amide hydrolysis should be readily achieved, but in vitro selection instead surprisingly led to deoxyribozymes for DNA phosphodiester hydrolysis; this was unexpected because uncatalyzed amide bond hydrolysis is 10(5)-fold faster. After developing a suitable selection approach that actively avoids DNA hydrolysis, we were able to identify deoxyribozymes for hydrolysis of esters and aromatic amides (anilides). Aliphatic amide cleavage remains an ongoing focus, including via inclusion of chemically modified DNA

  9. Enzymatic Modification of Plant Cell Wall Polysaccharides

    DEFF Research Database (Denmark)

    Øbro, Jens; Hayashi, Takahisa; Mikkelsen, Jørn Dalgaard

    2011-01-01

    for sustainable processes that replace chemical treatments with white biotechnology. Plants can contribute significantly to this sustainable process by producing plant or microbialenzymes in planta that are necessary for plant cell wall modification or total degradation. This will give rise to superior food......Plant cell walls are intricate structures with remarkable properties, widely used in almost every aspect of our life. Cell walls consist largely of complex polysaccharides and there is often a need for chemical and biochemical processing before industrial use. There is an increasing demand...... fibres, hydrocolloids, paper,textile, animal feeds or biofuels. Classical microbial-based fermentation systems could in the future face serious competition from plant-based expression systems for enzyme production. Plant expressed enzymes can either be targeted to specific cellular compartments...

  10. Enzymatic Modification of Plant Cell Wall Polysaccharides

    DEFF Research Database (Denmark)

    Øbro, Jens; Hayashi, Takahisa; Mikkelsen, Jørn Dalgaard

    2011-01-01

    fibres, hydrocolloids, paper,textile, animal feeds or biofuels. Classical microbial-based fermentation systems could in the future face serious competition from plant-based expression systems for enzyme production. Plant expressed enzymes can either be targeted to specific cellular compartments......Plant cell walls are intricate structures with remarkable properties, widely used in almost every aspect of our life. Cell walls consist largely of complex polysaccharides and there is often a need for chemical and biochemical processing before industrial use. There is an increasing demand...... for sustainable processes that replace chemical treatments with white biotechnology. Plants can contribute significantly to this sustainable process by producing plant or microbialenzymes in planta that are necessary for plant cell wall modification or total degradation. This will give rise to superior food...

  11. Biomolecular Modification of Inorganic Crystal Growth

    Energy Technology Data Exchange (ETDEWEB)

    De Yoreo, J J

    2007-04-27

    The fascinating shapes and hierarchical designs of biomineralized structures are an inspiration to materials scientists because of the potential they suggest for biomolecular control over materials synthesis. Conversely, the failure to prevent or limit tissue mineralization in the vascular, skeletal, and urinary systems is a common source of disease. Understanding the mechanisms by which organisms direct or limit crystallization has long been a central challenge to the biomineralization community. One prevailing view is that mineral-associated macromolecules are responsible for either inhibiting crystallization or initiating and stabilizing non-equilibrium crystal polymorphs and morphologies through interactions between anionic moieties and cations in solution or at mineralizing surfaces. In particular, biomolecules that present carboxyl groups to the growing crystal have been implicated as primary modulators of growth. Here we review the results from a combination of in situ atomic force microscopy (AFM) and molecular modeling (MM) studies to investigate the effect of specific interactions between carboxylate-rich biomolecules and atomic steps on crystal surfaces during the growth of carbonates, oxalates and phosphates of calcium. Specifically, we how the growth kinetics and morphology depend on the concentration of additives that include citrate, simple amino acids, synthetic Asp-rich polypeptides, and naturally occurring Asp-rich proteins found in both functional and pathological mineral tissues. The results reveal a consistent picture of shape modification in which stereochemical matching of modifiers to specific atomic steps drives shape modification. Inhibition and other changes in growth kinetics are shown to be due to a range of mechanisms that depend on chemistry and molecular size. Some effects are well described by classic crystal growth theories, but others, such as step acceleration due to peptide charge and hydrophylicity, were previously unrealized

  12. Seismic force modification factor for ductile structures

    Institute of Scientific and Technical Information of China (English)

    TONG Gen-shu; HUANG Jin-qiao

    2005-01-01

    The earthquake forces used in design codes of buildings should be theoretically determinable. This work examines the seismic force modification factor R based on elastic-plastic time-history earthquake analysis of SDOF systems, wherein the hysteresis models are elastic-perfectly-plastic (EPP), elastic-linearly-hardening (ELH), shear-slipped and bilinear-elastic. The latter two models are analysed for separating the effect of the ductility and the energy-dissipating capacity. Three-hundred eighty-eight earthquake records from different site conditions are used in analysis. The ductility is taken to be 2, 3, 4, 5 and 6, with the damping ratio being 0.02, 0.035 and 0.05 respectively. The post-yield stiffness ratios 0.0, 0.1 and 0.2 are used in the analysis. The R spectra are standardized by the characteristic period of the earthquake records, which leads to a much smaller scatter in averaged numerical results. It was found that the most important factor determining R is the ductility. R increases more than linearly with ductility. The energy-dissipating capacity, damping and the post-yield stiffness are the less important factors. The energy dissipating capacity is important only for structures with short period and moderate period (0.3≤T/Tg<5.0). For EPP and ELH models, R for 0.05 damping is 10% to 15% smaller than for 0.02 damping. For EPP and ELH models, greater post-yield stiffness leads to greater R, but the influence of post-yield stiffness is obvious only when the post-yield stiffness is less than 10% of the initial stiffness. By means of statistical regression analysis the relation of the seismic force modification factor R with the natural period of the system and ductility for EPP and ELH models were established for each site and soil condition.

  13. Bivalent histone modifications during tooth development

    Institute of Scientific and Technical Information of China (English)

    Li-Wei Zheng; Bin-Peng Zhang; Ruo-Shi Xu; Xin Xu; Ling Ye; Xue-Dong Zhou

    2014-01-01

    Histone methylation is one of the most widely studied post-transcriptional modifications. It is thought to be an important epigenetic event that is closely associated with cell fate determination and differentiation. To explore the spatiotemporal expression of histone H3 lysine 4 trimethylation (H3K4me3) and histone H3 lysine 27 trimethylation (H3K27me3) epigenetic marks and methylation or demethylation transferases in tooth organ development, we measured the expression of SET7, EZH2, KDM5B and JMJD3 via immunohistochemistry and quantitative polymerase chain reaction (qPCR) analysis in the first molar of BALB/c mice embryos at E13.5, E15.5, E17.5, P0 and P3, respectively. We also measured the expression of H3K4me3 and H3K27me3 with immunofluorescence staining. During murine tooth germ development, methylation or demethylation transferases were expressed in a spatial–temporal manner. The bivalent modification characterized by H3K4me3 and H3K27me3 can be found during the tooth germ development, as shown by immunofluorescence. The expression of SET7, EZH2 as methylation transferases and KDM5B and JMJD3 as demethylation transferases indicated accordingly with the expression of H3K4me3 and H3K27me3 respectively to some extent. The bivalent histone may play a critical role in tooth organ development via the regulation of cell differentiation.

