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Sample records for adp ribosylating bacterial

  1. The ARTT motif and a unified structural understanding of substraterecognition in ADP ribosylating bacterial toxins and eukaryotic ADPribosyltransferases

    Energy Technology Data Exchange (ETDEWEB)

    Han, S.; Tainer, J.A.

    2001-08-01

    ADP-ribosylation is a widely occurring and biologically critical covalent chemical modification process in pathogenic mechanisms, intracellular signaling systems, DNA repair, and cell division. The reaction is catalyzed by ADP-ribosyltransferases, which transfer the ADP-ribose moiety of NAD to a target protein with nicotinamide release. A family of bacterial toxins and eukaryotic enzymes has been termed the mono-ADP-ribosyltransferases, in distinction to the poly-ADP-ribosyltransferases, which catalyze the addition of multiple ADP-ribose groups to the carboxyl terminus of eukaryotic nucleoproteins. Despite the limited primary sequence homology among the different ADP-ribosyltransferases, a central cleft bearing NAD-binding pocket formed by the two perpendicular b-sheet core has been remarkably conserved between bacterial toxins and eukaryotic mono- and poly-ADP-ribosyltransferases. The majority of bacterial toxins and eukaryotic mono-ADP-ribosyltransferases are characterized by conserved His and catalytic Glu residues. In contrast, Diphtheria toxin, Pseudomonas exotoxin A, and eukaryotic poly-ADP-ribosyltransferases are characterized by conserved Arg and catalytic Glu residues. The NAD-binding core of a binary toxin and a C3-like toxin family identified an ARTT motif (ADP-ribosylating turn-turn motif) that is implicated in substrate specificity and recognition by structural and mutagenic studies. Here we apply structure-based sequence alignment and comparative structural analyses of all known structures of ADP-ribosyltransfeases to suggest that this ARTT motif is functionally important in many ADP-ribosylating enzymes that bear a NAD binding cleft as characterized by conserved Arg and catalytic Glu residues. Overall, structure-based sequence analysis reveals common core structures and conserved active sites of ADP-ribosyltransferases to support similar NAD binding mechanisms but differing mechanisms of target protein binding via sequence variations within the ARTT

  2. An Entamoeba histolytica ADP-ribosyl transferase from the diphtheria toxin family modifies the bacterial elongation factor Tu.

    Science.gov (United States)

    Avila, Eva E; Rodriguez, Orlando I; Marquez, Jaqueline A; Berghuis, Albert M

    2016-06-01

    ADP-ribosyl transferases are enzymes involved in the post-translational modification of proteins; they participate in multiple physiological processes, pathogenesis and host-pathogen interactions. Several reports have characterized the functions of these enzymes in viruses, prokaryotes and higher eukaryotes, but few studies have reported ADP-ribosyl transferases in lower eukaryotes, such as parasites. The locus EHI_155600 from Entamoeba histolytica encodes a hypothetical protein that possesses a domain from the ADP-ribosylation superfamily; this protein belongs to the diphtheria toxin family according to a homology model using poly-ADP-ribosyl polymerase 12 (PARP12 or ARTD12) as a template. The recombinant protein expressed in Escherichia coli exhibited in vitro ADP-ribosylation activity that was dependent on the time and temperature. Unlabeled βNAD(+), but not ADP-ribose, competed in the enzymatic reaction using biotin-βNAD(+) as the ADP-ribose donor. The recombinant enzyme, denominated EhToxin-like, auto-ADP-ribosylated and modified an acceptor from E. coli that was identified by MS/MS as the elongation factor Tu (EF-Tu). To the best of our knowledge, this is the first report to identify an ADP-ribosyl transferase from the diphtheria toxin family in a protozoan parasite. The known toxins from this family (i.e., the diphtheria toxin, the Pseudomonas aeruginosa toxin Exo-A, and Cholix from Vibrio cholerae) modify eukaryotic elongation factor two (eEF-2), whereas the amoeba EhToxin-like modified EF-Tu, which is another elongation factor involved in protein synthesis in bacteria and mitochondria. PMID:27234208

  3. The natural history of ADP-ribosyltransferases and the ADP-ribosylation system.

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    Aravind, L; Zhang, Dapeng; de Souza, Robson F; Anand, Swadha; Iyer, Lakshminarayan M

    2015-01-01

    Catalysis of NAD(+)-dependent ADP-ribosylation of proteins, nucleic acids, or small molecules has evolved in at least three structurally unrelated superfamilies of enzymes, namely ADP-ribosyltransferase (ART), the Sirtuins, and probably TM1506. Of these, the ART superfamily is the most diverse in terms of structure, active site residues, and targets that they modify. The primary diversification of the ART superfamily occurred in the context of diverse bacterial conflict systems, wherein ARTs play both offensive and defensive roles. These include toxin-antitoxin systems, virus-host interactions, intraspecific antagonism (polymorphic toxins), symbiont/parasite effectors/toxins, resistance to antibiotics, and repair of RNAs cleaved in conflicts. ARTs evolving in these systems have been repeatedly acquired by lateral transfer throughout eukaryotic evolution, starting from the PARP family, which was acquired prior to the last eukaryotic common ancestor. They were incorporated into eukaryotic regulatory/epigenetic control systems (e.g., PARP family and NEURL4), and also used as defensive (e.g., pierisin and CARP-1 families) or immunity-related proteins (e.g., Gig2-like ARTs). The ADP-ribosylation system also includes other domains, such as the Macro, ADP-ribosyl glycohydrolase, NADAR, and ADP-ribosyl cyclase, which appear to have initially diversified in bacterial conflict-related systems. Unlike ARTs, sirtuins appear to have a much smaller presence in conflict-related systems.

  4. The natural history of ADP-ribosyltransferases and the ADP-ribosylation system.

    Science.gov (United States)

    Aravind, L; Zhang, Dapeng; de Souza, Robson F; Anand, Swadha; Iyer, Lakshminarayan M

    2015-01-01

    Catalysis of NAD(+)-dependent ADP-ribosylation of proteins, nucleic acids, or small molecules has evolved in at least three structurally unrelated superfamilies of enzymes, namely ADP-ribosyltransferase (ART), the Sirtuins, and probably TM1506. Of these, the ART superfamily is the most diverse in terms of structure, active site residues, and targets that they modify. The primary diversification of the ART superfamily occurred in the context of diverse bacterial conflict systems, wherein ARTs play both offensive and defensive roles. These include toxin-antitoxin systems, virus-host interactions, intraspecific antagonism (polymorphic toxins), symbiont/parasite effectors/toxins, resistance to antibiotics, and repair of RNAs cleaved in conflicts. ARTs evolving in these systems have been repeatedly acquired by lateral transfer throughout eukaryotic evolution, starting from the PARP family, which was acquired prior to the last eukaryotic common ancestor. They were incorporated into eukaryotic regulatory/epigenetic control systems (e.g., PARP family and NEURL4), and also used as defensive (e.g., pierisin and CARP-1 families) or immunity-related proteins (e.g., Gig2-like ARTs). The ADP-ribosylation system also includes other domains, such as the Macro, ADP-ribosyl glycohydrolase, NADAR, and ADP-ribosyl cyclase, which appear to have initially diversified in bacterial conflict-related systems. Unlike ARTs, sirtuins appear to have a much smaller presence in conflict-related systems. PMID:25027823

  5. ADP-ribosylation of histones by ARTD1: an additional module of the histone code?

    Science.gov (United States)

    Hottiger, Michael O

    2011-06-01

    ADP-ribosylation is a covalent post-translational protein modification catalyzed by ADP-ribosyltransferases and is involved in important processes such as cell cycle regulation, DNA damage response, replication or transcription. Histones are ADP-ribosylated by ADP-ribosyltransferase diphtheria toxin-like 1 at specific amino acid residues, in particular lysines, of the histones tails. Specific ADP-ribosyl hydrolases and poly-ADP-ribose glucohydrolases degrade the ADP-ribose polymers. The ADP-ribose modification is read by zinc finger motifs or macrodomains, which then regulate chromatin structure and transcription. Thus, histone ADP-ribosylation may be considered an additional component of the histone code.

  6. Bacillus cereus Certhrax ADP-ribosylates vinculin to disrupt focal adhesion complexes and cell adhesion.

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    Simon, Nathan C; Barbieri, Joseph T

    2014-04-11

    Bacillus cereus is often associated with mild to moderate gastroenteritis; however, some recent isolates cause inhalational anthrax-like diseases and death. These potential emerging human pathogens express multiple virulence factors. B. cereus strain G9241 expresses anthrax toxin, several polysaccharide capsules, and the novel ADP-ribosyltransferase, Certhrax. In this study, we show that Certhrax ADP-ribosylates Arg-433 of vinculin, a protein that coordinates actin cytoskeleton and extracellular matrix interactions. ADP-ribosylation of vinculin disrupted focal adhesion complexes and redistributed vinculin to the cytoplasm. Exogenous vinculin rescued these phenotypes. This provides a mechanism for strain G9241 to breach host barrier defenses and promote bacterial growth and spread. Certhrax is the first bacterial toxin to add a post-translational modification to vinculin to disrupt the actin cytoskeleton.

  7. Structure of Plasmodium falciparum ADP-ribosylation factor 1

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    Cook, William J.; Smith, Craig D.; Senkovich, Olga; Holder, Anthony A.; Chattopadhyay, Debasish (UAB); (NIMR)

    2011-09-26

    Vesicular trafficking may play a crucial role in the pathogenesis and survival of the malaria parasite. ADP-ribosylation factors (ARFs) are among the major components of vesicular trafficking pathways in eukaryotes. The crystal structure of ARF1 GTPase from Plasmodium falciparum has been determined in the GDP-bound conformation at 2.5 {angstrom} resolution and is compared with the structures of mammalian ARF1s.

  8. Inhibiting poly(ADP-ribosylation) improves axon regeneration

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    Byrne, Alexandra B; McWhirter, Rebecca D; Sekine, Yuichi; Strittmatter, Stephen M; Miller, David M; Hammarlund, Marc

    2016-01-01

    The ability of a neuron to regenerate its axon after injury depends in part on its intrinsic regenerative potential. Here, we identify novel intrinsic regulators of axon regeneration: poly(ADP-ribose) glycohodrolases (PARGs) and poly(ADP-ribose) polymerases (PARPs). PARGs, which remove poly(ADP-ribose) from proteins, act in injured C. elegans GABA motor neurons to enhance axon regeneration. PARG expression is regulated by DLK signaling, and PARGs mediate DLK function in enhancing axon regeneration. Conversely, PARPs, which add poly(ADP-ribose) to proteins, inhibit axon regeneration of both C. elegans GABA neurons and mammalian cortical neurons. Furthermore, chemical PARP inhibitors improve axon regeneration when administered after injury. Our results indicate that regulation of poly(ADP-ribose) levels is a critical function of the DLK regeneration pathway, that poly-(ADP ribosylation) inhibits axon regeneration across species, and that chemical inhibition of PARPs can elicit axon regeneration. DOI: http://dx.doi.org/10.7554/eLife.12734.001

  9. The NarE protein of Neisseria gonorrhoeae catalyzes ADP-ribosylation of several ADP-ribose acceptors despite an N-terminal deletion.

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    Rodas, Paula I; Álamos-Musre, A Said; Álvarez, Francisca P; Escobar, Alejandro; Tapia, Cecilia V; Osorio, Eduardo; Otero, Carolina; Calderón, Iván L; Fuentes, Juan A; Gil, Fernando; Paredes-Sabja, Daniel; Christodoulides, Myron

    2016-09-01

    The ADP-ribosylating enzymes are encoded in many pathogenic bacteria in order to affect essential functions of the host. In this study, we show that Neisseria gonorrhoeae possess a locus that corresponds to the ADP-ribosyltransferase NarE, a previously characterized enzyme in N. meningitidis The 291 bp coding sequence of gonococcal narE shares 100% identity with part of the coding sequence of the meningococcal narE gene due to a frameshift previously described, thus leading to a 49-amino-acid deletion at the N-terminus of gonococcal NarE protein. However, we found a promoter region and a GTG start codon, which allowed expression of the protein as demonstrated by RT-PCR and western blot analyses. Using a gonococcal NarE-6xHis fusion protein, we demonstrated that the gonococcal enzyme underwent auto-ADP-ribosylation but to a lower extent than meningococcal NarE. We also observed that gonoccocal NarE exhibited ADP-ribosyltransferase activity using agmatine and cell-free host proteins as ADP-ribose acceptors, but its activity was inhibited by human β-defensins. Taken together, our results showed that NarE of Neisseria gonorrhoeae is a functional enzyme that possesses key features of bacterial ADP-ribosylating enzymes. PMID:27465490

  10. Arginine ADP-ribosylation mechanism based on structural snapshots of iota-toxin and actin complex

    OpenAIRE

    Tsurumura, Toshiharu; Tsumori, Yayoi; Qiu, Hao; Oda, Masataka; Sakurai, Jun; Nagahama, Masahiro; Tsuge, Hideaki

    2012-01-01

    Clostridium perfringens iota-toxin (Ia) mono-ADP ribosylates Arg177 of actin, leading to cytoskeletal disorganization and cell death. To fully understand the reaction mechanism of arginine-specific mono-ADP ribosyl transferase, the structure of the toxin-substrate protein complex must be characterized. Recently, we solved the crystal structure of Ia in complex with actin and the nonhydrolyzable NAD+ analog βTAD (thiazole-4-carboxamide adenine dinucleotide); however, the structures of the NAD+...

  11. ADP-ribosylation of dinitrogenase reductase in Azospirillum brasilense is regulated by AmtB-dependent membrane sequestration of DraG.

    Science.gov (United States)

    Huergo, Luciano F; Souza, Emanuel M; Araujo, Mariana S; Pedrosa, Fábio O; Chubatsu, Leda S; Steffens, Maria B R; Merrick, Mike

    2006-01-01

    Nitrogen fixation in some diazotrophic bacteria is regulated by mono-ADP-ribosylation of dinitrogenase reductase (NifH) that occurs in response to addition of ammonium to the extracellular medium. This process is mediated by dinitrogenase reductase ADP-ribosyltransferase (DraT) and reversed by dinitrogenase reductase glycohydrolase (DraG), but the means by which the activities of these enzymes are regulated are unknown. We have investigated the role of the P(II) proteins (GlnB and GlnZ), the ammonia channel protein AmtB and the cellular localization of DraG in the regulation of the NifH-modification process in Azospirillum brasilense. GlnB, GlnZ and DraG were all membrane-associated after an ammonium shock, and both this membrane sequestration and ADP-ribosylation of NifH were defective in an amtB mutant. We now propose a model in which membrane association of DraG after an ammonium shock creates a physical separation from its cytoplasmic substrate NifH thereby inhibiting ADP-ribosyl-removal. Our observations identify a novel role for an ammonia channel (Amt) protein in the regulation of bacterial nitrogen metabolism by mediating membrane sequestration of a protein other than a P(II) family member. They also suggest a model for control of ADP-ribosylation that is likely to be applicable to all diazotrophs that exhibit such post-translational regulation of nitrogenase.

  12. Detection and Quantification of ADP-Ribosylated RhoA/B by Monoclonal Antibody

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    Rohrbeck, Astrid; Fühner, Viola; Schröder, Anke; Hagemann, Sandra; Vu, Xuan-Khang; Berndt, Sarah; Hust, Michael; Pich, Andreas; Just, Ingo

    2016-01-01

    Clostridium botulinum exoenzyme C3 is the prototype of C3-like ADP-ribosyltransferases that modify the GTPases RhoA, B, and C. C3 catalyzes the transfer of an ADP-ribose moiety from the co-substrate nicotinamide adenine dinucleotide (NAD) to asparagine-41 of Rho-GTPases. Although C3 does not possess cell-binding/-translocation domains, C3 is able to efficiently enter intact cells, including neuronal and macrophage-like cells. Conventionally, the detection of C3 uptake into cells is carried out via the gel-shift assay of modified RhoA. Since this gel-shift assay does not always provide clear, evaluable results an additional method to confirm the ADP-ribosylation of RhoA is necessary. Therefore, a new monoclonal antibody has been generated that specifically detects ADP-ribosylated RhoA/B, but not RhoC, in Western blot and immunohistochemical assay. The scFv antibody fragment was selected by phage display using the human naive antibody gene libraries HAL9/10. Subsequently, the antibody was produced as scFv-Fc and was found to be as sensitive as a commercially available RhoA antibody providing reproducible and specific results. We demonstrate that this specific antibody can be successfully applied for the analysis of ADP-ribosylated RhoA/B in C3-treated Chinese hamster ovary (CHO) and HT22 cells. Moreover, ADP-ribosylation of RhoA was detected within 10 min in C3-treated CHO wild-type cells, indicative of C3 cell entry. PMID:27043630

  13. Evidence that phospholipase D mediates ADP ribosylation factor- dependent formation of Golgi coated vesicles

    OpenAIRE

    1996-01-01

    Formation of coatomer-coated vesicles from Golgi-enriched membranes requires the activation of a small GTP-binding protein, ADP ribosylation factor (ARF). ARF is also an efficacious activator of phospholipase D (PLD), an activity that is relatively abundant on Golgi- enriched membranes. It has been proposed that ARF, which is recruited onto membranes from cytosolic pools, acts directly to promote coatomer binding and is in a 3:1 stoichiometry with coatomer on coated vesicles. We present evide...