  14. One-pot chemical synthesis of small ubiquitin-like modifier protein-peptide conjugates using bis(2-sulfanylethyl)amido peptide latent thioester surrogates.

    Science.gov (United States)

    Boll, Emmanuelle; Drobecq, Hervé; Ollivier, Nathalie; Blanpain, Annick; Raibaut, Laurent; Desmet, Rémi; Vicogne, Jérôme; Melnyk, Oleg

    2015-02-01

    Small ubiquitin-like modifier (SUMO) post-translational modification (PTM) of proteins has a crucial role in the regulation of important cellular processes. This protocol describes the chemical synthesis of functional SUMO-peptide conjugates. The two crucial stages of this protocol are the solid-phase synthesis of peptide segments derivatized by thioester or bis(2-sulfanylethyl)amido (SEA) latent thioester functionalities and the one-pot assembly of the SUMO-peptide conjugate by a sequential native chemical ligation (NCL)/SEA native peptide ligation reaction sequence. This protocol also enables the isolation of a SUMO SEA latent thioester, which can be attached to a target peptide or protein in a subsequent step. It is compatible with 9-fluorenylmethoxycarbonyl (Fmoc) chemistry, and it gives access to homogeneous, reversible and functional SUMO conjugates that are not easily produced using living systems. The synthesis of SUMO-peptide conjugates on a milligram scale takes 20 working days.

  15. Monopoles and Modifications of Bundles over Elliptic Curves

    Directory of Open Access Journals (Sweden)

    Andrey M. Levin

    2009-06-01

    Full Text Available Modifications of bundles over complex curves is an operation that allows one to construct a new bundle from a given one. Modifications can change a topological type of bundle. We describe the topological type in terms of the characteristic classes of the bundle. Being applied to the Higgs bundles modifications establish an equivalence between different classical integrable systems. Following Kapustin and Witten we define the modifications in terms of monopole solutions of the Bogomolny equation. We find the Dirac monopole solution in the case R × (elliptic curve. This solution is a three-dimensional generalization of the Kronecker series. We give two representations for this solution and derive a functional equation for it generalizing the Kronecker results. We use it to define Abelian modifications for bundles of arbitrary rank. We also describe non-Abelian modifications in terms of theta-functions with characteristic.

  16. Profile modification to minimize spur gear dynamic loading

    Science.gov (United States)

    Lin, H. H.; Townsend, D. P.; Oswald, F. B.

    1989-01-01

    An analytical computer simulation program for dynamic modeling of low-contact-ratio spur gear systems is presented. The procedure computes the static transmission error of the gears operating under load and uses a fast Fourier transform to generate the frequency spectrum of the static transmission error at various tooth profile modifications. The dynamic loading response of an unmodified (perfect involute) gear pair was compared with that of gears with various profile modifications. Correlations were found between various profile modifications and the resulting dynamic loads. An effective error, obtained from frequency domain analysis of the static transmission error of the gears, gave a very good indication of the optimum profile modification to reduce gear dynamic loading. Design curves generated by dynamic simulation at various profile modifications are given for gear systems operated at various loads. Optimum profile modifications can be determined from these design curves for improved gear design.

  17. On the genetic modification of psychology, personality, and behavior.

    Science.gov (United States)

    Neitzke, Alex B

    2012-12-01

    I argue that the use of heritable modifications for psychology, personality, and behavior should be limited to the reversal or prevention of relatively unambiguous instances of pathology or likely harm (e.g. sociopathy). Most of the likely modifications of psychological personality would not be of this nature, however, and parents therefore should not have the freedom to make such modifications to future children. I argue by examining the viewpoints of both the individual and society. For individuals, modifications would interfere with their capacity for self-determination in a way that undermines the very concept of self-determination. I argue that modification of psychology and personality is unlike present parenting in morally significant ways. For society, modification offers a medium for power to manipulate the makeup of persons and populations, possibly causing biological harm to the species and altering our conceptions of social responsibility.

  18. Land cover modification geoindicator applied in a tropical coastal environment

    OpenAIRE

    Palacio-Aponte, Gerardo

    2014-01-01

    Environmental changes due to natural processes and anthropic modifications can be characterized by the degree of land cover modification and its environmental implications over time. The main goal of the present study was to propose and apply a land cover modification geoindicator in order to assess the environmental condition of the territory per landscape units. It was designed to interpret diffuse information and transform it into a synthetic indicator that will be useful for environmental...

  19. Effect Modification and Interaction Terms: It Takes Two to Tango.

    Science.gov (United States)

    Jupiter, Daniel C

    2016-01-01

    In this Investigators' Corner I look more deeply into the previously discussed phenomenon of effect modification. I revisit an explanation and examples of the phenomenon and then examine how to account for it statistically. Specifically, I show, in detail, how to write a regression equation that includes interaction terms that account for the effect modification. Finally, I look at interpretation of regression coefficients both with and without the presence of effect modification, and the associated interaction terms.

  20. Modification of JRR-4 based on safety evaluation

    Energy Technology Data Exchange (ETDEWEB)

    Izumo, Hironobu; Nakajima, Teruo; Funayama, Yoshiro [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment

    1998-10-01

    Since the first criticality was achieved on January 28, 1965, JRR-4 has been operated safely until on January 12, 1996. The modification of JRR-4 was planned according to the framework of reduced enrichment on research reactor program. The modification was designed based on the several national safety guides. JRR-4 has some modifications of facilities to satisfy the guides and guides criteria. (author)

  1. Assays for investigating deSUMOylation enzymes.

    Science.gov (United States)

    Madu, Ikenna G; Chen, Yuan

    2012-07-01

    Post-translational modifications by the SUMO (Small Ubiquitin-like MOdifier) family of proteins are recently discovered essential regulatory mechanisms. All SUMO proteins are synthesized as larger precursors that are matured by SUMO-specific proteases, known as SENPs, which remove several C-terminal amino acids of SUMO to expose the Gly-Gly motif. SENPs also remove SUMO modifications from target proteins, making this modification highly dynamic. At least six deSUMOylation enzymes, all of which are encoded by essential genes, have been identified in mammals. SENP1 has been shown to play an important role in the development of prostate cancer and in angiogenesis. This unit describes and discusses methods for characterizing the deSUMOylation enzymes. These assays enable the identification of inhibitors of these enzymes and investigation of their mechanism of inhibition in order to develop research tools and future therapeutics.

  2. Assays for investigating deSUMOylation enzymes (ms# CP-11-0260)

    Science.gov (United States)

    Madu, Ikenna G.; Chen, Yuan

    2012-01-01

    Post-translational modifications by the SUMO (Small Ubiquitin-like MOdifier) family of proteins are recently discovered essential regulatory mechanisms. All SUMO proteins are synthesized as larger precursors that are matured by SUMO-specific proteases, known as SENPs, which remove several C-terminal amino acids of SUMO to expose the Gly-Gly motif. SENPs also remove SUMO modifications from target proteins, making this modification highly dynamic. At least six deSUMOylation enzymes, all of which are encoded by essential genes, have been identified in mammals. SENP1 has been shown to play an important role in the development of prostate cancer and in angiogenesis. This unit describes and discusses methods for characterizing the deSUMOylation enzymes. These assays enable the identification of inhibitors of these enzymes and investigate their mechanism of inhibition in order to develop research tools and future therapeutics. PMID:22870856

  3. Genetic modification of preimplantation embryos: toward adequate human research policies.

    Science.gov (United States)

    Dresser, Rebecca

    2004-01-01

    Citing advances in transgenic animal research and setbacks in human trials of somatic cell genetic interventions, some scientists and others want to begin planning for research involving the genetic modification of human embryos. Because this form of genetic modification could affect later-born children and their offspring, the protection of human subjects should be a priority in decisions about whether to proceed with such research. Yet because of gaps in existing federal policies, embryo modification proposals might not receive adequate scientific and ethical scrutiny. This article describes current policy shortcomings and recommends policy actions designed to ensure that the investigational genetic modification of embryos meets accepted standards for research on human subjects.

  4. Single-nucleosome mapping of histone modifications in S. cerevisiae.

    Directory of Open Access Journals (Sweden)

    Chih Long Liu

    2005-10-01

    Full Text Available Covalent modification of histone proteins plays a role in virtually every process on eukaryotic DNA, from transcription to DNA repair. Many different residues can be covalently modified, and it has been suggested that these modifications occur in a great number of independent, meaningful combinations. Published low-resolution microarray studies on the combinatorial complexity of histone modification patterns suffer from confounding effects caused by the averaging of modification levels over multiple nucleosomes. To overcome this problem, we used a high-resolution tiled microarray with single-nucleosome resolution to investigate the occurrence of combinations of 12 histone modifications on thousands of nucleosomes in actively growing S. cerevisiae. We found that histone modifications do not occur independently; there are roughly two groups of co-occurring modifications. One group of lysine acetylations shows a sharply defined domain of two hypo-acetylated nucleosomes, adjacent to the transcriptional start site, whose occurrence does not correlate with transcription levels. The other group consists of modifications occurring in gradients through the coding regions of genes in a pattern associated with transcription. We found no evidence for a deterministic code of many discrete states, but instead we saw blended, continuous patterns that distinguish nucleosomes at one location (e.g., promoter nucleosomes from those at another location (e.g., over the 3' ends of coding regions. These results are consistent with the idea of a simple, redundant histone code, in which multiple modifications share the same role.

  5. Aircraft Modifications: Assessing the Current State of Air Force Aircraft Modifications and the Implications for Future Military Capability

    Science.gov (United States)

    2007-01-25

    Modifications, analysis suggests that installation and procurement schedules are codependent due to an Air Force requirement to install a modification kit in...simultaneously and codependently determined. Further exploration of this requirement is presented in Chapter 5. Schedule estimates is combined with the cost...install a modification kit in the same year that it is procured will constrain one or both of these phases by making them codependent . In accordance with

  6. Fabrication and Modification of Nanoporous Silicon Particles

    Science.gov (United States)

    Ferrari, Mauro; Liu, Xuewu

    2010-01-01

    Silicon-based nanoporous particles as biodegradable drug carriers are advantageous in permeation, controlled release, and targeting. The use of biodegradable nanoporous silicon and silicon dioxide, with proper surface treatments, allows sustained drug release within the target site over a period of days, or even weeks, due to selective surface coating. A variety of surface treatment protocols are available for silicon-based particles to be stabilized, functionalized, or modified as required. Coated polyethylene glycol (PEG) chains showed the effective depression of both plasma protein adsorption and cell attachment to the modified surfaces, as well as the advantage of long circulating. Porous silicon particles are micromachined by lithography. Compared to the synthesis route of the nanomaterials, the advantages include: (1) the capability to make different shapes, not only spherical particles but also square, rectangular, or ellipse cross sections, etc.; (2) the capability for very precise dimension control; (3) the capacity for porosity and pore profile control; and (4) allowance of complex surface modification. The particle patterns as small as 60 nm can be fabricated using the state-of-the-art photolithography. The pores in silicon can be fabricated by exposing the silicon in an HF/ethanol solution and then subjecting the pores to an electrical current. The size and shape of the pores inside silicon can be adjusted by the doping of the silicon, electrical current application, the composition of the electrolyte solution, and etching time. The surface of the silicon particles can be modified by many means to provide targeted delivery and on-site permanence for extended release. Multiple active agents can be co-loaded into the particles. Because the surface modification of particles can be done on wafers before the mechanical release, asymmetrical surface modification is feasible. Starting from silicon wafers, a treatment, such as KOH dipping or reactive ion

  7. Modification of Nanostructures via Laser Processing

    Science.gov (United States)

    Franzel, Louis Avery

    Modification of nanostructures via laser processing is of great interest for a wide range of applications such as aerospace and the storage of nuclear waste. The primary goal of this dissertation is to improve the understanding of nanostructures through two primary routes: the modification of aerogels and pulsed laser ablation in ethanol. A new class of materials, patterned aerogels, was fabricated by photopolymerizing selected regions of homogeneous aerogel monoliths using visible light. The characterization and fabrication of functionally graded, cellular and compositionally anisotropic aerogels and ceramics is discussed. Visible light was utilized due to it's minimal absorption and scattering by organic molecules and oxide nanoparticles within wet gels. This allowed for the fabrication of deeply penetrating, well resolved patterns. Similarly, nanoporous monoliths with a typical aerogel core and a mechanically robust exterior ceramic layer were synthesized from silica aerogels cross-linked with polyacrylonitrile. Simple variations of the exposure geometry allowed fabrication of a wide variety of anisotropic materials without requiring layering or bonding. Nanoparticle solutions were prepared by laser ablation of metal foils (Fe and Mo) in ethanol. Ablation of Fe generated Fe3O4 and Fe3C nanoparticles which were superparamagnetic with a saturation magnetization Ms = 124 emu/g. Zero field cooled (ZFC) measurements collected at an applied field of 50 Oe displayed a maximum magnetic susceptibility at 120 K with a broad distribution. Field cooled (FC) measurements showed a thermal hysteresis indicative of temperature dependent magnetic viscosity. Pulsed laser ablation of a Mo foil in ethanol generated inhomogeneous nanoparticles where Mo and MoC coexisted within the same aggregate. Formation of these unique nanoparticles is likely due to phase separation that occurs when a high temperature carbide phase cools after the laser pulse terminates. Similarly, magnetic