  14. Poly(ADP-ribosyl)ation is recognized by ECT2 during mitosis.

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    Li, Mo; Bian, Chunjing; Yu, Xiaochun

    2014-01-01

    Poly(ADP-ribosyl)ation is an unique posttranslational modification and required for spindle assembly and function during mitosis. However, the molecular mechanism of poly(ADP-ribose) (PAR) in mitosis remains elusive. Here, we show the evidence that PAR is recognized by ECT2, a key guanine nucleotide exchange factor in mitosis. The BRCT domain of ECT2 directly binds to PAR both in vitro and in vivo. We further found that α-tubulin is PARylated during mitosis. PARylation of α-tubulin is recognized by ECT2 and recruits ECT2 to mitotic spindle for completing mitosis. Taken together, our study reveals a novel mechanism by which PAR regulates mitosis.

  15. Poly(ADP-ribosyl)ation is recognized by ECT2 during mitosis

    OpenAIRE

    Li, Mo; Bian, Chunjing; Yu, Xiaochun

    2014-01-01

    Poly(ADP-ribosyl)ation is an unique posttranslational modification and required for spindle assembly and function during mitosis. However, the molecular mechanism of poly(ADP-ribose) (PAR) in mitosis remains elusive. Here, we show the evidence that PAR is recognized by ECT2, a key guanine nucleotide exchange factor in mitosis. The BRCT domain of ECT2 directly binds to PAR both in vitro and in vivo. We further found that α-tubulin is PARylated during mitosis. PARylation of α-tubulin is recogni...

  16. Class I ADP-ribosylation factors are involved in enterovirus 71 replication.

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    Jianmin Wang

    Full Text Available Enterovirus 71 is one of the major causative agents of hand, foot, and mouth disease in infants and children. Replication of enterovirus 71 depends on host cellular factors. The viral replication complex is formed in novel, cytoplasmic, vesicular compartments. It has not been elucidated which cellular pathways are hijacked by the virus to create these vesicles. Here, we investigated whether proteins associated with the cellular secretory pathway were involved in enterovirus 71 replication. We used a loss-of-function assay, based on small interfering RNA. We showed that enterovirus 71 RNA replication was dependent on the activity of Class I ADP-ribosylation factors. Simultaneous depletion of ADP-ribosylation factors 1 and 3, but not three others, inhibited viral replication in cells. We also demonstrated with various techniques that the brefeldin-A-sensitive guanidine nucleotide exchange factor, GBF1, was critically important for enterovirus 71 replication. Our results suggested that enterovirus 71 replication depended on GBF1-mediated activation of Class I ADP-ribosylation factors. These results revealed a connection between enterovirus 71 replication and the cellular secretory pathway; this pathway may represent a novel target for antiviral therapies.

  17. Brefeldin A-induced ADP-ribosylation in the structure and function of the Golgi complex.

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    Colanzi, A; Mironov, A; Weigert, R; Limina, C; Flati, S; Cericola, C; Di Tullio, G; Di Girolamo, M; Corda, D; De Matteis, M A; Luini, A

    1997-01-01

    Brefeldin A (BFA) is a fungal metabolite that exerts generally inhibitory actions on membrane transport and induces the disappearance of the Golgi complex. Previously we have shown that BFA stimulates the ADP-ribosylation of two cytosolic proteins of 38 and 50 KD. The BFA-binding components mediating the BFA-sensitive ADP-ribosylation (BAR) and the effect of BFA on ARF binding to Golgi membranes have similar specificities and affinities for BFA and its analogues, suggesting that BAR may have a role in the cellular effects of BFA. To investigate this we used the approach to impair BAR activity by the use of BAR inhibitors. A series of BAR inhibitors was developed and their effects were studied in RBL cells treated with BFA. In addition to the common ADP-ribosylation inhibitors (nicotinamide and aminobenzamide), compounds belonging to the cumarin (novobiocin, cumermycin, dicumarol) class were active BAR inhibitors. All BAR inhibitors were able to prevent the BFA-induced redistribution of a Golgi marker (Helix pomatia lectin) into the endoplasmic reticulum, as assessed in immunofluorescence experiments. At the ultrastructural level, BAR inhibitors prevented the tubular-vesicular transformation of the Golgi complex caused by BFA. The potencies of these compounds in preventing the BFA effects on the Golgi complex were similar to those at which they inhibited BAR. Altogether these data support the hypothesis that BAR mediates at least some of the effects of BFA on the Golgi structure and function. PMID:9193673

  18. Regulation of chromatin structure by poly(ADP-ribosylation

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    Sascha eBeneke

    2012-09-01

    Full Text Available The interaction of DNA with proteins in the context of chromatin has to be tightly regulated to achieve so different tasks as packaging, transcription, replication and repair. The very rapid and transient post-translational modification of proteins by poly(ADP-ribose has been shown to take part in all four. Originally identified as immediate cellular answer to a variety of genotoxic stresses, already early data indicated the ability of this highly charged nucleic acid-like polymer to modulate nucleosome structure, the basic unit of chromatin. At the same time the enzyme responsible for synthesizing poly(ADP-ribose, the zinc-finger protein poly(ADP-ribose polymerase-1 (PARP1, was shown to control transcription initiation as basic factor TFIIC within the RNA-polymerase II machinery. Later research focused more on PARP-mediated regulation of DNA repair and cell death, but in the last few years, transcription as well as chromatin modulation has re-appeared on the scene. This review will discuss the impact of PARP1 on transcription and transcription factors, its implication in chromatin remodeling for DNA repair and probably also replication, and its role in controlling epigenetic events such as DNA methylation and the functionality of the insulator protein CCCTC-binding factor.

  19. Poly(ADP-ribosyl)ation-dependent Transient Chromatin Decondensation and Histone Displacement following Laser Microirradiation.

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    Strickfaden, Hilmar; McDonald, Darin; Kruhlak, Michael J; Haince, Jean-Francois; Th'ng, John P H; Rouleau, Michele; Ishibashi, Toytaka; Corry, Gareth N; Ausio, Juan; Underhill, D Alan; Poirier, Guy G; Hendzel, Michael J

    2016-01-22

    Chromatin undergoes a rapid ATP-dependent, ATM and H2AX-independent decondensation when DNA damage is introduced by laser microirradiation. Although the detailed mechanism of this decondensation remains to be determined, the kinetics of decondensation are similar to the kinetics of poly(ADP-ribosyl)ation. We used laser microirradiation to introduce DNA strand breaks into living cells expressing a photoactivatable GFP-tagged histone H2B. We find that poly(ADP-ribosyl)ation mediated primarily by poly(ADP-ribose) polymerase 1 (PARP1) is responsible for the rapid decondensation of chromatin at sites of DNA damage. This decondensation of chromatin correlates temporally with the displacement of histones, which is sensitive to PARP inhibition and is transient in nature. Contrary to the predictions of the histone shuttle hypothesis, we did not find that histone H1 accumulated on poly(ADP-ribose) (PAR) in vivo. Rather, histone H1, and to a lessor extent, histones H2A and H2B were rapidly depleted from the sites of PAR accumulation. However, histone H1 returns to chromatin and the chromatin recondenses. Thus, the PARP-dependent relaxation of chromatin closely correlates with histone displacement.

  20. Roles of Asp179 and Glu270 in ADP-Ribosylation of Actin by Clostridium perfringens Iota Toxin.

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    Alexander Belyy

    Full Text Available Clostridium perfringens iota toxin is a binary toxin composed of the enzymatically active component Ia and receptor binding component Ib. Ia is an ADP-ribosyltransferase, which modifies Arg177 of actin. The previously determined crystal structure of the actin-Ia complex suggested involvement of Asp179 of actin in the ADP-ribosylation reaction. To gain more insights into the structural requirements of actin to serve as a substrate for toxin-catalyzed ADP-ribosylation, we engineered Saccharomyces cerevisiae strains, in which wild type actin was replaced by actin variants with substitutions in residues located on the Ia-actin interface. Expression of the actin mutant Arg177Lys resulted in complete resistance towards Ia. Actin mutation of Asp179 did not change Ia-induced ADP-ribosylation and growth inhibition of S. cerevisiae. By contrast, substitution of Glu270 of actin inhibited the toxic action of Ia and the ADP-ribosylation of actin. In vitro transcribed/translated human β-actin confirmed the crucial role of Glu270 in ADP-ribosylation of actin by Ia.

  1. Rapamycin inhibits poly(ADP-ribosyl)ation in intact cells

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    Fahrer, Joerg, E-mail: joerg.fahrer@uni-ulm.de [Molecular Toxicology Group, Department of Biology, University of Konstanz (Germany); Wagner, Silvia [Clinic of General, Visceral- and Transplantation Surgery, ZMF, University Hospital Tuebingen (Germany); Buerkle, Alexander [Molecular Toxicology Group, Department of Biology, University of Konstanz (Germany); Koenigsrainer, Alfred [Clinic of General, Visceral- and Transplantation Surgery, ZMF, University Hospital Tuebingen (Germany)

    2009-08-14

    Rapamycin is an immunosuppressive drug, which inhibits the mammalian target of rapamycin (mTOR) kinase activity inducing changes in cell proliferation. Synthesis of poly(ADP-ribose) (PAR) is an immediate cellular response to genotoxic stress catalyzed mostly by poly(ADP-ribose) polymerase 1 (PARP-1), which is also controlled by signaling pathways. Therefore, we investigated whether rapamycin affects PAR production. Strikingly, rapamycin inhibited PAR synthesis in living fibroblasts in a dose-dependent manner as monitored by immunofluorescence. PARP-1 activity was then assayed in vitro, revealing that down-regulation of cellular PAR production by rapamycin was apparently not due to competitive PARP-1 inhibition. Further studies showed that rapamycin did not influence the cellular NAD pool and the activation of PARP-1 in extracts of pretreated fibroblasts. Collectively, our data suggest that inhibition of cellular PAR synthesis by rapamycin is mediated by formation of a detergent-sensitive complex in living cells, and that rapamycin may have a potential as therapeutic PARP inhibitor.

  2. Endogenous ADP-ribosylation of elongation factor 2 in polyoma virus-transformed baby hamster kidney cells

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    Fendrick, J.L.; Iglewski, W.J. (Univ. of Rochester, NY (USA))

    1989-01-01

    Polyoma virus-transformed baby hamster kidney (pyBHK) cells were cultured in medium containing ({sup 32}P)orthophosphate and 105 (vol/vol) fetal bovine serum. A {sup 32}P-labeled protein with an apparent molecular mass of 97 kDa was immunoprecipitated from cell lysates with antiserum to ADP-ribosylated elongation factor 2 (EF-2). The {sup 32}P labeling of the protein was enhanced by culturing cells in medium containing 2% serum instead of 10% serum. The {sup 32}P label was completely removed from the protein by treatment with snake venom phosphodiesterase and the digestion product was identified as ({sup 32}P)AMP, indicating the protein was mono-ADP-ribosylated. HPLC analysis of tryptic peptides of the {sup 32}P-labeled 97-kDa protein and purified EF-2, which was ADP-ribosylated in vitro with diphtheria toxin fragment A and ({sup 32}P)NAD, demonstrated an identical labeled peptide in the two proteins. The data strongly suggest that EF-2 was endogenously ADP-ribosylated in pyBHK cells. Maximum incorporation of radioactivity in EF-2 occurred by 12 hr and remained constant over the subsequent 12 hr. It was estimated that 30-35% of the EF-2 was ADP-ribosylated in cells cultured in medium containing 2% serum. When {sup 32}P-labeled cultures were incubated in medium containing unlabeled phosphate, the {sup 32}P label was lost from the EF-2 within 30 min.

  3. Guanine nucleotide-binding proteins that enhance choleragen ADP-ribosyltransferase activity: nucleotide and deduced amino acid sequence of an ADP-ribosylation factor cDNA.

    OpenAIRE

    Price, S R; Nightingale, M.; Tsai, S C; Williamson, K. C.; Adamik, R; H. C. Chen; Moss, J; M. Vaughan

    1988-01-01

    Three (two soluble and one membrane) guanine nucleotide-binding proteins (G proteins) that enhance ADP-ribosylation of the Gs alpha stimulatory subunit of the adenylyl cyclase (EC 4.6.1.1) complex by choleragen have recently been purified from bovine brain. To further define the structure and function of these ADP-ribosylation factors (ARFs), we isolated a cDNA clone (lambda ARF2B) from a bovine retinal library by screening with a mixed heptadecanucleotide probe whose sequence was based on th...

  4. NMR resonance assignments of NarE, a putative ADP-ribosylating toxin from Neisseria meningitidis

    NARCIS (Netherlands)

    Carlier, L.P.A.; Köhler, Christian; Veggi, D.; Pizza, M.; Soriani, M.; Boelens, R.; Bonvin, A.M.J.J.

    2011-01-01

    NarE is a 16 kDa protein identified from Neisseria meningitidis, one of the bacterial pathogens responsible for meningitis. NarE belongs to the ADP-ribosyltransferase family and catalyses the transfer of ADP-ribose moieties to arginine residues in target protein acceptors. Many pathogenic bacteria u

  5. Poly(ADP-ribosylation) regulates chromatin organization through histone H3 modification and DNA methylation of the first cell cycle of mouse embryos

    Energy Technology Data Exchange (ETDEWEB)

    Osada, Tomoharu, E-mail: osada.tomoharu@mg.medience.co.jp [Advanced Medical Science Research Department, Mitsubishi Chemical Medience Corporation, 14-1 Sunayama, Kamisu-shi, Ibaragi 314-0255 (Japan); Department of Regenerative and Developmental Biology, Mitsubishi Kagaku Institute of Life Sciences (MITILS), 11 Minamiooya, Machida-shi, Tokyo 194-8511 (Japan); Rydén, Anna-Margareta [Division of Genome Stability Research, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045 (Japan); Masutani, Mitsuko, E-mail: mmasutan@ncc.go.jp [Division of Genome Stability Research, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045 (Japan)

    2013-04-26

    Highlights: •Histone modification of the mouse pronuclei is regulated by poly(ADP-ribosylation). •Hypermethylation of the mouse female pronuclei is maintained by poly(ADP-ribosylation). •Parp1 is physically interacted with Suz12, which may function in the pronuclei. •Poly(ADP-ribosylation) affects ultrastructure of chromatin of the mouse pronucleus. -- Abstract: We examined the roles of poly(ADP-ribosylation) in chromatin remodeling during the first cell cycle of mouse embryos. Drug-based inhibition of poly(ADP-ribosylation) by a PARP inhibitor, PJ-34, revealed up-regulation of dimethylation of histone H3 at lysine 4 in male pronuclei and down-regulation of dimethylation of histone H3 at lysine 9 (H3K9) and lysine 27 (H3K27). Association of poly(ADP-ribosylation) with histone modification was suggested to be supported by the interaction of Suz12, a histone methyltransferase in the polycomb complex, with Parp1. PARP activity was suggested to be required for a proper localization and maintenance of Suz12 on chromosomes. Notably, DNA methylation level of female pronuclei in one-cell embryos was robustly decreased by PJ-34. Electron microscopic analysis showed a frequent appearance of unusual electron-dense areas within the female pronuclei, implying the disorganized and hypercondensed chromatin ultrastructure. These results show that poly(ADP-ribosylation) is important for the integrity of non-equivalent epigenetic dynamics of pronuclei during the first cell cycle of mouse embryos.

  6. Poly-ADP-ribosylation of HMGB1 regulates TNFSF10/TRAIL resistance through autophagy.

    Science.gov (United States)

    Yang, Minghua; Liu, Liying; Xie, Min; Sun, Xiaofang; Yu, Yan; Kang, Rui; Yang, Liangchun; Zhu, Shan; Cao, Lizhi; Tang, Daolin

    2015-01-01

    Both apoptosis ("self-killing") and autophagy ("self-eating") are evolutionarily conserved processes, and their crosstalk influences anticancer drug sensitivity and cell death. However, the underlying mechanism remains unclear. Here, we demonstrated that HMGB1 (high mobility group box 1), normally a nuclear protein, is a crucial regulator of TNFSF10/TRAIL (tumor necrosis factor [ligand] superfamily, member 10)-induced cancer cell death. Activation of PARP1 (poly [ADP-ribose] polymerase 1) was required for TNFSF10-induced ADP-ribosylation of HMGB1 in cancer cells. Moreover, pharmacological inhibition of PARP1 activity or knockdown of PARP1 gene expression significantly inhibited TNFSF10-induced HMGB1 cytoplasmic translocation and subsequent HMGB1-BECN1 complex formation. Furthermore, suppression of the PARP1-HMGB1 pathway diminished autophagy, increased apoptosis, and enhanced the anticancer activity of TNFSF10 in vitro and in a subcutaneous tumor model. These results indicate that PARP1 acts as a prominent upstream regulator of HMGB1-mediated autophagy and maintains a homeostatic balance between apoptosis and autophagy, which provides new insight into the mechanism of TNFSF10 resistance.