  8. Delivery Ring Lattice Modifications for Transitionless Deceleration

    Energy Technology Data Exchange (ETDEWEB)

    Johnstone, J. A. [Fermilab; Syphers, M. J. [Northern Illinois U.

    2016-10-09

    A portion of the remnant Tevatron program infrastruc- ture at Fermilab is being reconfigured to be used for the generation and delivery of proton and muon beams for new high-precision particle physics experiments. With the 8 GeV Booster as its primary source, the Mu2e exper- iment will receive 8.9 GeV/c bunched beam on target, after being stored and slow spilled from the Delivery Ring (DR) -- a refurbished debuncher ring from Tevatron anti- proton production. For the Muon g-2 experiment, the DR will be tuned for 3.1 GeV/c to capture muons off of a target before sending them to this experiment's Storage Ring. The apertures in the beam transport systems are optimized for the large muon beams of this lower-energy experiment. In order to provide further flexibility in the operation of the DR for future possible low-energy, high- intensity particle physics experiments (REDTOP[1], for example) and detector development, investigations are underway into the feasibility of decelerating beams from its maximum kinetic energy of 8 GeV level to lower en- ergies, down to 1-2 GeV. In this paper we look at possi- ble lattice modifications to the DR to avoid a transition crossing during the deceleration process. Hardware re- quirements and other operational implications of this scheme will also be discussed.

  9. Cysteine Modifications in the Pathogenesis of ALS

    Science.gov (United States)

    Valle, Cristiana; Carrì, Maria Teresa

    2017-01-01

    Several proteins are found misfolded and aggregated in sporadic and genetic forms of amyotrophic lateral sclerosis (ALS). These include superoxide dismutase (SOD1), transactive response DNA-binding protein (TDP-43), fused in sarcoma/translocated in liposarcoma protein (FUS/TLS), p62, vasolin-containing protein (VCP), Ubiquilin-2 and dipeptide repeats produced by unconventional RAN-translation of the GGGGCC expansion in C9ORF72. Up to date, functional studies have not yet revealed a common mechanism for the formation of such diverse protein inclusions. Consolidated studies have demonstrated a fundamental role of cysteine residues in the aggregation process of SOD1 and TDP43, but disturbance of protein thiols homeostatic factors such as protein disulfide isomerases (PDI), glutathione, cysteine oxidation or palmitoylation might contribute to a general aberration of cysteine residues proteostasis in ALS. In this article we review the evidence that cysteine modifications may have a central role in many, if not all, forms of this disease. PMID:28167899

  10. Mobilome and genetic modification of bifidobacteria.

    Science.gov (United States)

    Guglielmetti, S; Mayo, B; Álvarez-Martín, P

    2013-06-01

    Until recently, proper development of molecular studies in Bifidobacterium species has been hampered by growth difficulties, because of their exigent nutritive requirements, oxygen sensitivity and lack of efficient genetic tools. These studies, however, are critical to uncover the cross-talk between bifidobacteria and their hosts' cells and to prove unequivocally the supposed beneficial effects provided through the endogenous bifidobacterial populations or after ingestion as probiotics. The genome sequencing projects of different bifidobacterial strains have provided a wealth of genetic data that will be of much help in deciphering the molecular basis of the physiological properties of bifidobacteria. To this end, the purposeful development of stable cloning and expression vectors based on robust replicons - either from temperate phages or resident plasmids - is still needed. This review addresses the current knowledge on the mobile genetic elements of bifidobacteria (prophages, plasmids and transposons) and summarises the different types of vectors already available, together with the transformation procedures for introducing DNA into the cells. It also covers recent molecular studies performed with such vectors and incipient results on the genetic modification of these organisms, establishing the basis that would allow the use of bifidobacteria for future biotechnological applications.

  11. Surface modification of polypropylene based particle foams

    Science.gov (United States)

    Schreier, P.; Trassl, C.; Altstädt, V.

    2014-05-01

    This paper deals with the modification of the surface properties of expanded polypropylene (EPP). EPP is a semi-hard to soft elastic thermoplastic foam. The characteristic surface of EPP shows process-related steam nozzle imprints and gussets. Therefore EPP does not satisfy the quality requirements for visible automotive applications. In order to meet these demands, plastic surfaces are usually enhanced with functional or decorative coatings, e.g. textiles, plastic films or paint. The coating of plastics with low surface energies such as PP often leads to adhesion problems by reason of the missing polar and functional groups. This paper gives an evaluation of activation and pre-treatment methods of EPP, with the aim to identify the most suitable pre-treatment method. For this purpose five typical surface treatment methods - flame treatment, corona, fluorination, atmospheric and low-pressure plasma - were performed on EPP samples. As a comparison criterion the maximum increase in the adhesion force between a polyurethane-based coating and the modified EPP substrate was selected. Moreover the influence of the selected pre-treatment method on the increase in the total surface energy and its polar component was investigated by the drop shape analysis method. The results showed that the contact angle measurement is a suitable method to determine the polar and disperse fractions of the surface tension of EPP. Furthermore, all performed methods increased the adhesion of EPP.

  12. Progress on nuclear modifications of structure functions

    Directory of Open Access Journals (Sweden)

    Kumano S.

    2016-01-01

    Full Text Available We report progress on nuclear structure functions, especially on their nuclear modifications and a new tensor structure function for the deuteron. To understand nuclear structure functions is an important step toward describing nuclei and QCD matters from low to high densities and from low to high energies in terms of fundamental quark and gluon degrees of freedom beyond conventional hadron and nuclear physics. It is also practically important for understanding new phenomena in high-energy heavy-ion collisions at RHIC and LHC. Furthermore, since systematic errors of current neutrinooscillation experiments are dominated by uncertainties of neutrino-nucleus interactions, such studies are valuable for finding new physics beyond current framework. Next, a new tensor-polarized structure function b1 is discussed for the deuteron. There was a measurement by HERMES; however, its data are inconsistent with the conventional convolution estimate based on the standard deuteron model with D-state admixture. This fact suggests that a new hadronic phenomenon should exist in the tensor-polarized deuteron at high energies, and it will be experimentally investigated at JLab from the end of 2010’s.

  13. Does Memory Modification Threaten Our Authenticity?

    Science.gov (United States)

    Erler, Alexandre

    2011-11-01

    One objection to enhancement technologies is that they might lead us to live inauthentic lives. Memory modification technologies (MMTs) raise this worry in a particularly acute manner. In this paper I describe four scenarios where the use of MMTs might be said to lead to an inauthentic life. I then undertake to justify that judgment. I review the main existing accounts of authenticity, and present my own version of what I call a "true self" account (intended as a complement, rather than a substitute, to existing accounts). I briefly describe current and prospective MMTs, distinguishing between memory enhancement and memory editing. Moving then to an assessment of the initial scenarios in the light of the accounts previously described, I argue that memory enhancement does not, by its very nature, raise serious concerns about authenticity. The main threat to authenticity posed by MMTs comes, I suggest, from memory editing. Rejecting as inadequate the worries about identity raised by the President's Council on Bioethics in Beyond Therapy, I argue instead that memory editing can cause us to live an inauthentic life in two main ways: first, by threatening its truthfulness, and secondly, by interfering with our disposition to respond in certain ways to some past events, when we have reasons to respond in such ways. This consideration allows us to justify the charge of inauthenticity in cases where existing accounts fail. It also gives us a significant moral reason not to use MMTs in ways that would lead to such an outcome.

  14. IMMOBILISATION OF HUMIC SUBSTANCES USING PLASMA MODIFICATION

    Directory of Open Access Journals (Sweden)

    Pavlína Hájková

    2015-04-01

    Full Text Available This paper presents a study of the immobilization of humic substances (HSs on a polypropylene (PP nonwoven fabric. In order to attach the HSs, the PP nonwoven fabric was modified in a volume of nonthermal atmospheric pressure dielectric barrier discharge (DBD under defined conditions. An unmodified PP nonwoven fabric was used as a reference sample. The modified and unmodified samples were both dipped in an aqueous solution of potassium humate, and then the samples were washed in water and the amount of HSs attached to the PP fabric was monitored. An aqueous solution of cadmium salts was filtered through the treated fabric, the content of Cd2+ in the solution was monitored using ICP-OES analysis, and the Cd2+ sorbed on the fabric was proved by SEM/EDS analysis. The efficiency of the PP plasma modification was proved by XPS analysis, and the presence and the distribution of the HSs along the fibers was proved by SEM analysis.

  15. Activated Carbon Preparation and Modification for Adsorption

    Science.gov (United States)

    Cao, Yuhe

    Butanol is considered a promising, infrastructure-compatible biofuel. Butanol has a higher energy content than ethanol and can be used in conventional gas engines without modifications. Unfortunately, the fermentation pathway for butanol production is restricted by its toxicity to the microbial strains used in the process. Butanol is toxic to the microbes, and this can slow fermentation rates and reduce butanol yields. Gas stripping technology can efficiently remove butanol from the fermentation broth as it is produced, thereby decreasing its inhibitory effects. Traditional butanol separation heavily depends on the energy intensive distillation method. One of the main issues in acetone-butanol-ethanol fermentation is that butanol concentrations in the fermentation broth are low, ranging from 1 to 1.2 percent in weight, because of its toxicity to the microorganisms. Therefore distillation of butanol is even worse than distillation of corn ethanol. Even new separation methods, such as solid- extraction methods involve adding substances, such as polymer resin and zeolite or activated carbon, to biobutanol fermentatioon broth did not achieve energy efficient separation of butanol due to low adsorption selectivity and fouling in broth. Gas-stripping - condensation is another new butanol recovery method, however, the butanol in gas-stripping stream is too low to be condensed without using expensive and energy intensive liquid nitrogen. Adsorption can then be used to recover butanol from the vapor phase. Activated carbon (AC) samples and zeolite were investigated for their butanol vapor adsorption capacities. Commercial activated carbon was modified via hydrothermal H2O2 treatment, and the specific surface area and oxygen-containing functional groups of activated carbon were tested before and after treatment. Hydrothermal H2O 2 modification increased the surface oxygen content, Brunauer-Emmett-Teller surface area, micropore volume, and total pore volume of active carbon

  16. Influence of behavior modification on overactive bladder.

    Science.gov (United States)

    Burgio, Kathryn L

    2002-11-01

    Behavioral interventions have been used for decades to treat urge incontinence and other symptoms of overactive bladder. Perhaps the earliest form of treatment was the bladder drill, an intensive intervention designed to increase the interval between voids to establish a normal frequency of urination and normalization of bladder function. Bladder training is a modification of bladder drill that is conducted more gradually on an outpatient basis and has resulted in significant reduction of incontinence in older, community-dwelling women. Multicomponent behavioral training is another form of behavioral treatment that includes pelvic floor muscle training and exercise. This intervention focuses less on voiding habits and more on altering the physiologic responses of the bladder and pelvic floor muscles. Using biofeedback or other teaching methods, patients learn strategies to inhibit bladder contraction using pelvic floor muscle contraction and other urge suppression strategies. Although behavioral and drug therapies are known to be highly effective for reducing urge incontinence, few patients are cured with either treatment alone. Thus, future research should explore ways to enhance the effectiveness of these conservative therapies. Although the mechanisms by which behavioral treatments work have not been established, there is some evidence that behavioral and drug interventions may operate by different mechanisms, suggesting that they may have additive effects and that combining them may result in better outcomes. Future research needs to examine the mechanisms by which these therapies reduce incontinence and whether combining behavioral and drug treatment will result in better outcomes than either therapy alone.