  7. Mice lacking the ADP ribosyl cyclase CD38 exhibit attenuated renal vasoconstriction to angiotensin II, endothelin-1, and norepinephrine

    OpenAIRE

    Thai, Tiffany L.; Arendshorst, William J.

    2009-01-01

    ADP ribosyl (ADPR) cyclases comprise a family of ectoenzymes recently shown to influence cytosolic Ca2+ concentration in a variety of cell types. At least two ADPR cyclase family members have been identified in mammals: CD38 and CD157. We recently found reduced renal vascular reactivity to angiotensin II (ANG II), endothelin-1 (ET-1), and norepinephrine (NE) in the presence of the broad ADPR cyclase inhibitor nicotinamide. We hypothesized that CD38 mediates effects attributed to ADPR cyclase....

  8. Modes of action of ADP-ribosylated elongation factor 2 in inhibiting the polypeptide elongation cycle: a modeling study.

    Directory of Open Access Journals (Sweden)

    Kevin C Chen

    Full Text Available Despite the fact that ADP-ribosylation of eukaryotic elongation factor 2 (EF2 leads to inhibition of protein synthesis, the mechanism by which ADP-ribosylated EF2 (ADPR•EF2 causes this inhibition remains controversial. Here, we applied modeling approaches to investigate the consequences of various modes of ADPR•EF2 inhibitory actions on the two coupled processes, the polypeptide chain elongation and ADP-ribosylation of EF2. Modeling of experimental data indicates that ADPR•EF2 fully blocks the late-phase translocation of tRNAs; but the impairment in the translocation upstream process, mainly the GTP-dependent factor binding with the pretranslocation ribosome and/or the guanine nucleotide exchange in EF2, is responsible for the overall inhibition kinetics. The reduced ADPR•EF2-ribosome association spares the ribosome to bind and shield native EF2 against toxin attack, thereby deferring the inhibition of protein synthesis inhibition and inactivation of EF2. Minimum association with the ribosome also keeps ADPR•EF2 in an accessible state for toxins to catalyze the reverse reaction when nicotinamide becomes available. Our work underscores the importance of unveiling the interactions between ADPR•EF2 and the ribosome, and argues against that toxins inhibit protein synthesis through converting native EF2 to a competitive inhibitor to actively disable the ribosome.

  9. mADP-RTs: Versatile virulence factors from bacterial pathogens of plants and mammals

    Directory of Open Access Journals (Sweden)

    Lennart eWirthmueller

    2012-06-01

    Full Text Available Mono ADP-ribosyltransferases (mADP-RTs are a family of enzymes that cleave NAD+ and covalently attach the ADP-ribosyl moiety to target proteins. mADP-RTs are well established as important virulence factors of bacteria that infect mammals. Cholera toxin, pertussis toxin and diphteria toxin are three of the best-known examples of mADP-RTs. They modify host target proteins in order to promote infection and/or killing of the host cell. Despite low sequence similarity at the primary amino acid level, mADP-RTs share a conserved core catalytic fold and structural biology has made important contributions to elucidating how mADP-RTs modify mammalian host targets. Recently, mADP-RTs were shown to be present in plant pathogenic bacteria, suggesting that mADP-RTs are also important virulence factors of plant pathogens. Crystal structures of plant pathogenic bacterial mADP-RTs are also now available. Here we review the structure/function of mADP-RTs from pathogens of mammals and plants, highlighting both commonalities and differences.

  10. ADP ribosylation factor like 2 (Arl2 regulates breast tumor aggressivity in immunodeficient mice.

    Directory of Open Access Journals (Sweden)

    Anne Beghin

    Full Text Available We have previously reported that ADP ribosylation factor like 2 (Arl2, a small GTPase, content influences microtubule dynamics and cell cycle distribution in breast tumor cells, as well as the degree and distribution of phosphorylated P53. Here we show, in two different human breast adenocarcinoma models, that Arl2 content has a major impact on breast tumor cell aggressivity both in vitro and in vivo. Cells with reduced content of Arl2 displayed reduced contact inhibition, increased clonogenic or cluster formation as well as a proliferative advantage over control cells in an in vitro competition assay. These cells also caused larger tumors in SCID mice, a phenotype which was mimicked by the in vivo administration of siRNA directed against Arl2. Cells with increased Arl2 content displayed reduced aggressivity, both in vitro and in vivo, with enhanced necrosis and were also found to contain increased PP2A phosphatase activity. A rt-PCR analysis of fresh human tumor breast samples suggested that low Arl2 expression was associated with larger tumor size and greater risk of lymph node involvement at diagnosis. These data underline the role of Arl2, a small GTPase, as an important regulator of breast tumor cell aggressivity, both in vitro and in vivo.

  11. MARTX effector cross kingdom activation by Golgi-associated ADP-ribosylation factors.

    Science.gov (United States)

    Kim, Byoung Sik; Satchell, Karla J F

    2016-08-01

    Vibrio vulnificus infects humans and causes lethal septicemia. The primary virulence factor is a multifunctional-autoprocessing repeats-in-toxin (MARTX) toxin consisting of conserved repeats-containing regions and various effector domains. Recent genomic analyses for the newly emerged V. vulnificus biotype 3 strain revealed that its MARTX toxin has two previously unknown effector domains. Herein, we characterized one of these domains, Domain X (DmXVv ). A structure-based homology search revealed that DmXVv belongs to the C58B cysteine peptidase subfamily. When ectopically expressed in cells, DmXVv was autoprocessed and induced cytopathicity including Golgi dispersion. When the catalytic cysteine or the region flanking the scissile bond was mutated, both autoprocessing and cytopathicity were significantly reduced indicating that DmXVv cytopathicity is activated by amino-terminal autoprocessing. Consistent with this, host cell protein export was affected by Vibrio cells producing a toxin with wild-type, but not catalytically inactive, DmXVv . DmXVv was found to localize to Golgi and to directly interact with Golgi-associated ADP-ribosylation factors ARF1, ARF3 and ARF4, although ARF binding was not necessary for the subcellular localization. Rather, this interaction was found to induce autoprocessing of DmXVv . These data demonstrate that the V. vulnificus hijacks the host ARF proteins to activate the cytopathic DmXVv effector domain of MARTX toxin. PMID:26780191

  12. Cholix Toxin, a Novel ADP-ribosylating Factor from Vibrio cholerae

    Energy Technology Data Exchange (ETDEWEB)

    Jorgensen, Rene; Purdy, Alexandra E.; Fieldhouse, Robert J.; Kimber, Matthew S.; Bartlett, Douglas H.; Merrill, A. Rod (Guelph); (NIH); (UCSD)

    2008-07-15

    The ADP-ribosyltransferases are a class of enzymes that display activity in a variety of bacterial pathogens responsible for causing diseases in plants and animals, including those affecting mankind, such as diphtheria, cholera, and whooping cough. We report the characterization of a novel toxin from Vibrio cholerae, which we call cholix toxin. The toxin is active against mammalian cells (IC50 = 4.6 {+-} 0.4 ng/ml) and crustaceans (Artemia nauplii LD50 = 10 {+-} 2 {mu}g/ml). Here we show that this toxin is the third member of the diphthamide-specific class of ADP-ribose transferases and that it possesses specific ADP-ribose transferase activity against ribosomal eukaryotic elongation factor 2. We also describe the high resolution crystal structures of the multidomain toxin and its catalytic domain at 2.1- and 1.25-{angstrom} resolution, respectively. The new structural data show that cholix toxin possesses the necessary molecular features required for infection of eukaryotes by receptor-mediated endocytosis, translocation to the host cytoplasm, and inhibition of protein synthesis by specific modification of elongation factor 2. The crystal structures also provide important insight into the structural basis for activation of toxin ADP-ribosyltransferase activity. These results indicate that cholix toxin may be an important virulence factor of Vibrio cholerae that likely plays a significant role in the survival of the organism in an aquatic environment.

  13. ADP-ribosylation of actins in fibroblasts and myofibroblasts by botulinum C2 toxin: Influence on microfilament morphology and migratory behavior

    DEFF Research Database (Denmark)

    Rønnov-Jessen, Lone; Petersen, Ole William

    1996-01-01

    botulinum C2 toxin. The substrate for C2 toxin is globular actin, which upon ribosylation cannot incorporate into microfilaments. The pattern of actin ADP-ribosylation in (myo)fibroblasts in the presence of [32P]NAD was analyzed by isoelectric focusing, fluorography and immunoblotting. The influence of C2...... toxin on microfilaments in intact cells was further assessed by immunofluorescence, and motility was measured in a mass migration assay and by computerized video time-lapse microscopy. We show here that C2 toxin specifically ribosylates - and -actin in both fibroblasts and myofibroblasts. Whereas...

  14. Structure of an ADP-ribosylation factor, ARF1, from Entamoeba histolytica bound to Mg(2+)-GDP.

    Science.gov (United States)

    Serbzhinskiy, Dmitry A; Clifton, Matthew C; Sankaran, Banumathi; Staker, Bart L; Edwards, Thomas E; Myler, Peter J

    2015-05-01

    Entamoeba histolytica is the etiological agent of amebiasis, a diarrheal disease which causes amoebic liver abscesses and amoebic colitis. Approximately 50 million people are infected worldwide with E. histolytica. With only 10% of infected people developing symptomatic amebiasis, there are still an estimated 100,000 deaths each year. Because of the emergence of resistant strains of the parasite, it is necessary to find a treatment which would be a proper response to this challenge. ADP-ribosylation factor (ARF) is a member of the ARF family of GTP-binding proteins. These proteins are ubiquitous in eukaryotic cells; they generally associate with cell membranes and regulate vesicular traffic and intracellular signalling. The crystal structure of ARF1 from E. histolytica has been determined bound to magnesium and GDP at 1.8 Å resolution. Comparison with other structures of eukaryotic ARF proteins shows a highly conserved structure and supports the interswitch toggle mechanism of communicating the conformational state to partner proteins.

  15. PARP-2 regulates cell cycle-related genes through histone deacetylation and methylation independently of poly(ADP-ribosyl)ation

    Energy Technology Data Exchange (ETDEWEB)

    Liang, Ya-Chen; Hsu, Chiao-Yu [Institute of Molecular Biology, National Chung Hsing University, Taichung 40227, Taiwan (China); Yao, Ya-Li [Department of Biotechnology, Asia University, Taichung 41354, Taiwan (China); Yang, Wen-Ming, E-mail: yangwm@nchu.edu.tw [Institute of Molecular Biology, National Chung Hsing University, Taichung 40227, Taiwan (China)

    2013-02-01

    Highlights: ► PARP-2 acts as a transcription co-repressor independently of PARylation activity. ► PARP-2 recruits HDAC5, 7, and G9a and generates repressive chromatin. ► PARP-2 is recruited to the c-MYC promoter by DNA-binding factor YY1. ► PARP-2 represses cell cycle-related genes and alters cell cycle progression. -- Abstract: Poly(ADP-ribose) polymerase-2 (PARP-2) catalyzes poly(ADP-ribosyl)ation (PARylation) and regulates numerous nuclear processes, including transcription. Depletion of PARP-2 alters the activity of transcription factors and global gene expression. However, the molecular action of how PARP-2 controls the transcription of target promoters remains unclear. Here we report that PARP-2 possesses transcriptional repression activity independently of its enzymatic activity. PARP-2 interacts and recruits histone deacetylases HDAC5 and HDAC7, and histone methyltransferase G9a to the promoters of cell cycle-related genes, generating repressive chromatin signatures. Our findings propose a novel mechanism of PARP-2 in transcriptional regulation involving specific protein–protein interactions and highlight the importance of PARP-2 in the regulation of cell cycle progression.

  16. An ADP-Ribosylation Factor GTPase-activating Protein Git2-short/KIAA0148 Is Involved in Subcellular Localization of Paxillin and Actin Cytoskeletal Organization

    OpenAIRE

    Mazaki, Yuichi; Hashimoto, Shigeru; Okawa, Katsuya; Tsubouchi, Asako; Nakamura, Kuniaki; Yagi, Ryohei; Yano, Hajime; Kondo, Akiko; Iwamatsu, Akihiro; Mizoguchi, Akira; Sabe, Hisataka

    2001-01-01

    Paxillin acts as an adaptor protein in integrin signaling. We have shown that paxillin exists in a relatively large cytoplasmic pool, including perinuclear areas, in addition to focal complexes formed at the cell periphery and focal adhesions formed underneath the cell. Several ADP-ribosylation factor (ARF) GTPase-activating proteins (GAPs; ARFGAPs) have been shown to associate with paxillin. We report here that Git2-short/KIAA0148 exhibits properties of a paxillin...

  17. BIG2, A Guanine Nucleotide Exchange Factor for ADP-Ribosylation Factors: Its Localization to Recycling Endosomes and Implication in the Endosome IntegrityD⃞

    OpenAIRE

    Shin, Hye-Won; Morinaga, Naoko; NODA, Masatoshi; Nakayama, Kazuhisa

    2004-01-01

    Small GTPases of the ADP-ribosylation factor (ARF) family play a key role in membrane trafficking by regulating coated vesicle formation, and guanine nucleotide exchange is essential for the ARF function. Brefeldin A blocks the ARF-triggered coat assembly by inhibiting the guanine nucleotide exchange on ARFs and causes disintegration of the Golgi complex and tubulation of endosomal membranes. BIG2 is one of brefeldin A-inhibited guanine nucleotide exchange factors for the ARF GTPases and is a...

  18. ADP-ribosylation Factor-related Protein 1 Interacts with NS5A and Regulates Hepatitis C Virus Propagation

    Science.gov (United States)

    Lim, Yun-Sook; Ngo, Huong T. T.; Lee, Jihye; Son, Kidong; Park, Eun-Mee; Hwang, Soon B.

    2016-01-01

    The life cycle of hepatitis C virus (HCV) is tightly coupled to the lipid metabolism of host cells. In order to identify host factors involved in HCV propagation, we have previously screened a small interfering RNA (siRNA) library targeting host genes that control lipid metabolism and lipid droplet (LD) formation using cell culture-grown HCV (HCVcc)-infected cells. In this study, we selected and characterized the gene encoding ADP-ribosylation factor-related protein 1 (ARFRP1). ARFRP1 is essential for LD growth and is involved in the regulation of lipolysis. siRNA-mediated knockdown of ARFRP1 significantly inhibited HCV replication in both subgenomic replicon cells and HCVcc-infected cells. ARFRP1 interacted with NS5A and NS5A partially colocalized with LD. Silencing of ARFRP1 abrogated HCV-induced LD growth and viral protein expressions. Moreover, ARFRP1 recruited synaptosomal-associated protein 23 (SNAP23) to sites in close proximity to LDs in HCV-infected cells. Silencing of ARFRP1 ablated relocalization of SNAP23 to LD. These data indicate that HCV regulates ARFRP1 for LD growth to facilitate viral propagation and thus ARFRP1 may be a potential target for antiviral therapy. PMID:27550144

  19. Effect of meta-iodo-benzylguanine, inhibitor of mono (ADP-ribosylation), on X-irradiated L5178Y cells

    International Nuclear Information System (INIS)

    We examined the effect of meta-iodo-benzylguanine (MIBG), an inhibitor of mono (ADP-ribosylation), on growth and clonogenic ability of X-irradiated L5178Y-R (LY-R, radiation resistant) and L5178Y-S (LY-S, radiation sensitive) murine lymphoma cells. We also measured free Ca2+ concentration in LY cells and in human lymphocytes, with the fluorescent calcium chelator, Fura-2. We found no radiosensitization by MIBG in either LY sublines, whereas free Ca2+ concentration increased in MIBG-treated and X-irradiated LY-S cells, as compared to X-irradiated only; in X-irradiated LY-R cells MIGB did not modify the concentration of Ca2+. We conclude that there is no direct relation between the level of free Ca2+, as determined by fluorescent chelator, and the lethal effect of X-irradiation. We also discuss possible artifacts as reasons of incorrect estimation of intracellular Ca2+ concentration. (author). 36 refs, 3 figs, 1 tab

  20. ADP-ribosylation Factor-related Protein 1 Interacts with NS5A and Regulates Hepatitis C Virus Propagation.

    Science.gov (United States)

    Lim, Yun-Sook; Ngo, Huong T T; Lee, Jihye; Son, Kidong; Park, Eun-Mee; Hwang, Soon B