  17. Crystal modifications and dissolution rate of piroxicam.

    Science.gov (United States)

    Lyn, Lim Yee; Sze, Huan Wen; Rajendran, Adhiyaman; Adinarayana, Gorajana; Dua, Kamal; Garg, Sanjay

    2011-12-01

    Piroxicam is a nonsteroidal anti-inflammatory drug with low aqueous solubility which exhibits polymorphism. The present study was carried out to develop polymorphs of piroxicam with enhanced solubility and dissolution rate by the crystal modification technique using different solvent mixtures prepared with PEG 4000 and PVP K30. Physicochemical characteristics of the modified crystal forms of piroxicam were investigated by X-ray powder diffractometry, FT-IR spectrophotometry and differential scanning calorimetry. Dissolution and solubility profiles of each modified crystal form were studied and compared with pure piroxicam. Solvent evaporation method (method I) produced both needle and cubic shaped crystals. Slow crystallization from ethanol with addition of PEG 4000 or PVP K30 at room temperature (method II) produced cubic crystal forms. Needle forms produced by method I improved dissolution but not solubility. Cubic crystals produced by method I had a dissolution profile similar to that of untreated piroxicam but showed better solubility than untreated piroxicam. Cubic shaped crystals produced by method II showed improved dissolution, without a significant change in solubility. Based on the XRPD results, modified piroxicam crystals obtained by method I from acetone/benzene were cube shaped, which correlates well with the FTIR spectrum; modified needle forms obtained from ethanol/methanol and ethanol/acetone showed a slight shift of FTIR peak that may be attributed to differences in the internal structure or conformation.

  18. Modification of cellulose for high glucose generation.

    Science.gov (United States)

    Jiang, Xue; Gu, Jian; Tian, Xiuzhi; Li, Yali; Huang, Dan

    2012-01-01

    The influence of introduction of cyanuric chloride on glucose's yield (Y) in acid-catalyzed hydrolysis of microcrystalline cellulose (MCC) has been studied. The content of cyanuric chloride (C) in modified MCCs was determined by X-ray photoelectric spectroscopy. The chemical structures of modified MCCs were analyzed by Fourier transformation-infrared spectroscopy and cross polarization/magic angle spinning (13)C nuclear magnetic resonance. Crystal index (CI) and the ratio (R) representing the sum of content of (1 ̅10) and (110) to that of (200) were calculated based on diffraction intensity in wide angle X-ray diffraction (WAXD). Hydrolysis experiment and WAXD show that Y, CI and R vary with C. The modified MCC containing 3.9 mol% of cyanuric chloride has the highest Y, the highest R and the lowest CI. Variations of CI and R show that the chemical modification changed the proportion of crystal/amorphous and crystal planes, both of which influence glucose's generation in hydrolysis of cellulose.

  19. Wind Field Modifications in Habitable Urban Areas

    Directory of Open Access Journals (Sweden)

    Seemi Ahmed

    2012-12-01

    Full Text Available This paper discusses different criteria for the assessment of wind field environmentsin urban areas and how they relate to field observations. The importance of the inclusion of wind environment studies in the planning process is also discussed. The increasing influence of the built environment on wind speed and direction makes any forecast for heights below 50 meter very hazardous1. This increase in the areas with more built form where the roughness is extremely high. It is not always possible to make a quantitative forecast of wind speed and direction in urban environment. Examples are provided to illustrate how development controls can be designed to ensure that pedestrian amenity is not compromised by new development while at the same time not become a burden to innovative design approaches or good design practice due to wind modification. The paper concludes with a number of case studies that provide examples of how innovative techniques for mitigation of adverse wind environments can achieve the desired level of pedestrian amenity without having to compromise with the architectural design intent.

  20. Electric Field Induced Surface Modification of Au

    Energy Technology Data Exchange (ETDEWEB)

    Erchak, A.A.; Franklin, G.F.; Houston, J.E.; Mayer, T.M.; Michalske, T.A.

    1999-02-15

    We discuss the role of localized high electric fields in the modification of Au surfaces with a W probe using the Interfacial Force Microscope. Upon bringing a probe close to a Au surface, we measure both the interfacial force and the field emission current as a function of separation with a constant potential of 100 V between tip and sample. The current initially increases exponentially as the separation decreases. However, at a distance of less than {approximately} 500{angstrom} the current rises sharply as the surface begins to distort and rapidly close the gap. Retraction of the tip before contact is made reveals the formation of a mound on the surface. We propose a simple model, in which the localized high electric field under the tip assists the production of mobile Au adatoms by detachment from surface steps, and a radial field gradient causes a net flux of atoms toward the tip by surface diffusion. These processes give rise to an unstable surface deformation which, if left unchecked, results in a destructive mechanical contact. We discuss our findings with respect to earlier work using voltage pulses in the STM as a means of nanofabrication.

  1. Attentional bias modification encourages healthy eating.

    Science.gov (United States)

    Kakoschke, Naomi; Kemps, Eva; Tiggemann, Marika

    2014-01-01

    The continual exposure to unhealthy food cues in the environment encourages poor dietary habits, in particular consuming too much fat and sugar, and not enough fruit and vegetables. According to Berridge's (2009) model of food reward, unhealthy eating is a behavioural response to biased attentional processing. The present study used an established attentional bias modification paradigm to discourage the consumption of unhealthy food and instead promote healthy eating. Participants were 146 undergraduate women who were randomly assigned to two groups: one was trained to direct their attention toward pictures of healthy food ('attend healthy' group) and the other toward unhealthy food ('attend unhealthy' group). It was found that participants trained to attend to healthy food cues demonstrated an increased attentional bias for such cues and ate relatively more of the healthy than unhealthy snacks compared to the 'attend unhealthy' group. Theoretically, the results support the postulated link between biased attentional processing and consumption (Berridge, 2009). At a practical level, they offer potential scope for interventions that focus on eating well. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Heavy Flavored Jet Modification at CMS

    Science.gov (United States)

    Jung, Kurt

    2016-12-01

    The energy loss of jets in heavy-ion collisions is expected to depend on the flavor of the fragmenting parton. Thus, measurements of jet quenching as a function of flavor place powerful constraints on the thermodynamical and transport properties of the hot and dense medium. Specifically, pPb measurements provide crucial insights into the behavior of the cold nuclear matter effect, which is required to fully understand the hot and dense medium effects on jets in PbPb collisions. In these proceedings, we present the heavy flavor jet spectra and measurements of the nuclear modification factors of b jets in both PbPb and pPb as a function of transverse momentum and pseudorapidity, at √{sNN} = 2.76 and 5.02 TeV, respectively. In addition, we present the first ever measurement of charm-tagged jets in a heavy-ion environment, including cross-sections and comparisons to PYTHIA in both pPb and pp.