    2016-01-01

    The life cycle of hepatitis C virus (HCV) is tightly coupled to the lipid metabolism of host cells. In order to identify host factors involved in HCV propagation, we have previously screened a small interfering RNA (siRNA) library targeting host genes that control lipid metabolism and lipid droplet (LD) formation using cell culture-grown HCV (HCVcc)-infected cells. In this study, we selected and characterized the gene encoding ADP-ribosylation factor-related protein 1 (ARFRP1). ARFRP1 is essential for LD growth and is involved in the regulation of lipolysis. siRNA-mediated knockdown of ARFRP1 significantly inhibited HCV replication in both subgenomic replicon cells and HCVcc-infected cells. ARFRP1 interacted with NS5A and NS5A partially colocalized with LD. Silencing of ARFRP1 abrogated HCV-induced LD growth and viral protein expressions. Moreover, ARFRP1 recruited synaptosomal-associated protein 23 (SNAP23) to sites in close proximity to LDs in HCV-infected cells. Silencing of ARFRP1 ablated relocalization of SNAP23 to LD. These data indicate that HCV regulates ARFRP1 for LD growth to facilitate viral propagation and thus ARFRP1 may be a potential target for antiviral therapy. PMID:27550144

  1. Molecular cloning, characterization, and expression of human ADP-ribosylation factors: Two guanine nucleotide-dependent activators of cholera toxin

    International Nuclear Information System (INIS)

    ADP-ribosylation factors (ARFs) are small guanine nucleotide-binding proteins that enhance the enzymatic activities of cholera toxin. Two ARF cDNAs, ARF1 and ARF3, were cloned from a human cerebellum library. Based on deduced amino acid sequences and patterns of hybridization of cDNA and oligonucleotide probes with mammalian brain poly(A)+ RNA, human ARF1 is the homologue of bovine ARF1. Human ARF3, which differs from bovine ARF1 and bovine ARF2, appears to represent a newly identified third type of ARF. Hybridization patterns of human ARF cDNA and clone-specific oligonucleotides with poly(A)+ RNA are consistent with the presence of at least two, and perhaps four, separate ARF messages in human brain. In vitro translation of ARF1, ARF2, and ARF3 produced proteins that behaved, by SDS/PAGE, similar to a purified soluble brain ARF. Deduced amino acid sequences of human ARF1 and ARF3 contain regions, similar to those in other G proteins, that are believed to be involved in GTP binding and hydrolysis. ARFS also exhibit a modest degree of homology with a bovine phospholipase C. The observations reported here support the conclusion that the ARFs are members of a multigene family of small guanine nucleotide-binding proteins. Definition of the regulation of ARF mRNAs and of function(s) of recombinant ARF proteins will aid in the elucidation of the physiologic role(s) of ARFs

  2. A presynaptic role for the ADP ribosylation factor (ARF)-specific GDP/GTP exchange factor msec7-1.

    Science.gov (United States)

    Ashery, U; Koch, H; Scheuss, V; Brose, N; Rettig, J

    1999-02-01

    ADP ribosylation factors (ARFs) represent a family of small monomeric G proteins that switch from an inactive, GDP-bound state to an active, GTP-bound state. One member of this family, ARF6, translocates on activation from intracellular compartments to the plasma membrane and has been implicated in regulated exocytosis in neuroendocrine cells. Because GDP release in vivo is rather slow, ARF activation is facilitated by specific guanine nucleotide exchange factors like cytohesin-1 or ARNO. Here we show that msec7-1, a rat homologue of cytohesin-1, translocates ARF6 to the plasma membrane in living cells. Overexpression of msec7-1 leads to an increase in basal synaptic transmission at the Xenopus neuromuscular junction. msec7-1-containing synapses have a 5-fold higher frequency of spontaneous synaptic currents than control synapses. On stimulation, the amplitudes of the resulting evoked postsynaptic currents of msec7-1-overexpressing neurons are increased as well. However, further stimulation leads to a decline in amplitudes approaching the values of control synapses. This transient effect on amplitude is strongly reduced on overexpression of msec7-1E157K, a mutant incapable of translocating ARFs. Our results provide evidence that small G proteins of the ARF family and activating factors like msec7-1 play an important role in synaptic transmission, most likely by making more vesicles available for fusion at the plasma membrane.

  3. Proteome-wide Identification of Poly(ADP-Ribosyl)ation Targets in Different Genotoxic Stress Responses

    DEFF Research Database (Denmark)

    Jungmichel, S.; Rosenthal, F.; Altmeyer, M.;

    2013-01-01

    Poly(ADP-ribos)ylation (PARylation) is a reversible posttranslational modification found in higher eukaryotes. However, little is known about PARylation acceptor proteins. Here, we describe a sensitive proteomics approach based on high-accuracy quantitative mass spectrometry for the identification...... the PARylation of RNA-processing factors THRAP3 and TAF15 under oxidative stress. High-content imaging reveals that PARylation affects the nuclear relocalization of THRAP3 and TAF15, demonstrating the potential of our approach to uncover hitherto unappreciated processes being controlled by specific...

  4. Sam68 Is Required for DNA Damage Responses via Regulating Poly(ADP-ribosyl)ation.

    Science.gov (United States)

    Sun, Xin; Fu, Kai; Hodgson, Andrea; Wier, Eric M; Wen, Matthew G; Kamenyeva, Olena; Xia, Xue; Koo, Lily Y; Wan, Fengyi

    2016-09-01

    The rapid and robust synthesis of polymers of adenosine diphosphate (ADP)-ribose (PAR) chains, primarily catalyzed by poly(ADP-ribose) polymerase 1 (PARP1), is crucial for cellular responses to DNA damage. However, the precise mechanisms through which PARP1 is activated and PAR is robustly synthesized are not fully understood. Here, we identified Src-associated substrate during mitosis of 68 kDa (Sam68) as a novel signaling molecule in DNA damage responses (DDRs). In the absence of Sam68, DNA damage-triggered PAR production and PAR-dependent DNA repair signaling were dramatically diminished. With serial cellular and biochemical assays, we demonstrated that Sam68 is recruited to and significantly overlaps with PARP1 at DNA lesions and that the interaction between Sam68 and PARP1 is crucial for DNA damage-initiated and PARP1-conferred PAR production. Utilizing cell lines and knockout mice, we illustrated that Sam68-deleted cells and animals are hypersensitive to genotoxicity caused by DNA-damaging agents. Together, our findings suggest that Sam68 plays a crucial role in DDR via regulating DNA damage-initiated PAR production. PMID:27635653

  5. mADP-RTs: Versatile virulence factors from bacterial pathogens of plants and mammals

    OpenAIRE

    Lennart eWirthmueller; Banfield, Mark J.

    2012-01-01

    Mono ADP-ribosyltransferases (mADP-RTs) are a family of enzymes that cleave NAD+ and covalently attach the ADP-ribosyl moiety to target proteins. mADP-RTs are well established as important virulence factors of bacteria that infect mammals. Cholera toxin, pertussis toxin and diphteria toxin are three of the best-known examples of mADP-RTs. They modify host target proteins in order to promote infection and/or killing of the host cell. Despite low sequence similarity at the primary amino acid le...

  6. Characterization of two novel ADP ribosylation factors from giant freshwater prawn Macrobrachium rosenbergii and their responses to WSSV challenge.

    Science.gov (United States)

    Ding, Zheng-Feng; Ren, Jie; Tan, Jing-Min; Wang, Zheng; Yin, Shao-Wu; Huang, Ying; Huang, Xin; Wang, Wen; Lan, Jiang-Feng; Ren, Qian

    2015-01-01

    ADP-ribosylation factors (Arfs) are small GTP-binding proteins that have an essential function in intracellular trafficking and organelle structure. To date, little information is available on the Arfs in the economically important giant freshwater prawn Macrobrachium rosenbergii and their relationship to viral infection. Here we identified two Arf genes from M. rosenbergii (MrArf1 and MrArf2) for the first time. Phylogenetic analysis showed that MrArf1, together with MjArf1 from shrimp Marsupenaeus japonicus belonged to Class I Arfs. By contrast, MrArf2 didn't not match any of the Arfs classes of I/II/III, although it could be clustered with an Arf protein from M. japonicas called MjArfn, which may represent an analog of the Arf. MrArf1 was ubiquitously expressed in all the examined tissues, with the highest transcription level in the hepatopancreas, whereas MrArf2 was only highly expressed in the hepatopancreas and exhibited very low levels in the heart, stomach, gills and intestine. The expression level of MrArf1 in the gills was down-regulated post 24 h WSSV challenge, and reached the maximal level at 48 h. MrArf1 in the hepatopancreas went up from 24 to 48 h WSSV challenge. MrArf2 transcript in the gill also went down at 24 h and then was upregulated at 48 h WSSV challenge. MrArf2 increased significantly in the hepatopancreas 24 h after infection and then went down at 48 h WSSV challenge. RNAi results showed that knockdown of MrArf1 or MrArf2 could inhibit the expression of the envelope protein gene vp28 of the WSSV. So, it could be speculated that MrArf1 and MrArf2 might play important roles in the innate immune system against WSSV infection.

  7. Cholera toxin-induced ADP-ribosylation of a 46 kDa protein is decreased in brains of ethanol-fed mice

    International Nuclear Information System (INIS)

    The acute in vitro effects of ethanol on cerebral cortical adenylate cyclase activity and beta-adrenergic receptor characteristics suggested a site of action of ethanol at Gs, the stimulatory guanine nucleotide binding protein. After chronic ethanol ingestion, the beta-adrenergic receptor appeared to be uncoupled (i.e., the form of the receptor with high affinity for agonist was undetectable), and stimulation of adenylate cyclase activity by isoproterenol or guanine nucleotides was reduced, suggesting an alteration in the properties of Gs. To further characterize this change, cholera and pertussis toxin-mediated 32P-ADP-ribosylation of mouse cortical membranes was assessed in mice that had chronically ingested ethanol in a liquid diet. 32P-labeled proteins were separated by SDS-PAGE and quantitated by autoradiography. There was a selective 30-50% decrease in cholera toxin-induced labeling of 46 kDa protein band in membranes of ethanol-fed mice, with no apparent change in pertussis toxin-induced labeling. The 46 kDa protein has a molecular weight similar to that of the alpha subunit of Gs, suggesting a reduced amount of this protein or a change in its characteristics as a substrate for cholera toxin-induced ADP-ribosylation in cortical membranes of ethanol-fed mice

  8. Recovery of human lymphocytes damaged with. gamma. -radiation or enzymatically produced oxygen radicals: different effects of poly(ADP-ribosyl)polymerase inhibitors

    Energy Technology Data Exchange (ETDEWEB)

    Marini, M.; Zunica, G. (Ist. di Istologia ed Embriologia Generale, Bologna (Italy)); Tamba, M. (Consiglio Nazionale delle Ricerche, Bologna (Italy). Lab. di Fotochimica e Radiazioni d' Alta Energia); Cossarizza, A.; Monti, D.; Franceschi, C. (Ist. di Patologia Generale, Modena (Italy))

    1990-08-01

    Quiescent human lymphocytes were damaged in two different ways, both producing toxic oxygen radicals: xanthine oxidase plus hypoxanthine (XOD/HYP), or {gamma}-rays. Under conditions where DNA synthesis was reduced to 10-20% of control, inhibitors of poly(ADP-ribosyl)polymerase (ADPRP, an enzyme that becomes activated in the presence of DNA strand breaks) allowed lymphocytes to recover completely when the damage was caused by XOD/HYP, but they did not affect DNA synthesis of irradiated cells. However, a protective effect of ADPRP inhibitors was observed with irradiated lymphocytes receiving doses {ge}50Gy. Unscheduled DNA synthesis was significantly increased when lymphocytes were damaged by high radiation doses in the presence of ADPRP inhibitors. (author).

  9. ADP-ribosylation factor 1 controls the activation of the phosphatidylinositol 3-kinase pathway to regulate epidermal growth factor-dependent growth and migration of breast cancer cells.

    Science.gov (United States)

    Boulay, Pierre-Luc; Cotton, Mathieu; Melançon, Paul; Claing, Audrey

    2008-12-26

    Activation of intracellular signaling pathways by growth factors is one of the major causes of cancer development and progression. Recent studies have demonstrated that monomeric G proteins of the Ras family are key regulators of cell proliferation, migration, and invasion. Using an invasive breast cancer cell lines, we demonstrate that the ADP-ribosylation factor 1 (ARF1), a small GTPase classically associated with the Golgi, is an important regulator of the biological effects induced by epidermal growth factor. Here, we show that this ARF isoform is activated following epidermal growth factor stimulation and that, in MDA-MB-231 cells, ARF1 is found in dynamic plasma membrane ruffles. Inhibition of endogenous ARF1 expression results in the inhibition of breast cancer cell migration and proliferation. The underlying mechanism involves the activation of the phosphatidylinositol 3-kinase pathway. Our data demonstrate that depletion of ARF1 markedly impairs the recruitment of the phosphatidylinositol 3-kinase catalytic subunit (p110alpha) to the plasma membrane, and the association of the regulatory subunit (p85alpha) to the activated receptor. These results uncover a novel molecular mechanism by which ARF1 regulates breast cancer cell growth and invasion during cancer progression.

  10. The Poly(ADP-ribose) Polymerase Enzyme Tankyrase Antagonizes Activity of the β-Catenin Destruction Complex through ADP-ribosylation of Axin and APC2.

    Science.gov (United States)

    Croy, Heather E; Fuller, Caitlyn N; Giannotti, Jemma; Robinson, Paige; Foley, Andrew V A; Yamulla, Robert J; Cosgriff, Sean; Greaves, Bradford D; von Kleeck, Ryan A; An, Hyun Hyung; Powers, Catherine M; Tran, Julie K; Tocker, Aaron M; Jacob, Kimberly D; Davis, Beckley K; Roberts, David M

    2016-06-10

    Most colon cancer cases are initiated by truncating mutations in the tumor suppressor, adenomatous polyposis coli (APC). APC is a critical negative regulator of the Wnt signaling pathway that participates in a multi-protein "destruction complex" to target the key effector protein β-catenin for ubiquitin-mediated proteolysis. Prior work has established that the poly(ADP-ribose) polymerase (PARP) enzyme Tankyrase (TNKS) antagonizes destruction complex activity by promoting degradation of the scaffold protein Axin, and recent work suggests that TNKS inhibition is a promising cancer therapy. We performed a yeast two-hybrid (Y2H) screen and uncovered TNKS as a putative binding partner of Drosophila APC2, suggesting that TNKS may play multiple roles in destruction complex regulation. We find that TNKS binds a C-terminal RPQPSG motif in Drosophila APC2, and that this motif is conserved in human APC2, but not human APC1. In addition, we find that APC2 can recruit TNKS into the β-catenin destruction complex, placing the APC2/TNKS interaction at the correct intracellular location to regulate β-catenin proteolysis. We further show that TNKS directly PARylates both Drosophila Axin and APC2, but that PARylation does not globally regulate APC2 protein levels as it does for Axin. Moreover, TNKS inhibition in colon cancer cells decreases β-catenin signaling, which we find cannot be explained solely through Axin stabilization. Instead, our findings suggest that TNKS regulates destruction complex activity at the level of both Axin and APC2, providing further mechanistic insight into TNKS inhibition as a potential Wnt pathway cancer therapy. PMID:27068743

  11. Yeast Golgi-localized, γ-Ear–containing, ADP-Ribosylation Factor-binding Proteins Are but Adaptor Protein-1 Is Not Required for Cell-free Transport of Membrane Proteins from the Trans-Golgi Network to the Prevacuolar Compartment

    OpenAIRE

    Abazeed, Mohamed E.; Fuller, Robert S.

    2008-01-01

    Golgi-localized, γ-Ear–containing, ADP-ribosylation factor-binding proteins (GGAs) and adaptor protein-1 (AP-1) mediate clathrin-dependent trafficking of transmembrane proteins between the trans-Golgi network (TGN) and endosomes. In yeast, the vacuolar sorting receptor Vps10p follows a direct pathway from the TGN to the late endosome/prevacuolar compartment (PVC), whereas, the processing protease Kex2p partitions between the direct pathway and an indirect pathway through the early endosome. T...

  12. Increased poly(ADP-ribosyl)ation in skeletal muscle tissue of pediatric patients with severe burn injury: prevention by propranolol treatment

    OpenAIRE

    Oláh, Gábor; Finnerty, Celeste; Sbrana, Elena; Elijah, Itoro; Gerö, Domokos; Herndon, David; Szabó, Csaba

    2011-01-01

    Activation of the nuclear enzyme poly (ADP-ribose) polymerase (PARP) has been shown to promote cellular energetic collapse and cellular necrosis in various forms of critical illness. Most of the evidence implicating the PARP pathway in disease processes is derived from preclinical studies. With respect to PARP and burns, studies in rodent and large animal models of burn injury have demonstrated the activation of PARP in various tissues and the beneficial effect of its pharmacological inhibiti...

  13. A Review of Tandem Mass Spectrometry Characterization of Adenosine Diphosphate-Ribosylated Peptides

    OpenAIRE

    Hengel, Shawna M.; Goodlett, David R.

    2011-01-01

    The use of tandem mass spectrometry to identify and characterize sites of protein adenosine diphosphate (ADP) ribosylation will be reviewed. Specifically, we will focus on data acquisition schemes and fragmentation techniques that provide peptide sequence and modification site information. Also discussed are uses of synthetic standards to aid characterization, and an enzymatic method that converts ADP-ribosylated peptides into ribosyl mono phosphorylated peptides making identification amenabl...

  14. Increased poly(ADP-ribosyl)ation in skeletal muscle tissue of pediatric patients with severe burn injury: prevention by propranolol treatment.

    Science.gov (United States)

    Oláh, Gábor; Finnerty, Celeste C; Sbrana, Elena; Elijah, Itoro; Gerö, Domokos; Herndon, David N; Szabó, Csaba

    2011-07-01

    Activation of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) has been shown to promote cellular energetic collapse and cellular necrosis in various forms of critical illness. Most of the evidence implicating the PARP pathway in disease processes is derived from preclinical studies. With respect to PARP and burns, studies in rodent and large animal models of burn injury have demonstrated the activation of PARP in various tissues and the beneficial effect of its pharmacological inhibition. The aims of the current study were to measure the activation of PARP in human skeletal muscle biopsies at various stages of severe pediatric burn injury and to identify the cell types where this activation may occur. Another aim of the study was to test the effect of propranolol (an effective treatment of patients with burns) on the activation of PARP in skeletal muscle biopsies. Poly(ADP-ribose) polymerase activation was measured by Western blotting for its product, poly(ADP-ribose) (PAR). The localization of PARP activation was determined by PAR immunohistochemistry. The results showed that PARP becomes activated in the skeletal muscle tissue after burns, with the peak of the activation occurring in the middle stage of the disease (13-18 days after burns). Even at the late stage of the disease (69-369 days after burn), an elevated degree of PARP activation persisted in some of the patients. Immunohistochemical studies localized the staining of PAR primarily to vascular endothelial cells and occasionally to resident mononuclear cells. There was a marked suppression of PARP activation in the skeletal muscle biopsies of patients who received propranolol treatment. We conclude that human burn injury is associated with the activation of PARP. We hypothesize that this response may contribute to the inflammatory responses and cell dysfunction in burns. Some of the clinical benefit of propranolol in burns may be related to its inhibitory effect on PARP activation.

  15. Poly(ADP-ribosyl)ation links the chromatin remodeler SMARCA5/SNF2H to RNF168-dependent DNA damage signaling

    DEFF Research Database (Denmark)

    Smeenk, G.; Wiegant, W.W.; Luijsterburg, M.S.;

    2013-01-01

    Ionizing radiation (IR)-induced DNA double-strand breaks (DSBs) arising in native chromatin elicit an RNF8/RNF168-dependent ubiquitylation response, which triggers the recruitment of various repair factors. Precisely how this response is regulated in the context of chromatin remains largely...... unexplored. Here, we show that SMARCA5/SNF2H, the catalytic subunit of ISWI chromatin remodeling complexes, is recruited to DSBs in a poly(ADP-ribose) polymerase 1 (PARP1)-dependent manner. Remarkably, PARP activity, although dispensable for the efficient spreading of νH2AX into damaged chromatin......, selectively promotes spreading of SMARCA5, the E3 ubiquitin ligase RNF168, ubiquitin conjugates and the ubiquitin-binding factors RAD18 and the RAP80-BRCA1 complex throughout DSB-flanking chromatin. This suggests that PARP regulates the spatial organization of the RNF168-driven ubiquitin response to DNA...

  16. Increased poly(ADP-ribosyl)ation in skeletal muscle tissue of pediatric patients with severe burn injury: prevention by propranolol treatment

    Science.gov (United States)

    Oláh, Gábor; Finnerty, Celeste; Sbrana, Elena; Elijah, Itoro; Gerö, Domokos; Herndon, David; Szabó, Csaba

    2011-01-01

    Summary Activation of the nuclear enzyme poly (ADP-ribose) polymerase (PARP) has been shown to promote cellular energetic collapse and cellular necrosis in various forms of critical illness. Most of the evidence implicating the PARP pathway in disease processes is derived from preclinical studies. With respect to PARP and burns, studies in rodent and large animal models of burn injury have demonstrated the activation of PARP in various tissues and the beneficial effect of its pharmacological inhibition. The aim of the current study was to measure the activation of PARP in human skeletal muscle biopsies at various stages of severe pediatric burn injury and to identify the cell types where this activation may occur. Another aim of the study was to test the effect of propranolol (an effective treatment of patients with burns), on the activation of PARP in skeletal muscle biopsies. PARP activation was measured by Western blotting for its product, poly(ADP-ribose) (PAR). The localization of PARP activation was determined by PAR immunohistochemistry. The results showed that PARP becomes activated in the skeletal muscle tissue after burns, with the peak of the activation occurring in the middle stage of the disease (13–18 days after burns). Even at the late stage of the disease (69–369 days post-burn) an elevated degree of PARP activation persisted in some of the patients. Immunohistochemical studies localized the staining of PAR primarily to vascular endothelial cells, and occasionally to resident mononuclear cells. There was a marked suppression of PARP activation in the skeletal muscle biopsies of patients who received propranolol treatment. We conclude that human burn injury is associated with the activation of PARP. We hypothesize that this response may contribute to the inflammatory responses and cell dysfunction in burns. Some of the clinical benefit of propranolol in burns may be related to its inhibitory effect on PARP activation. PMID:21368715

  17. Novel cholix toxin variants, ADP-ribosylating toxins in Vibrio cholerae non-O1/non-O139 strains, and their pathogenicity.

    Science.gov (United States)

    Awasthi, Sharda Prasad; Asakura, Masahiro; Chowdhury, Nityananda; Neogi, Sucharit Basu; Hinenoya, Atsushi; Golbar, Hossain M; Yamate, Jyoji; Arakawa, Eiji; Tada, Toshiji; Ramamurthy, T; Yamasaki, Shinji

    2013-02-01

    Cholix toxin (ChxA) is a recently discovered exotoxin in Vibrio cholerae which has been characterized as a third member of the eukaryotic elongation factor 2-specific ADP-ribosyltransferase toxins, in addition to exotoxin A of Pseudomonas aeruginosa and diphtheria toxin of Corynebacterium diphtheriae. These toxins consist of three characteristic domains for receptor binding, translocation, and catalysis. However, there is little information about the prevalence of chxA and its genetic variations and pathogenic mechanisms. In this study, we screened the chxA gene in a large number (n = 765) of V. cholerae strains and observed its presence exclusively in non-O1/non-O139 strains (27.0%; 53 of 196) and not in O1 (n = 485) or O139 (n = 84). Sequencing of these 53 chxA genes generated 29 subtypes which were grouped into three clusters designated chxA I, chxA II, and chxA III. chxA I belongs to the prototype, while chxA II and chxA III are newly discovered variants. ChxA II and ChxA III had unique receptor binding and catalytic domains, respectively, in comparison to ChxA I. Recombinant ChxA I (rChxA I) and rChxA II but not rChxA III showed variable cytotoxic effects on different eukaryotic cells. Although rChxA II was more lethal to mice than rChxA I when injected intravenously, no enterotoxicity of any rChxA was observed in a rabbit ileal loop test. Hepatocytes showed coagulation necrosis in rChxA I- or rChxA II-treated mice, seemingly the major target for ChxA. The present study illustrates the potential of ChxA as an important virulence factor in non-O1/non-O139 V. cholerae, which may be associated with extraintestinal infections rather than enterotoxicity.

  18. Rho-modifying bacterial protein toxins.

    Science.gov (United States)

    Aktories, Klaus

    2015-12-01

    Rho proteins are targets of numerous bacterial protein toxins, which manipulate the GTP-binding proteins by covalent modifications, including ADP ribosylation, glycosylation, adenylylation, proteolytic cleavage and deamidation. Bacterial toxins are important virulence factors but are also potent and efficient pharmacological tools to study the physiological functions of their eukaryotic targets. Recent studies indicate that amazing variations exist in the molecular mechanisms by which toxins attack Rho proteins, which are discussed here.

  19. Changes in NAD/ADP-ribose metabolism in rectal cancer

    Directory of Open Access Journals (Sweden)

    L. Yalcintepe

    2005-03-01

    Full Text Available The extent of ADP-ribosylation in rectal cancer was compared to that of the corresponding normal rectal tissue. Twenty rectal tissue fragments were collected during surgery from patients diagnosed as having rectal cancer on the basis of pathology results. The levels of ADP-ribosylation in rectum cancer tissue samples (95.9 ± 22.1 nmol/ml was significantly higher than in normal tissues (11.4 ± 4 nmol/ml. The level of NAD+ glycohydrolase and ADP-ribosyl cyclase activities in rectal cancer and normal tissue samples were measured. Cancer tissues had significantly higher NAD+ glycohydrolase and ADP-ribosyl cyclase activities than the control tissues (43.3 ± 9.1 vs 29.2 ± 5.2 and 6.2 ± 1.6 vs 1.6 ± 0.4 nmol mg-1 min-1. Approximately 75% of the NAD+ concentration was consumed as substrate in rectal cancer, with changes in NAD+/ADP-ribose metabolism being observed. When [14C]-ADP-ribosylated tissue samples were subjected to SDS-PAGE, autoradiographic analysis revealed that several proteins were ADP-ribosylated in rectum tissue. Notably, the radiolabeling of a 113-kDa protein was remarkably greater than that in control tissues. Poly(ADP-ribosylation of the 113-kDa protein in rectum cancer tissues might be enhanced with its proliferative activity, and poly(ADP-ribosylation of the same protein in rectum cancer patients might be an indicator of tumor diagnosis.

  20. PARP16/ARTD15 is a novel endoplasmic-reticulum-associated mono-ADP-ribosyltransferase that interacts with, and modifies karyopherin-ß1.

    Directory of Open Access Journals (Sweden)

    Simone Di Paola

    Full Text Available BACKGROUND: Protein mono-ADP-ribosylation is a reversible post-translational modification that modulates the function of target proteins. The enzymes that catalyze this reaction in mammalian cells are either bacterial pathogenic toxins or endogenous cellular ADP-ribosyltransferases. The latter include members of three different families of proteins: the well characterized arginine-specific ecto-enzymes ARTCs, two sirtuins and, more recently, novel members of the poly(ADP-ribose polymerase (PARP/ARTD family that have been suggested to act as cellular mono-ADP-ribosyltransferases. Here, we report on the characterisation of human ARTD15, the only known ARTD family member with a putative C-terminal transmembrane domain. METHODOLOGY/PRINCIPAL FINDINGS: Immunofluorescence and electron microscopy were performed to characterise the sub-cellular localisation of ARTD15, which was found to be associated with membranes of the nuclear envelope and endoplasmic reticulum. The orientation of ARTD15 was determined using protease protection assay, and is shown to be a tail-anchored protein with a cytosolic catalytic domain. Importantly, by combining immunoprecipitation with mass spectrometry and using cell lysates from cells over-expressing FLAG-ARTD15, we have identified karyopherin-ß1, a component of the nuclear trafficking machinery, as a molecular partner of ARTD15. Finally, we demonstrate that ARTD15 is a mono-ADP-ribosyltransferase able to induce the ADP-ribosylation of karyopherin-ß1, thus defining the first substrate for this enzyme. CONCLUSIONS/SIGNIFICANCE: Our data reveal that ARTD15 is a novel ADP-ribosyltransferase enzyme with a new intracellular location. Finally, the identification of karyopherin-ß1 as a target of ARTD15-mediated ADP-ribosylation, hints at a novel regulatory mechanism of karyopherin-ß1 functions.

  1. The effect of ADP-ribosylation factor antagonist on alkali-burn induced corneal neovascularization%ADP-核糖基化因子抑制剂在实验性碱烧伤诱导角膜新生血管中的作用及其机制

    Institute of Scientific and Technical Information of China (English)

    刘高勤; 吴敬; 陈志刚; 肖艳辉; 陆培荣

    2014-01-01

    照组为0.72±0.11,2个组间差异有统计学意义(t=-3.87,P<0.05).全角膜铺片免疫荧光组织化学法检测表明,ARF抑制剂干预组小鼠角膜组织CD31表达面积明显小于PBS对照组.造模后14 d,Western blot法检测显示,ARF抑制剂干预组VEGF蛋白表达水平为1.20±0.21,明显低于PBS对照组的2.47±0.33,差异有统计学意义(t=-5.62,P<0.05).CCK8法检测结果显示,随着ARF抑制剂质量浓度的增加,ARF抑制剂对人RECs的抑制率增加,差异有统计学意义(F=8.47,P=0.02).细胞划痕实验后24 h,100μg/L和1 000 μg/L ARF抑制剂干预组人RECs迁移距离分别为(5.46±1.32) μm和(5.04±1.68)μm,与PBS对照组的(8.49±1.18) μm相比明显减小,差异均有统计学意义(t=-2.94、-2.91,P<0.05). 结论 ARF抑制剂能抑制碱烧伤诱导的CNV发生,可能与其下调VEGF的表达以及抑制血管内皮细胞的增生和迁移有关.%Background Corneal neovascularization (CNV) is one of the causes of corneal blindness.Studies showed that ADP-ribosylation factor (ARF) can regulate the growth of tumor cells,and inhibiting ARF will decrease angiogenesis.However,whether ARF antagonist plays an action on CNV is unclear.Objective The aim of this study was to explore the effect and mechanism of ARF inhibitor on alkali-burn induced CNV.Methods Sixty clean male BABL/c mice aged 7-8 weeks were divided into PBS group and ARF antagonist group according to randomized number table.CNV models were induced by NaOH burn method in all the mice.ARF at the concentration of 0.5 g/L(0.5 ml) was intraperitoneally injected 3 times per week for 1 week followed the induction of CNV in the ARF antagonist group,and 0.5 ml PBS was used in the PBS group.CNV was examined 2,4,7,14 days after injection by the slit lamp microscope and the CNV related area in the cornea was calculated.Betore modeling(0 day) and 4,7,14 days after modeling,real-time PCR and Western blot were used to analyze the expressions of ARF mRNA and protein in the

  2. Yeast Golgi-localized, gamma-Ear-containing, ADP-ribosylation factor-binding proteins are but adaptor protein-1 is not required for cell-free transport of membrane proteins from the trans-Golgi network to the prevacuolar compartment.

    Science.gov (United States)

    Abazeed, Mohamed E; Fuller, Robert S

    2008-11-01

    Golgi-localized, gamma-Ear-containing, ADP-ribosylation factor-binding proteins (GGAs) and adaptor protein-1 (AP-1) mediate clathrin-dependent trafficking of transmembrane proteins between the trans-Golgi network (TGN) and endosomes. In yeast, the vacuolar sorting receptor Vps10p follows a direct pathway from the TGN to the late endosome/prevacuolar compartment (PVC), whereas, the processing protease Kex2p partitions between the direct pathway and an indirect pathway through the early endosome. To examine the roles of the Ggas and AP-1 in TGN-PVC transport, we used a cell-free assay that measures delivery to the PVC of either Kex2p or a chimeric protein (K-V), in which the Vps10p cytosolic tail replaces the Kex2p tail. Either antibody inhibition or dominant-negative Gga2p completely blocked K-V transport but only partially blocked Kex2p transport. Deletion of APL2, encoding the beta subunit of AP-1, did not affect K-V transport but partially blocked Kex2p transport. Residual Kex2p transport seen with apl2Delta membranes was insensitive to dominant-negative Gga2p, suggesting that the apl2Delta mutation causes Kex2p to localize to a compartment that precludes Gga-dependent trafficking. These results suggest that yeast Ggas facilitate the specific and direct delivery of Vps10p and Kex2p from the TGN to the PVC and that AP-1 modulates Kex2p trafficking through a distinct pathway, presumably involving the early endosome. PMID:18784256

  3. Yeast Golgi-localized, γ-Ear–containing, ADP-Ribosylation Factor-binding Proteins Are but Adaptor Protein-1 Is Not Required for Cell-free Transport of Membrane Proteins from the Trans-Golgi Network to the Prevacuolar Compartment

    Science.gov (United States)

    Abazeed, Mohamed E.