  3. Surface modification using ionic liquid ion beams

    Energy Technology Data Exchange (ETDEWEB)

    Takaoka, Gikan H.; Hamaguchi, Takuya; Takeuchi, Mitsuaki; Ryuto, Hiromichi

    2014-12-15

    We developed an ionic liquid (IL) ion source using 1-butyl-3-methylimidazolium hexafluorophosphate (BMIM-PF{sub 6}) and produced IL ion beams by applying a high electric field between the tip and the extractor. Time-of-flight measurements showed that small cluster and fragment ions were contained in the positive and negative ion beams. The positive and negative cluster ions were deposited on Si(1 0 0) substrates. X-ray photoelectron spectroscopy measurements showed that the composition of the deposited layers was similar to that of an IL solvent. This suggests that a cation (A{sup +}) or an anion (B{sup −}) was attached to an IL cluster (AB){sub n}, resulting in the formation of positive cluster ions (AB){sub n}A{sup +} or negative cluster ions (AB){sub n}B{sup −}, respectively. The surfaces of the IL layers deposited on Si(1 0 0) substrates were flat at an atomic level for positive and negative cluster ion irradiation. Moreover, the contact angles of the deposited layers were similar to that of the IL solvent. Thus, surface modification of Si(1 0 0) substrates was successfully demonstrated with BMIM-PF{sub 6} cluster ion beams.

  4. Posttranslational modification of a vanadium nitrogenase.

    Science.gov (United States)

    Heiniger, Erin K; Harwood, Caroline S

    2015-08-01

    In microbes that fix nitrogen, nitrogenase catalyzes the conversion of N2 to ammonia in an ATP-demanding reaction. To help conserve energy some bacteria inhibit nitrogenase activity upon exposure to ammonium. The purple nonsulfur phototrophic bacterium Rhodopseudomonas palustris strain CGA009 can synthesize three functional nitrogenase isoenzymes: a molybdenum nitrogenase, a vanadium nitrogenase, and an iron nitrogenase. Previous studies showed that in some alphaproteobacteria, including R. palustris, molybdenum nitrogenase activity is inhibited by ADP-ribosylation when cells are exposed to ammonium. Some iron nitrogenases are also posttranslationally modified. However, the posttranslational modification of vanadium nitrogenase has not been reported. Here, we investigated the regulation of the alternative nitrogenases of R. palustris and determined that both its vanadium nitrogenase and its iron nitrogenase activities were inhibited and posttranslationally modified when cells are exposed to ammonium. Vanadium nitrogenase is not found in all strains of R. palustris, suggesting that it may have been acquired by horizontal gene transfer. Also, phylogenetic analyses of the three nitrogenases suggest that VnfH, the target of ADP-ribosylation, may be the product of a gene duplication of nifH, the molybdenum nitrogenase homolog.

  5. The modification behaviour for Si implanted PET

    Institute of Scientific and Technical Information of China (English)

    吴瑜光; 张通和; 刘安东; 张旭; 周固

    2003-01-01

    Polyethylene terephthalate (PET) has been modified by Si ion implantation with a dose ranging from 1 × 1016 to 2 × 1017 ions /cm2 using a metal vapor vacuum arc(MEVVA)source. The surface morphology was observed by atomic force microscopy (AFM). The change in the microstructure of Si implanted PET was observed with a transmission electron microscope (TEM). It is believed that the change would improve the conductive properties and wear resistance. The electrical properties of PET have been improved via Si ion implantation. The resistivity of implanted PET decreased obviously with an increase in ion dose. When Si ion dose was 2 × 1017 cm?2, the resistivity of PET could be less than 7.9 Ω@m. The surface hardness and modulus increased obviously. The mechanical property of the implanted PET has been modified greatly. The hardness and modulus of Si implanted PET with a dose of 2 × 1017/cm2 are 12.5 and 2.45 times greater than those of pristine PET, respectively. The area of cutting groove for Si implanted PET is narrower and shallower than those of the unimplanted PET. So the wear resistance is greatly raised. In comparison with metal ion implantation, the improvement of mechanical properties is obvious in ion implantation into PET. Si ion beam modification mechanism of PET is discussed.

  6. Child-focused behavioral assessment and modification.

    Science.gov (United States)

    Evans, I M

    1999-12-01

    Argues that behavioral principles have been translated into practice with children too literally and that a more integrative framework is required to guide assessment and treatment. The framework advocated is Staats's (1996) psychological behaviorism. This is a consistently behavioristic, positivist paradigm, using multilevel theory to emphasize the integration of social learning, developmental, and personality principles. Psychological behaviorism thus allows for a much more expansive approach than has typically been the case within child behavior therapy. Given the complexity of this perspective, I selected four broad tenets of the theory and suggested their implications for clinical contexts. The further translation from clinical models to specific clinical practices is quite difficult but may yield more flexible and substitutable practices than do unidimensional treatment outcome studies. Of special importance, the principles demonstrate how children themselves can retain the central focus of child behavioral assessment and modification. Specific practices still need to be constructed according to an understanding of the multiple sources of influence on children as well as the culture of childhood itself.

  7. Modifications of sequential designs in bioequivalence trials.

    Science.gov (United States)

    Zheng, Cheng; Zhao, Lihui; Wang, Jixian

    2015-01-01

    Bioequivalence (BE) studies are designed to show that two formulations of one drug are equivalent and they play an important role in drug development. When in a design stage, it is possible that there is a high degree of uncertainty on variability of the formulations and the actual performance of the test versus reference formulation. Therefore, an interim look may be desirable to stop the study if there is no chance of claiming BE at the end (futility), or claim BE if evidence is sufficient (efficacy), or adjust the sample size. Sequential design approaches specially for BE studies have been proposed previously in publications. We applied modification to the existing methods focusing on simplified multiplicity adjustment and futility stopping. We name our method modified sequential design for BE studies (MSDBE). Simulation results demonstrate comparable performance between MSDBE and the original published methods while MSDBE offers more transparency and better applicability. The R package MSDBE is available at https://sites.google.com/site/modsdbe/.

  8. Proton beam modification of lead white pigments

    Energy Technology Data Exchange (ETDEWEB)

    Beck, L., E-mail: lucile.beck@cea.fr [CEA, DEN, Service de Recherches de Métallurgie Physique, Laboratoire JANNUS, 91191 Gif-sur-Yvette (France); Centre de recherche et de restauration des musées de France (C2RMF), Palais du Louvre – Porte des Lions, 14 quai François Mitterrand, 75001 Paris (France); Gutiérrez, P.C. [Centre de recherche et de restauration des musées de France (C2RMF), Palais du Louvre – Porte des Lions, 14 quai François Mitterrand, 75001 Paris (France); Centro de Micro-Análisis de Materiales (CMAM), Universidad Autónoma de Madrid, Campus de Cantoblanco, E-28049 Madrid (Spain); Miserque, F. [CEA, DEN, DPC/SCCME/LECA, 91191 Gif-sur-Yvette (France); Thomé, L. [Centre de Spectrométrie Nucléaire et de Spectrométrie de Masse (CSNSM), CNRS/IN2P3 et Université Paris-Sud, Bât. 108, 91405 Orsay (France)

    2013-07-15

    Pigments and paint materials are known to be sensitive to particle irradiation. Occasionally, the analysis of paintings by PIXE can induce a slight or dark stain depending on the experimental conditions (beam current, dose, particle energy). In order to understand this discoloration, we have irradiated various types of art white pigments – lead white (hydrocerussite and basic lead sulfate), gypsum, calcite, zinc oxide and titanium oxide – with an external 3 MeV proton micro-beam commonly used for PIXE experiments. We have observed various sensitivities depending on the pigment. No visible change occurs for calcite and titanium oxide, whereas lead white pigments are very sensitive. For the majority of the studied compounds, the discoloration is proportional to the beam current and charge. The damage induced by proton beam irradiation in lead white pigments was studied by micro-Raman and XPS spectroscopies. Structural modifications and dehydration were detected. Damage recovery was investigated by thermal treatment and UV-light irradiation. The discoloration disappeared after one week of UV illumination, showing that PIXE experiments could be safely undertaken for pigments and paintings.