    2008-01-01

    Golgi-localized, γ-Ear–containing, ADP-ribosylation factor-binding proteins (GGAs) and adaptor protein-1 (AP-1) mediate clathrin-dependent trafficking of transmembrane proteins between the trans-Golgi network (TGN) and endosomes. In yeast, the vacuolar sorting receptor Vps10p follows a direct pathway from the TGN to the late endosome/prevacuolar compartment (PVC), whereas, the processing protease Kex2p partitions between the direct pathway and an indirect pathway through the early endosome. To examine the roles of the Ggas and AP-1 in TGN–PVC transport, we used a cell-free assay that measures delivery to the PVC of either Kex2p or a chimeric protein (K-V), in which the Vps10p cytosolic tail replaces the Kex2p tail. Either antibody inhibition or dominant-negative Gga2p completely blocked K-V transport but only partially blocked Kex2p transport. Deletion of APL2, encoding the β subunit of AP-1, did not affect K-V transport but partially blocked Kex2p transport. Residual Kex2p transport seen with apl2Δ membranes was insensitive to dominant-negative Gga2p, suggesting that the apl2Δ mutation causes Kex2p to localize to a compartment that precludes Gga-dependent trafficking. These results suggest that yeast Ggas facilitate the specific and direct delivery of Vps10p and Kex2p from the TGN to the PVC and that AP-1 modulates Kex2p trafficking through a distinct pathway, presumably involving the early endosome. PMID:18784256

  4. Identification of an enzymatic activity that hydrolyzes protein-bound ADP-ribose in skeletal muscle.

    Science.gov (United States)

    Chang, Y C; Soman, G; Graves, D J

    1986-09-30

    An enzymatic activity present in high-speed supernatant fluids of rat skeletal muscle was found that catalyzes the release of ADP-ribose from ADP-ribosylated-modified lysozyme. The nature of the product was proved by chromatographic studies and proton nuclear magnetic resonance spectroscopy. The enzyme activity is stimulated by Mg2+, dithioerythritol, and flouride. These results and those published earlier (Soman, G., Mickelson, J.R., Louis, C.F., and Graves, D.J. (1984) Biochem. Biophys. Res. Commun. 120, 973-980) show that ADP-ribosylation is a reversible process in skeletal muscle.

  5. Poly(ADP-Ribose) Polymerase 1 Promotes Oxidative-Stress-Induced Liver Cell Death via Suppressing Farnesoid X Receptor α

    OpenAIRE

    Wang, Cheng; Zhang, Fengxiao; Wang, Lin; Zhang, Yanqing; Li, Xiangrao; Huang, Kun; Du, Meng; Liu, Fangmei; Huang, Shizheng; Guan, Youfei; Huang, Dan; Huang, Kai

    2013-01-01

    Farnesoid X receptor α (FXR) is highly expressed in the liver and regulates the expression of various genes involved in liver repair. In this study, we demonstrated that activated poly(ADP-ribose) polymerase 1 (PARP1) promoted hepatic cell death by inhibiting the expression of FXR-dependent hepatoprotective genes. PARP1 could bind to and poly(ADP-ribosyl)ate FXR. Poly(ADP-ribosyl)ation dissociated FXR from the FXR response element (FXRE), present in the promoters of target genes, and suppress...

  6. Differential and Concordant Roles for Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribose) in Regulating WRN and RECQL5 Activities

    OpenAIRE

    Khadka, Prabhat; Hsu, Joseph K; Veith, Sebastian; Tadokoro, Takashi; Shamanna, Raghavendra A.; Mangerich, Aswin; Croteau, Deborah L.; Bohr, Vilhelm A.

    2015-01-01

    Poly(ADP-ribose) (PAR) polymerase 1 (PARP1) catalyzes the poly(ADP-ribosyl)ation (PARylation) of proteins, a posttranslational modification which forms the nucleic acid-like polymer PAR. PARP1 and PAR are integral players in the early DNA damage response, since PARylation orchestrates the recruitment of repair proteins to sites of damage. Human RecQ helicases are DNA unwinding proteins that are critical responders to DNA damage, but how their recruitment and activities are regulated by PARPs ...

  7. Fine-tuning of Smad protein function by poly(ADP-ribose polymerases and poly(ADP-ribose glycohydrolase during transforming growth factor β signaling.

    Directory of Open Access Journals (Sweden)

    Markus Dahl

    Full Text Available BACKGROUND: Initiation, amplitude, duration and termination of transforming growth factor β (TGFβ signaling via Smad proteins is regulated by post-translational modifications, including phosphorylation, ubiquitination and acetylation. We previously reported that ADP-ribosylation of Smads by poly(ADP-ribose polymerase 1 (PARP-1 negatively influences Smad-mediated transcription. PARP-1 is known to functionally interact with PARP-2 in the nucleus and the enzyme poly(ADP-ribose glycohydrolase (PARG can remove poly(ADP-ribose chains from target proteins. Here we aimed at analyzing possible cooperation between PARP-1, PARP-2 and PARG in regulation of TGFβ signaling. METHODS: A robust cell model of TGFβ signaling, i.e. human HaCaT keratinocytes, was used. Endogenous Smad3 ADP-ribosylation and protein complexes between Smads and PARPs were studied using proximity ligation assays and co-immunoprecipitation assays, which were complemented by in vitro ADP-ribosylation assays using recombinant proteins. Real-time RT-PCR analysis of mRNA levels and promoter-reporter assays provided quantitative analysis of gene expression in response to TGFβ stimulation and after genetic perturbations of PARP-1/-2 and PARG based on RNA interference. RESULTS: TGFβ signaling rapidly induces nuclear ADP-ribosylation of Smad3 that coincides with a relative enhancement of nuclear complexes of Smads with PARP-1 and PARP-2. Inversely, PARG interacts with Smads and can de-ADP-ribosylate Smad3 in vitro. PARP-1 and PARP-2 also form complexes with each other, and Smads interact and activate auto-ADP-ribosylation of both PARP-1 and PARP-2. PARP-2, similar to PARP-1, negatively regulates specific TGFβ target genes (fibronectin, Smad7 and Smad transcriptional responses, and PARG positively regulates these genes. Accordingly, inhibition of TGFβ-mediated transcription caused by silencing endogenous PARG expression could be relieved after simultaneous depletion of PARP-1. CONCLUSION

  8. Exchange of glutamine-217 to glutamate of Clostridium limosum exoenzyme C3 turns the asparagine-specific ADP-ribosyltransferase into an arginine-modifying enzyme.

    Science.gov (United States)

    Vogelsgesang, Martin; Aktories, Klaus

    2006-01-24

    C3-like ADP-ribosyltransferaseses are produced by Clostridium species, Bacillus cereus, and various Staphylococcus aureus strains. The exoenzymes modify the low-molecular-mass GTPases RhoA, B, and C. In structural studies of C3-like exoenzymes, an ARTT-motif (ADP-ribosylating turn-turn motif) was identified that appears to be involved in substrate specificity and recognition (Han, S., Arvai, A. S., Clancy, S. B., Tainer, J. A. (2001) J. Mol. Biol. 305, 95-107). Exchange of Gln217, which is a key residue of the ARTT-motif, to Glu in C3 from Clostridium limosum results in inhibition of ADP-ribosyltransferase activity toward RhoA. The mutant protein is still capable of NAD-binding and possesses NAD+ glycohydrolase activity. Whereas recombinant wild-type C3 modifies Rho proteins specifically at an asparagine residue (Asn41), Gln217Glu-C3 is capable of ADP-ribosylation of poly-arginine but not poly-asparagine. Soybean trypsin inhibitor, a model substrate for many arginine-specific ADP-ribosyltransferases, is modified by the Gln217Glu-C3 transferase. Also in C3 ADP-ribosyltransferases from Clostridium botulinum and B. cereus, the exchange of the equivalent Gln residue to Glu blocked asparagine modification of RhoA but elicited arginine-specific ADP-ribosylation. Moreover, the Gln217Glu-C3lim transferase was able to ADP-ribosylate recombinant wild-type C3lim at Arg86, resulting in decrease in ADP-ribosyltransferase activity of the wild-type enzyme. The data indicate that the exchange of one amino acid residue in the ARTT-motif turns the asparagine-modifying ADP-ribosyltransferases of the C3 family into arginine-ADP-ribosylating transferases.

  9. Vault-poly-ADP-ribose polymerase in the Octopus vulgaris brain: a regulatory factor of actin polymerization dynamic.

    Science.gov (United States)

    De Maio, Anna; Natale, Emiliana; Rotondo, Sergio; Di Cosmo, Anna; Faraone-Mennella, Maria Rosaria

    2013-09-01

    Our previous behavioural, biochemical and immunohistochemical analyses conducted in selected regions (supra/sub oesophageal masses) of the Octopus vulgaris brain detected a cytoplasmic poly-ADP-ribose polymerase (more than 90% of total enzyme activity). The protein was identified as the vault-free form of vault-poly-ADP-ribose polymerase. The present research extends and integrates the biochemical characterization of poly-ADP-ribosylation system, namely, reaction product, i.e., poly-ADP-ribose, and acceptor proteins, in the O. vulgaris brain. Immunochemical analyses evidenced that the sole poly-ADP-ribose acceptor was the octopus cytoskeleton 50-kDa actin. It was present in both free, endogenously poly-ADP-ribosylated form (70kDa) and in complex with V-poly-ADP-ribose polymerase and poly-ADP-ribose (260kDa). The components of this complex, alkali and high salt sensitive, were purified and characterized. The kind and the length of poly-ADP-ribose corresponded to linear chains of 30-35 ADP-ribose units, in accordance with the features of the polymer synthesized by the known vault-poly-ADP-ribose polymerase. In vitro experiments showed that V-poly-ADP-ribose polymerase activity of brain cytoplasmic fraction containing endogenous actin increased upon the addition of commercial actin and was highly reduced by ATP. Anti-actin immunoblot of the mixture in the presence and absence of ATP showed that the poly-ADP-ribosylation of octopus actin is a dynamic process balanced by the ATP-dependent polymerization of the cytoskeleton protein, a fundamental mechanism for synaptic plasticity.

  10. The Neisseria meningitidis ADP-Ribosyltransferase NarE Enters Human Epithelial Cells and Disrupts Epithelial Monolayer Integrity.

    Directory of Open Access Journals (Sweden)

    Maria Valeri

    Full Text Available Many pathogenic bacteria utilize ADP-ribosylating toxins to modify and impair essential functions of eukaryotic cells. It has been previously reported that Neisseria meningitidis possesses an ADP-ribosyltransferase enzyme, NarE, retaining the capacity to hydrolyse NAD and to transfer ADP-ribose moiety to arginine residues in target acceptor proteins. Here we show that upon internalization into human epithelial cells, NarE gains access to the cytoplasm and, through its ADP-ribosylating activity, targets host cell proteins. Notably, we observed that these events trigger the disruption of the epithelial monolayer integrity and the activation of the apoptotic pathway. Overall, our findings provide, for the first time, evidence for a biological activity of NarE on host cells, suggesting its possible involvement in Neisseria pathogenesis.

  11. Overexpression, purification, and partial characterization of ADP-ribosyltransferases modA and modB of bacteriophage T4.

    Science.gov (United States)

    Tiemann, B; Depping, R; Rüger, W

    1999-01-01

    There is increasing experimental evidence that ADP-ribosylation of host proteins is an important means to regulate gene expression of bacteriophage T4. Surprisingly, this phage codes for three different ADP-ribosyltransferases, gene products Alt, ModA, and ModB, modifying partially overlapping sets of host proteins. While gene product Alt already has been isolated as a recombinant protein and its action on host RNA polymerases and transcription regulation have been studied, the nucleotide sequences of the two mod genes was published only recently. Their mode of action in the course of the infection cycle and the consequences of the ADP-ribosylations catalyzed by these enzymes remain to be investigated. Here we describe the cloning of the genes, the overexpression, purification, and partial characterization of ADP-ribosyltransferases ModA and ModB. Both proteins seem to act independently, and the ADP-ribosyl moieties are transferred to different sets of host proteins. While gene product ModA, similarly to the Alt protein, acts also on the alpha-subunit of host RNA polymerase, the ModB activity serves another set of proteins, one of which was identified as the S1 protein associated with the 30S subunit of the E. coli ribosomes.

  12. PARP2 Is the Predominant Poly(ADP-Ribose Polymerase in Arabidopsis DNA Damage and Immune Responses.

    Directory of Open Access Journals (Sweden)

    Junqi Song

    2015-05-01

    Full Text Available Poly (ADP-ribose polymerases (PARPs catalyze the transfer of multiple poly(ADP-ribose units onto target proteins. Poly(ADP-ribosylation plays a crucial role in a variety of cellular processes including, most prominently, auto-activation of PARP at sites of DNA breaks to activate DNA repair processes. In humans, PARP1 (the founding and most characterized member of the PARP family accounts for more than 90% of overall cellular PARP activity in response to DNA damage. We have found that, in contrast with animals, in Arabidopsis thaliana PARP2 (At4g02390, rather than PARP1 (At2g31320, makes the greatest contribution to PARP activity and organismal viability in response to genotoxic stresses caused by bleomycin, mitomycin C or gamma-radiation. Plant PARP2 proteins carry SAP DNA binding motifs rather than the zinc finger domains common in plant and animal PARP1 proteins. PARP2 also makes stronger contributions than PARP1 to plant immune responses including restriction of pathogenic Pseudomonas syringae pv. tomato growth and reduction of infection-associated DNA double-strand break abundance. For poly(ADP-ribose glycohydrolase (PARG enzymes, we find that Arabidopsis PARG1 and not PARG2 is the major contributor to poly(ADP-ribose removal from acceptor proteins. The activity or abundance of PARP2 is influenced by PARP1 and PARG1. PARP2 and PARP1 physically interact with each other, and with PARG1 and PARG2, suggesting relatively direct regulatory interactions among these mediators of the balance of poly(ADP-ribosylation. As with plant PARP2, plant PARG proteins are also structurally distinct from their animal counterparts. Hence core aspects of plant poly(ADP-ribosylation are mediated by substantially different enzymes than in animals, suggesting the likelihood of substantial differences in regulation.

  13. Discovery of novel poly(ADP-ribose) glycohydrolase inhibitors by a quantitative assay system using dot-blot with anti-poly(ADP-ribose)

    Energy Technology Data Exchange (ETDEWEB)

    Okita, Naoyuki, E-mail: nokita7@rs.noda.tus.ac.jp [Genome and Drug Research Center, Tokyo University of Science (Japan); Ashizawa, Daisuke; Ohta, Ryo [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-0022 (Japan); Abe, Hideaki [Genome and Drug Research Center, Tokyo University of Science (Japan); Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-0022 (Japan); Tanuma, Sei-ichi, E-mail: tanuma@rs.noda.tus.ac.jp [Genome and Drug Research Center, Tokyo University of Science (Japan); Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-0022 (Japan)

    2010-02-19

    Poly(ADP-ribosyl)ation, which is mainly regulated by poly(ADP-ribose) polymerase (PARP) and poly(ADP-ribose) glycohydrolase (PARG), is a unique protein modification involved in cellular responses such as DNA repair and replication. PARG hydrolyzes glycosidic linkages of poly(ADP-ribose) synthesized by PARP and liberates ADP-ribose residues. Recent studies have suggested that inhibitors of PARG are able to be potent anti-cancer drug. In order to discover the potent and specific Inhibitors of PARG, a quantitative and high-throughput screening assay system is required. However, previous PARG assay systems are not appropriate for high-throughput screening because PARG activity is measured by radioactivities of ADP-ribose residues released from radioisotope (RI)-labeled poly(ADP-ribose). In this study, we developed a non-RI and quantitative assay system for PARG activity based on dot-blot assay using anti-poly(ADP-ribose) and nitrocellulose membrane. By our method, the maximum velocity (V{sub max}) and the michaelis constant (k{sub m}) of PARG reaction were 4.46 {mu}M and 128.33 {mu}mol/min/mg, respectively. Furthermore, the IC50 of adenosine diphosphate (hydroxymethyl) pyrrolidinediol (ADP-HPD), known as a non-competitive PARG inhibitor, was 0.66 {mu}M. These kinetics values were similar to those obtained by traditional PARG assays. By using our assay system, we discovered two novel PARG inhibitors that have xanthene scaffold. Thus, our quantitative and convenient method is useful for a high-throughput screening of PARG specific inhibitors.

  14. ADP-ribosylation factor 6 mediates E-cadherin recovery by chemical chaperones.

    Directory of Open Access Journals (Sweden)

    Joana Figueiredo

    Full Text Available E-cadherin plays a powerful tumor suppressor role. Germline E-cadherin mutations justify 30% of Hereditary Diffuse Gastric Cancer (HDGC and missense mutations are found in 30% of these families. We found possible to restore in vitro mutant E-cadherin associated to HDGC syndrome by using Chemical Chaperones (CCs. Herein, our aim was to disclose the molecular mechanisms underlying the CCs effects in E-cadherin regulation. Using cells stably expressing WT E-cadherin or two HDGC-associated missense mutations, we show that upon DMSO treatment, not only mutant E-cadherin is restored and stabilized at the plasma membrane (PM, but also Arf6 and PIPKIγ expressions are altered. We show that modulation of Arf6 expression partially mimics the effect of CCs, suggesting that the cellular effects observed upon CCs treatment are mediated by Arf6. Further, we show that E-cadherin expression recovery is specifically linked to Arf6 due to its role on endocytosis and recycling pathways. Finally, we demonstrated that, as DMSO, several others CCs are able to modulate the trafficking machinery through an Arf6 dependent mechanism. Interestingly, the more effective compounds in E-cadherin recovery to PM are those that simultaneously inhibit Arf6 and stimulate PIPKIγ expression and binding to E-cadherin. Here, we present the first evidence of a direct influence of CCs in cellular trafficking machinery and we show that this effect is of crucial importance in the context of juxtamembrane E-cadherin missense mutations associated to HDGC. We propose that this influence should be taken into account when exploring the therapeutic potential of this type of chemicals in genetic diseases associated to protein-misfolding.

  15. ADP's ABCs of Training

    Science.gov (United States)

    Weinstein, Margery

    2010-01-01

    When a company's core competence is processing data, it is sometimes easy to lose sight of the obvious--the information right under its nose. In the case of Automatic Data Processing, Inc. (ADP), a business outsourcing company specializing in human resources, payroll, tax, and benefits administrations solutions, that is not a problem. Through…

  16. ModA and ModB, two ADP-ribosyltransferases encoded by bacteriophage T4: catalytic properties and mutation analysis.

    Science.gov (United States)

    Tiemann, Bernd; Depping, Reinhard; Gineikiene, Egle; Kaliniene, Laura; Nivinskas, Rimas; Rüger, Wolfgang

    2004-11-01

    Bacteriophage T4 encodes three ADP-ribosyltransferases, Alt, ModA, and ModB. These enzymes participate in the regulation of the T4 replication cycle by ADP-ribosylating a defined set of host proteins. In order to obtain a better understanding of the phage-host interactions and their consequences for regulating the T4 replication cycle, we studied cloning, overexpression, and characterization of purified ModA and ModB enzymes. Site-directed mutagenesis confirmed that amino acids, as deduced from secondary structure alignments, are indeed decisive for the activity of the enzymes, implying that the transfer reaction follows the Sn1-type reaction scheme proposed for this class of enzymes. In vitro transcription assays performed with Alt- and ModA-modified RNA polymerases demonstrated that the Alt-ribosylated polymerase enhances transcription from T4 early promoters on a T4 DNA template, whereas the transcriptional activity of ModA-modified polymerase, without the participation of T4-encoded auxiliary proteins for middle mode or late transcription, is reduced. The results presented here support the conclusion that ADP-ribosylation of RNA polymerase and of other host proteins allows initial phage-directed mRNA synthesis reactions to escape from host control. In contrast, subsequent modification of the other cellular target proteins limits transcription from phage early genes and participates in redirecting transcription to phage middle and late genes.

  17. Structural and biochemical characterization of NAR E, an iron containing ADP-ribosyltransferase from neisseria meningitidis

    NARCIS (Netherlands)

    Koehler, C.; Carlier, L.P.A.; Veggi, D.; Balducci, C.; Di Marcello, F.; Ferrer-Navarro, M.; Pizza, M.; Daura, X.; Soriani, M.; Boelens, R.; Bonvin, A.M.J.J.

    2011-01-01

    NarE is a 16 kDa protein identified from Neisseria meningitidis, one of the bacterial pathogens responsible for meningitis. NarE belongs to the family of ADP-ribosyltransferases (ADPRT) and catalyzes the transfer of ADP-ribose moieties to arginine residues in target protein acceptors. Many pathogeni

  18. The XRCC1 phosphate-binding pocket binds poly (ADP-ribose) and is required for XRCC1 function.

    Science.gov (United States)

    Breslin, Claire; Hornyak, Peter; Ridley, Andrew; Rulten, Stuart L; Hanzlikova, Hana; Oliver, Antony W; Caldecott, Keith W

    2015-08-18

    Poly (ADP-ribose) is synthesized at DNA single-strand breaks and can promote the recruitment of the scaffold protein, XRCC1. However, the mechanism and importance of this process has been challenged. To address this issue, we have characterized the mechanism of poly (ADP-ribose) binding by XRCC1 and examined its importance for XRCC1 function. We show that the phosphate-binding pocket in the central BRCT1 domain of XRCC1 is required for selective binding to poly (ADP-ribose) at low levels of ADP-ribosylation, and promotes interaction with cellular PARP1. We also show that the phosphate-binding pocket is required for EGFP-XRCC1 accumulation at DNA damage induced by UVA laser, H2O2, and at sites of sub-nuclear PCNA foci, suggesting that poly (ADP-ribose) promotes XRCC1 recruitment both at single-strand breaks globally across the genome and at sites of DNA replication stress. Finally, we show that the phosphate-binding pocket is required following DNA damage for XRCC1-dependent acceleration of DNA single-strand break repair, DNA base excision repair, and cell survival. These data support the hypothesis that poly (ADP-ribose) synthesis promotes XRCC1 recruitment at DNA damage sites and is important for XRCC1 function. PMID:26130715

  19. The XRCC1 phosphate-binding pocket binds poly (ADP-ribose) and is required for XRCC1 function

    Science.gov (United States)

    Breslin, Claire; Hornyak, Peter; Ridley, Andrew; Rulten, Stuart L.; Hanzlikova, Hana; Oliver, Antony W.; Caldecott, Keith W.

    2015-01-01

    Poly (ADP-ribose) is synthesized at DNA single-strand breaks and can promote the recruitment of the scaffold protein, XRCC1. However, the mechanism and importance of this process has been challenged. To address this issue, we have characterized the mechanism of poly (ADP-ribose) binding by XRCC1 and examined its importance for XRCC1 function. We show that the phosphate-binding pocket in the central BRCT1 domain of XRCC1 is required for selective binding to poly (ADP-ribose) at low levels of ADP-ribosylation, and promotes interaction with cellular PARP1. We also show that the phosphate-binding pocket is required for EGFP-XRCC1 accumulation at DNA damage induced by UVA laser, H2O2, and at sites of sub-nuclear PCNA foci, suggesting that poly (ADP-ribose) promotes XRCC1 recruitment both at single-strand breaks globally across the genome and at sites of DNA replication stress. Finally, we show that the phosphate-binding pocket is required following DNA damage for XRCC1-dependent acceleration of DNA single-strand break repair, DNA base excision repair, and cell survival. These data support the hypothesis that poly (ADP-ribose) synthesis promotes XRCC1 recruitment at DNA damage sites and is important for XRCC1 function. PMID:26130715

  20. Poly (ADP-ribose polymerase 1 is required for protein localization to Cajal body.

    Directory of Open Access Journals (Sweden)

    Elena Kotova

    2009-02-01

    Full Text Available Recently, the nuclear protein known as Poly (ADP-ribose Polymerase1 (PARP1 was shown to play a key role in regulating transcription of a number of genes and controlling the nuclear sub-organelle nucleolus. PARP1 enzyme is known to catalyze the transfer of ADP-ribose to a variety of nuclear proteins. At present, however, while we do know that the main acceptor for pADPr in vivo is PARP1 protein itself, by PARP1 automodification, the significance of PARP1 automodification for in vivo processes is not clear. Therefore, we investigated the roles of PARP1 auto ADP-ribosylation in dynamic nuclear processes during development. Specifically, we discovered that PARP1 automodification is required for shuttling key proteins into Cajal body (CB by protein non-covalent interaction with pADPr in vivo. We hypothesize that PARP1 protein shuttling follows a chain of events whereby, first, most unmodified PARP1 protein molecules bind to chromatin and accumulate in nucleoli, but then, second, upon automodification with poly(ADP-ribose, PARP1 interacts non-covalently with a number of nuclear proteins such that the resulting protein-pADPr complex dissociates from chromatin into CB.

  1. Ubiquitination independent of E1 and E2 enzymes by bacterial effectors.

    Science.gov (United States)

    Qiu, Jiazhang; Sheedlo, Michael J; Yu, Kaiwen; Tan, Yunhao; Nakayasu, Ernesto S; Das, Chittaranjan; Liu, Xiaoyun; Luo, Zhao-Qing

    2016-05-01

    Signalling by ubiquitination regulates virtually every cellular process in eukaryotes. Covalent attachment of ubiquitin to a substrate is catalysed by the E1, E2 and E3 three-enzyme cascade, which links the carboxy terminus of ubiquitin to the ε-amino group of, in most cases, a lysine of the substrate via an isopeptide bond. Given the essential roles of ubiquitination in the regulation of the immune system, it is not surprising that the ubiquitination network is a common target for diverse infectious agents. For example, many bacterial pathogens exploit ubiquitin signalling using virulence factors that function as E3 ligases, deubiquitinases or as enzymes that directly attack ubiquitin. The bacterial pathogen Legionella pneumophila utilizes approximately 300 effectors that modulate diverse host processes to create a permissive niche for its replication in phagocytes. Here we demonstrate that members of the SidE effector family of L. pneumophila ubiquitinate multiple Rab small GTPases associated with the endoplasmic reticulum. Moreover, we show that these proteins are capable of catalysing ubiquitination without the need for the E1 and E2 enzymes. A putative mono-ADP-ribosyltransferase motif critical for the ubiquitination activity is also essential for the role of the SidE family in intracellular bacterial replication in a protozoan host. The E1/E2-independent ubiquitination catalysed by these enzymes is energized by nicotinamide adenine dinucleotide, which activates ubiquitin by the formation of ADP-ribosylated ubiquitin. These results establish that ubiquitination can be catalysed by a single enzyme, the activity of which does not require ATP.

  2. Deficiency in Poly(ADP-ribose) Polymerase-1 (PARP-1) Accelerates Aging and Spontaneous Carcinogenesis in Mice

    Science.gov (United States)

    Piskunova, Tatiana S.; Yurova, Maria N.; Ovsyannikov, Anton I.; Semenchenko, Anna V.; Zabezhinski, Mark A.; Popovich, Irina G.; Wang, Zhao-Qi; Anisimov, Vladimir N.

    2008-01-01

    Genetic and biochemical studies have shown that PARP-1 and poly(ADP-ribosyl)ation play an important role in DNA repair, genomic stability, cell death, inflammation, telomere maintenance, and suppressing tumorigenesis, suggesting that the homeostasis of poly(ADP-ribosyl)ation and PARP-1 may also play an important role in aging. Here we show that PARP-1−/− mice exhibit a reduction of life span and a significant increase of population aging rate. Analysis of noninvasive parameters, including body weight gain, body temperature, estrous function, behavior, and a number of biochemical indices suggests the acceleration of biological aging in PARP-1−/− mice. The incidence of spontaneous tumors in both PARP-1−/− and PARP-1+/+ groups is similar; however, malignant tumors including uterine tumors, lung adenocarcinomas and hepatocellular carcinomas, develop at a significantly higher frequency in PARP-1−/− mice than PARP-1+/+ mice (72% and 49%, resp.; P < .05). In addition, spontaneous tumors appear earlier in PARP-1−/− mice compared to the wild type group. Histopathological studies revealed a wide spectrum of tumors in uterus, ovaries, liver, lungs, mammary gland, soft tissues, and lymphoid organs in both groups of the mice. These results demonstrate that inactivation of DNA repair gene PARP-1 in mice leads to acceleration of aging, shortened life span, and increased spontaneous carcinogenesis. PMID:19415146

  3. Deficiency in Poly(ADP-ribose Polymerase-1 (PARP-1 Accelerates Aging and Spontaneous Carcinogenesis in Mice

    Directory of Open Access Journals (Sweden)

    Tatiana S. Piskunova

    2008-01-01

    Full Text Available Genetic and biochemical studies have shown that PARP-1 and poly(ADP-ribosylation play an important role in DNA repair, genomic stability, cell death, inflammation, telomere maintenance, and suppressing tumorigenesis, suggesting that the homeostasis of poly(ADP-ribosylation and PARP-1 may also play an important role in aging. Here we show that PARP-1−/− mice exhibit a reduction of life span and a significant increase of population aging rate. Analysis of noninvasive parameters, including body weight gain, body temperature, estrous function, behavior, and a number of biochemical indices suggests the acceleration of biological aging in PARP-1−/− mice. The incidence of spontaneous tumors in both PARP-1−/− and PARP-1+/+ groups is similar; however, malignant tumors including uterine tumors, lung adenocarcinomas and hepatocellular carcinomas, develop at a significantly higher frequency in PARP-1−/− mice than PARP-1+/+ mice (72% and 49%, resp.; < .05. In addition, spontaneous tumors appear earlier in PARP-1−/− mice compared to the wild type group. Histopathological studies revealed a wide spectrum of tumors in uterus, ovaries, liver, lungs, mammary gland, soft tissues, and lymphoid organs in both groups of the mice. These results demonstrate that inactivation of DNA repair gene PARP-1 in mice leads to acceleration of aging, shortened life span, and increased spontaneous carcinogenesis.

  4. Antibody-Directed Phototherapy (ADP

    Directory of Open Access Journals (Sweden)

    M. Adil Butt

    2013-04-01

    Full Text Available Photodynamic therapy (PDT is a clinically-approved but rather under-exploited treatment modality for cancer and pre-cancerous superficial lesions. It utilises a cold laser or LED to activate a photochemical reaction between a light activated drug (photosensitiser-drug and oxygen to generate cytotoxic oxygen species. These free radical species damage cellular components leading to cell death. Despite its benefits, the complexity, limited potency and side effects of PDT have led to poor general usage. However, the research area is very active with an increasing understanding of PDT-related cell biology, photophysics and significant progress in molecular targeting of disease. Monoclonal antibody therapy is maturing and the next wave of antibody therapies includes antibody-drug conjugates (ADCs, which promise to be more potent and curable. These developments could lift antibody-directed phototherapy (ADP to success. ADP promises to increase specificity and potency and improve drug pharmacokinetics, thus delivering better PDT drugs whilst retaining its other benefits. Whole antibody conjugates with first generation ADP-drugs displayed problems with aggregation, poor pharmacokinetics and loss of immuno-reactivity. However, these early ADP-drugs still showed improved selectivity and potency. Improved PS-drug chemistry and a variety of conjugation strategies have led to improved ADP-drugs with retained antibody and PS-drug function. More recently, recombinant antibody fragments have been used to deliver ADP-drugs with superior drug loading, more favourable pharmacokinetics, enhanced potency and target cell selectivity. These improvements offer a promise of better quality PDT drugs.

  5. Identification of class II ADP-ribosylation factors as cellular factors required for hepatitis C virus replication.

    Science.gov (United States)

    Farhat, Rayan; Séron, Karin; Ferlin, Juliette; Fénéant, Lucie; Belouzard, Sandrine; Goueslain, Lucie; Jackson, Catherine L; Dubuisson, Jean; Rouillé, Yves

    2016-08-01

    GBF1 is a host factor required for hepatitis C virus (HCV) replication. GBF1 functions as a guanine nucleotide exchange factor for G-proteins of the Arf family, which regulate membrane dynamics in the early secretory pathway and the metabolism of cytoplasmic lipid droplets. Here we established that the Arf-guanine nucleotide exchange factor activity of GBF1 is critical for its function in HCV replication, indicating that it promotes viral replication by activating one or more Arf family members. Arf involvement was confirmed with the use of two dominant negative Arf1 mutants. However, siRNA-mediated depletion of Arf1, Arf3 (class I Arfs), Arf4 or Arf5 (class II Arfs), which potentially interact with GBF1, did not significantly inhibit HCV infection. In contrast, the simultaneous depletion of both Arf4 and Arf5, but not of any other Arf pair, imposed a significant inhibition of HCV infection. Interestingly, the simultaneous depletion of both Arf4 and Arf5 had no impact on the activity of the secretory pathway and induced a compaction of the Golgi and an accumulation of lipid droplets. A similar phenotype of lipid droplet accumulation was also observed when GBF1 was inhibited by brefeldin A. In contrast, the simultaneous depletion of both Arf1 and Arf4 resulted in secretion inhibition and Golgi scattering, two actions reminiscent of GBF1 inhibition. We conclude that GBF1 could regulate different metabolic pathways through the activation of different pairs of Arf proteins. PMID:26814617

  6. Differential and Concordant Roles for Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribose) in Regulating WRN and RECQL5 Activities.

    Science.gov (United States)

    Khadka, Prabhat; Hsu, Joseph K; Veith, Sebastian; Tadokoro, Takashi; Shamanna, Raghavendra A; Mangerich, Aswin; Croteau, Deborah L; Bohr, Vilhelm A

    2015-12-01

    Poly(ADP-ribose) (PAR) polymerase 1 (PARP1) catalyzes the poly(ADP-ribosyl)ation (PARylation) of proteins, a posttranslational modification which forms the nucleic acid-like polymer PAR. PARP1 and PAR are integral players in the early DNA damage response, since PARylation orchestrates the recruitment of repair proteins to sites of damage. Human RecQ helicases are DNA unwinding proteins that are critical responders to DNA damage, but how their recruitment and activities are regulated by PARPs and PAR is poorly understood. Here we report that all human RecQ helicases interact with PAR noncovalently. Furthermore, we define the effects that PARP1, PARylated PARP1, and PAR have on RECQL5 and WRN, using both in vitro and in vivo assays. We show that PARylation is involved in the recruitment of RECQL5 and WRN to laser-induced DNA damage and that RECQL5 and WRN have differential responses to PARylated PARP1 and PAR. Furthermore, we show that the loss of RECQL5 or WRN resulted in increased sensitivity to PARP inhibition. In conclusion, our results demonstrate that PARP1 and PAR actively, and in some instances differentially, regulate the activities and cellular localization of RECQL5 and WRN, suggesting that PARylation acts as a fine-tuning mechanism to coordinate their functions in time and space during the genotoxic stress response. PMID:26391948

  7. Differential and Concordant Roles for Poly(ADP-Ribose) Polymerase 1 and Poly(ADP-Ribose) in Regulating WRN and RECQL5 Activities.