  9. Comparison of several innovative bridge cable surface modifications

    DEFF Research Database (Denmark)

    Kleissl, Kenneth; Georgakis, Christos T.

    Over the last two decades, several bridge cable manufacturers have introduced surface modifications on the high-density polyethylene (HDPE) sheathing that is installed for the protection of inner cable strands or wires. The modifications are based on research undertaken predominantly in Europe an...

  10. 19 CFR 210.74 - Modification of reporting requirements.

    Science.gov (United States)

    2010-04-01

    ... 19 Customs Duties 3 2010-04-01 2010-04-01 false Modification of reporting requirements. 210.74 Section 210.74 Customs Duties UNITED STATES INTERNATIONAL TRADE COMMISSION INVESTIGATIONS OF UNFAIR... warrants, announce that a modification of reporting is under consideration and ask for comment, but it...

  11. 12 CFR 725.21 - Modification of agreements.

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false Modification of agreements. 725.21 Section 725.21 Banks and Banking NATIONAL CREDIT UNION ADMINISTRATION REGULATIONS AFFECTING CREDIT UNIONS NATIONAL CREDIT UNION ADMINISTRATION CENTRAL LIQUIDITY FACILITY § 725.21 Modification of agreements....

  12. Surface modification of nano-apatite by grafting organic polymer

    NARCIS (Netherlands)

    Liu, Qing; Wijn, de Joost R.; Groot, de Klaas; Blitterswijk, van Clemens A.

    1998-01-01

    Since surface properties of hydroxyapatite (HA) play an important role in its performance, surface modification of HA has gained much attention from researchers. Silane coupling agents have been the focus of the research. In this study, an effective surface modification method was developed using he

  13. 21 CFR 640.6 - Modifications of Whole Blood.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Modifications of Whole Blood. 640.6 Section 640.6...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Whole Blood § 640.6 Modifications of Whole Blood. Upon approval by the Director, Center for Biologics Evaluation and Research, of a supplement...

  14. 25 CFR 11.1013 - Modification of dispositional order.

    Science.gov (United States)

    2010-04-01

    ... OFFENSES AND LAW AND ORDER CODE Juvenile Offender Procedure § 11.1013 Modification of dispositional order... minor or the minor's parents, guardian or custodian. (c) If the modification involves a change of...'s court shall review the performance of the minor, the minor's parents, guardian or custodian, and...

  15. Posttranslational Modification Biology of Glutamate Receptors and Drug Addiction

    Directory of Open Access Journals (Sweden)

    Li-Min eMao

    2011-03-01

    Full Text Available Posttranslational covalent modifications of glutamate receptors remain a hot topic. Early studies have established that this family of receptors, including almost all ionotropic and metabotropic glutamate receptor subtypes, undergoes active phosphorylation at serine, threonine, or tyrosine residues on their intracellular domains. Recent evidence identifies several glutamate receptor subtypes to be direct substrates for palmitoylation at cysteine residues. Other modifications such as ubiquitination and sumoylation at lysine residues also occur to certain glutamate receptors. These modifications are dynamic and reversible in nature and are regulatable by changing synaptic inputs. The regulated modifications significantly impact the receptor in many ways, including interrelated changes in biochemistry (synthesis, subunit assembling and protein-protein interactions, subcellular redistribution (trafficking, endocytosis, synaptic delivery and clustering, and physiology, usually associated with changes in synaptic plasticity. Glutamate receptors are enriched in the striatum and cooperate closely with dopamine to regulate striatal signaling. Emerging evidence shows that modification processes of striatal glutamate receptors are sensitive to addictive drugs, such as psychostimulants (cocaine and amphetamines. Altered modifications are believed to be directly linked to enduring receptor/synaptic plasticity and drug-seeking. This review summarizes several major types of modifications of glutamate receptors and analyzes the role of these modifications in striatal signaling and in the pathogenesis of psychostimulant addiction.

  16. 21 CFR 640.74 - Modification of Source Plasma.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 7 2010-04-01 2010-04-01 false Modification of Source Plasma. 640.74 Section 640...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Source Plasma § 640.74 Modification of Source Plasma. (a) Upon approval by the Director, Center for Biologics Evaluation and Research, Food and...

  17. Beliefs about Child Support Modification Following Remarriage and Subsequent Childbirth

    Science.gov (United States)

    Hans, Jason D.

    2009-01-01

    Framed by equity theory, fairness beliefs regarding child support modification to account for the financial impact of remarriage and subsequent childbirth were assessed. Based on a random sample of 407 Kentucky residents using a multiple segment factorial vignette approach, modification was supported by 57% of respondents following remarriage, but…

  18. 30 CFR 44.52 - Revocation of modification.

    Science.gov (United States)

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Revocation of modification. 44.52 Section 44.52... of Initial Decision § 44.52 Revocation of modification. (a) Petition for revocation. Any party to a.... (b) Revocation by the Administrator. The appropriate Administrator may propose to revoke...

  19. 31 CFR 501.803 - Amendment, modification, or revocation.

    Science.gov (United States)

    2010-07-01

    ... 31 Money and Finance: Treasury 3 2010-07-01 2010-07-01 false Amendment, modification, or revocation. 501.803 Section 501.803 Money and Finance: Treasury Regulations Relating to Money and Finance... REGULATIONS Procedures § 501.803 Amendment, modification, or revocation. Except as otherwise provided by...

  20. 47 CFR 76.59 - Modification of television markets.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 4 2010-10-01 2010-10-01 false Modification of television markets. 76.59... MULTICHANNEL VIDEO AND CABLE TELEVISION SERVICE Carriage of Television Broadcast Signals § 76.59 Modification of television markets. (a) The Commission, following a written request from a broadcast station or...