    Science.gov (United States)

    Khadka, Prabhat; Hsu, Joseph K; Veith, Sebastian; Tadokoro, Takashi; Shamanna, Raghavendra A; Mangerich, Aswin; Croteau, Deborah L; Bohr, Vilhelm A

    2015-12-01

    Poly(ADP-ribose) (PAR) polymerase 1 (PARP1) catalyzes the poly(ADP-ribosyl)ation (PARylation) of proteins, a posttranslational modification which forms the nucleic acid-like polymer PAR. PARP1 and PAR are integral players in the early DNA damage response, since PARylation orchestrates the recruitment of repair proteins to sites of damage. Human RecQ helicases are DNA unwinding proteins that are critical responders to DNA damage, but how their recruitment and activities are regulated by PARPs and PAR is poorly understood. Here we report that all human RecQ helicases interact with PAR noncovalently. Furthermore, we define the effects that PARP1, PARylated PARP1, and PAR have on RECQL5 and WRN, using both in vitro and in vivo assays. We show that PARylation is involved in the recruitment of RECQL5 and WRN to laser-induced DNA damage and that RECQL5 and WRN have differential responses to PARylated PARP1 and PAR. Furthermore, we show that the loss of RECQL5 or WRN resulted in increased sensitivity to PARP inhibition. In conclusion, our results demonstrate that PARP1 and PAR actively, and in some instances differentially, regulate the activities and cellular localization of RECQL5 and WRN, suggesting that PARylation acts as a fine-tuning mechanism to coordinate their functions in time and space during the genotoxic stress response.

  8. Enhanced production and action of cyclic ADP-ribose during oxidative stress in small bovine coronary arterial smooth muscle.

    Science.gov (United States)

    Zhang, Andrew Y; Yi, Fan; Teggatz, Eric G; Zou, Ai-Ping; Li, Pin-Lan

    2004-03-01

    Recent studies in our lab and by others have indicated that cyclic ADP-ribose (cADPR) as a novel second messenger is importantly involved in vasomotor response in various vascular beds. However, the mechanism regulating cADPR production and actions remains poorly understood. The present study determined whether changes in redox status influence the production and action of cADPR in coronary arterial smooth muscle cells (CASMCs) and thereby alters vascular tone in these arteries. HPLC analyses demonstrated that xanthine (X, 40 microM)/xanthine oxidase (XO, 0.1 U/ml), a superoxide-generating system, increased the ADP-ribosyl cyclase activity by 59% in freshly isolated bovine CASMCs. However, hydrogen peroxide (H2O2, 1-100 microM) had no significant effect on ADP-ribosyl cyclase activity. In these CASMCs, X/XO produced a rapid increase in [Ca2+]i (Delta[Ca2+]i=201 nM), which was significantly attenuated by a cADPR antagonist, 8-Br-cADPR. Both inhibition of cADPR production by nicotinamide (Nicot) and blockade of Ca2+-induced Ca2+ release (CICR) by tetracaine (TC) and ryanodine (Rya) significantly reduced X/XO-induced rapid Ca2+ responses. In isolated, perfused, and pressurized small bovine coronary arteries, X at 2.5-80 microM with a fixed XO level produced a concentration-dependent vasoconstriction with a maximal decrease in arterial diameter of 45%. This X/XO-induced vasoconstriction was significantly attenuated by 8-Br-cADPR, Nicot, TC, or Rya. We conclude that superoxide activates cADPR production, and thereby mobilizes intracellular Ca2+ from the SR and produces vasoconstriction in coronary arteries.

  9. Sumoylation of poly(ADP-ribose) polymerase 1 inhibits its acetylation and restrains transcriptional coactivator function.

    Science.gov (United States)

    Messner, Simon; Schuermann, David; Altmeyer, Matthias; Kassner, Ingrid; Schmidt, Darja; Schär, Primo; Müller, Stefan; Hottiger, Michael O

    2009-11-01

    Poly(ADP-ribose) polymerase 1 (PARP1) is a chromatin-associated nuclear protein and functions as a molecular stress sensor. At the cellular level, PARP1 has been implicated in a wide range of processes, such as maintenance of genome stability, cell death, and transcription. PARP1 functions as a transcriptional coactivator of nuclear factor kappaB (NF-kappaB) and hypoxia inducible factor 1 (HIF1). In proteomic studies, PARP1 was found to be modified by small ubiquitin-like modifiers (SUMOs). Here, we characterize PARP1 as a substrate for modification by SUMO1 and SUMO3, both in vitro and in vivo. PARP1 is sumoylated at the single lysine residue K486 within its automodification domain. Interestingly, modification of PARP1 with SUMO does not affect its ADP-ribosylation activity but completely abrogates p300-mediated acetylation of PARP1, revealing an intriguing crosstalk of sumoylation and acetylation on PARP1. Genetic complementation of PARP1-depleted cells with wild-type and sumoylation-deficient PARP1 revealed that SUMO modification of PARP1 restrains its transcriptional coactivator function and subsequently reduces gene expression of distinct PARP1-regulated target genes.

  10. Crystallographic Analysis of Tapering of ADP Crystallites

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    On the basis of crystallographic characteristics of ADP (ammonium dihydrogen phosphate) crystals and the selected growth conditions, the growth habit of ADP crystals was studied. In comparison with pyramidal planes, the growth rate of prismatic faces is slower and more sensitive to the additives and impurities for ADP crystals. When the supersaturation is low, the advance of growth steps on prismatic face can be blocked by ethanol or impurities, the crystal morphology is changed from the tetragonal prism to shuttle (i.e., the tapered shape). The tapering formation of ADP crystallites was structurally studied in a novel view.

  11. Study of the Five Rickettsia prowazekii Proteins Annotated as ATP/ADP Translocases (Tlc): Only Tlc1 Transports ATP/ADP, While Tlc4 and Tlc5 Transport Other Ribonucleotides

    OpenAIRE

    Audia, Jonathon P.; Winkler, Herbert H.

    2006-01-01

    The obligate intracytoplasmic pathogen Rickettsia prowazekii relies on the transport of many essential compounds from the cytoplasm of the eukaryotic host cell in lieu of de novo synthesis, an evolutionary outcome undoubtedly linked to obligatory growth in this metabolite-replete niche. The paradigm for the study of rickettsial transport systems is the ATP/ADP translocase Tlc1, which exchanges bacterial ADP for host cell ATP as a source of energy, rather than as a source of adenylate. Interes...

  12. Rho-modifying bacterial protein toxins from Photorhabdus species.

    Science.gov (United States)

    Jank, Thomas; Lang, Alexander E; Aktories, Klaus

    2016-06-15

    Photorhabdus bacteria live in symbiosis with entomopathogenic nematodes. The nematodes invade insect larvae, where they release the bacteria, which then produce toxins to kill the insects. Recently, the molecular mechanisms of some toxins from Photorhabdus luminescens and asymbiotica have been elucidated, showing that GTP-binding proteins of the Rho family are targets. The tripartite Tc toxin PTC5 from P. luminescens activates Rho proteins by ADP-ribosylation of a glutamine residue, which is involved in GTP hydrolysis, while PaTox from Photorhabdus asymbiotica inhibits the activity of GTPases by N-acetyl-glucosaminylation at tyrosine residues and activates Rho proteins indirectly by deamidation of heterotrimeric G proteins.

  13. Adenosine Diphosphate Ribosylation Factor-GTPaseActivating Protein Stimulates the Transport of AUX1Endosome, Which Relies on Actin Cytoskeletal Organization in Rice Root DevelopmentF

    Institute of Scientific and Technical Information of China (English)

    Cheng Du; Yunyuan XU; Yingdian Wang; Kang Chong

    2011-01-01

    Polar auxin transport,which depends on polarized subcellular distribution of AUXIN RESISTANT 1/LIKE AUX1 (AUX1/LAX) influx carriers and PIN-FORMED (PIN) efflux carriers,mediates various processes of plant growth and development.Endosomal recycling of PIN1 is mediated by an adenosine diphosphate (ADP)ribosylation factor (ARF)-GTPase exchange factor protein,GNOM.However,the mediation of auxin influx carrier recycling is poorly understood.Here,we report that overexpression of OsAGAP,an ARF-GTPase-activating protein in rice,stimulates vesicle transport from the plasma membrane to the Golgi apparatus in protoplasts and transgenic plants and induces the accumulation of early endosomes and AUX1.AUX1 endosomes could partially colocalize with FM4-64 labeled early endosome after actin disruption.Furthermore,OsAGAP is involved in actin cytoskeletal organization,and its overexpression tends to reduce the thickness and bundling of actin filaments.Fluorescence recovery after photobleaching analysis revealed exocytosis of the AUX1 recycling endosome was not affected in the OsAGAP overexpression cells,and was only slightly promoted when the actin filaments were completely disrupted by Lat B.Thus,we propose that AUX1 accumulation in the OsAGAP overexpression and actin disrupted cells may be due to the fact that endocytosis of the auxin influx carrier AUX1 early endosome was greatly promoted by actin cytoskeleton disruption.

  14. Poly(ADP-ribose polymerase 1 is indispensable for transforming growth factor-β Induced Smad3 activation in vascular smooth muscle cell.

    Directory of Open Access Journals (Sweden)

    Dan Huang

    Full Text Available BACKGROUND: Transforming growth factor type-β (TGF-β/Smad pathway plays an essential role in vascular fibrosis. Reactive oxygen species (ROS generation also mediates TGF-β signaling-induced vascular fibrosis, suggesting that some sort of interaction exists between Smad and redox pathways. However, the underlying molecular mechanism is largely unknown. This study aims to investigate the influence of poly(ADP-ribose polymerase 1 (PARP1, a downstream effector of ROS, on TGF-β signaling transduction through Smad3 pathway in rat vascular smooth muscle cells (VSMCs. METHODS AND RESULTS: TGF-β1 treatment promoted PARP1 activation through induction of ROS generation in rat VSMCs. TGF-β1-induced phosphorylation and nuclear accumulation of Smad3 was prevented by treatment of cells with PARP inhibitor, 3-aminobenzamide (3AB or N-(6-oxo-5,6-dihydrophenanthridin-2-yl-2-(N,N-dimethylaminoacetami (PJ34, or PARP1 siRNA. TGF-β1 treatment promoted poly(ADP-ribosylation of Smad3 via activation of PARP1 in the nucleus. Poly(ADP-ribosylation enhanced Smad-Smad binding element (SBE complex formation in nuclear extracts and increased DNA binding activity of Smad3. Pretreatment with 3AB, PJ34, or PARP1 siRNA prevented TGF-β1-induced Smad3 transactivation and expression of Smad3 target genes, including collagen Iα1, collagen IIIα1 and tissue inhibitor of metalloproteinase 1, in rat VSMCs. CONCLUSIONS: PARP1 is indispensable for TGF-β1 induced Smad3 activation in rat VSMCs. Targeting PARP1 may be a promising therapeutic approach against vascular diseases induced by dysregulation of TGF-β/Smad3 pathway.

  15. Transcriptional regulation by Poly(ADP-ribose polymerase-1 during T cell activation

    Directory of Open Access Journals (Sweden)

    Parrilla Pascual

    2008-04-01

    Full Text Available Abstract Background Accumulating evidence suggests an important role for the enzyme poly(ADP-ribose polymerase-1 (PARP-1 as an integral part of the gene expression regulatory machinery during development and in response to specific cellular signals. PARP-1 might modulate gene expression through its catalytic activity leading to poly(ADP-ribosylation of nuclear proteins or by its physical association with relevant proteins. Recently, we have shown that PARP-1 is activated during T cell activation. However, the proposed role of PARP-1 in reprogramming T cell gene expression upon activation remains largely unexplored. Results In the present study we use oligonucleotide microarray analysis to gain more insight into the role played by PARP-1 during the gene expression reprogramming that takes place in T cells upon activation with anti-CD3 stimulation alone, or in combination with anti-CD28 co-stimulation. We have identified several groups of genes with expression modulated by PARP-1. The expression of 129 early-response genes to anti-CD3 seems to be regulated by PARP-1 either in a positive (45 genes or in a negative manner (84 genes. Likewise, in the presence of co-stimulation (anti-CD3 + anti-CD28 stimulation, the expression of 203 genes is also regulated by PARP-1 either up (173 genes or down (30 genes. Interestingly, PARP-1 deficiency significantly alters expression of genes associated with the immune response such as chemokines and genes involved in the Th1/Th2 balance. Conclusion This study provides new insights into changes in gene expression mediated by PARP-1 upon T cell activation. Pathway analysis of PARP-1 as a nuclear signalling molecule in T cells would be of relevance for the future development of new therapeutic approaches targeting PARP-1 in the acquired immune response.

  16. In silico characterization of the family of PARP-like poly(ADP-ribosyltransferases (pARTs

    Directory of Open Access Journals (Sweden)

    Dittmar Katharina

    2005-10-01

    Full Text Available Abstract Background ADP-ribosylation is an enzyme-catalyzed posttranslational protein modification in which mono(ADP-ribosyltransferases (mARTs and poly(ADP-ribosyltransferases (pARTs transfer the ADP-ribose moiety from NAD onto specific amino acid side chains and/or ADP-ribose units on target proteins. Results Using a combination of database search tools we identified the genes encoding recognizable pART domains in the public genome databases. In humans, the pART family encompasses 17 members. For 16 of these genes, an orthologue exists also in the mouse, rat, and pufferfish. Based on the degree of amino acid sequence similarity in the catalytic domain, conserved intron positions, and fused protein domains, pARTs can be divided into five major subgroups. All six members of groups 1 and 2 contain the H-Y-E trias of amino acid residues found also in the active sites of Diphtheria toxin and Pseudomonas exotoxin A, while the eleven members of groups 3 – 5 carry variations of this motif. The pART catalytic domain is found associated in Lego-like fashion with a variety of domains, including nucleic acid-binding, protein-protein interaction, and ubiquitylation domains. Some of these domain associations appear to be very ancient since they are observed also in insects, fungi, amoebae, and plants. The recently completed genome of the pufferfish T. nigroviridis contains recognizable orthologues for all pARTs except for pART7. The nearly completed albeit still fragmentary chicken genome contains recognizable orthologues for twelve pARTs. Simpler eucaryotes generally contain fewer pARTs: two in the fly D. melanogaster, three each in the mosquito A. gambiae, the nematode C. elegans, and the ascomycete microfungus G. zeae, six in the amoeba E. histolytica, nine in the slime mold D. discoideum, and ten in the cress plant A. thaliana. GenBank contains two pART homologues from the large double stranded DNA viruses Chilo iridescent virus and Bacteriophage Aeh1

  17. 45 CFR 95.619 - Use of ADP systems.

    Science.gov (United States)

    2010-10-01

    ... Data Processing Equipment and Services-Conditions for Federal Financial Participation (FFP) Specific Conditions for Ffp § 95.619 Use of ADP systems. ADP systems designed, developed, or installed with FFP...

  18. Dielectric, thermal and mechanical properties of ADP doped PVA composites

    Science.gov (United States)

    Naik, Jagadish; Bhajantri, R. F.; Ravindrachary, V.; Rathod, Sunil G.; Sheela, T.; Naik, Ishwar

    2015-06-01

    Polymer composites of poly(vinyl alcohol) (PVA), doped with different concentrations of ammonium dihydrogen phosphate (ADP) has been prepared by solution casting. The formation of complexation between ADP and PVA was confirmed with the help of Fourier transforms infrared (FTIR) spectroscopy. Thermogravimetric analysis (TGA) shows thermal stability of the prepared composites. Impedance analyzer study revealed the increase in dielectric constant and loss with increase the ADP concentration and the strain rate of the prepared composites decreases with ADP concentration.

  19. 42 CFR 457.230 - FFP for State ADP expenditures.

    Science.gov (United States)

    2010-10-01

    ... procedures regarding the availability of FFP for ADP expenditures are in 45 CFR part 74, 45 CFR part 95... 42 Public Health 4 2010-10-01 2010-10-01 false FFP for State ADP expenditures. 457.230 Section 457...; Reduction of Federal Medical Payments § 457.230 FFP for State ADP expenditures. FFP is available for...

  20. Poly(ADP-ribosyl)ation links the chromatin remodeler SMARCA5/SNF2H to RNF168-dependent DNA damage signaling

    NARCIS (Netherlands)

    G. Smeenk (Godelieve); W.W. Wiegant (Wouter); J.A. Marteijn (Jurgen); M.S. Luijsterburg (Martijn); N. Sroczynski (Nicholas); T. Costelloe (Thomas); R. Romeijn (Ron); A. Pastink (Albert); N. Mailand (Niels); W. Vermeulen (Wim); H. van Attikum (Haico)

    2013-01-01

    textabstractIonizing radiation (IR)-induced DNA double-strand breaks (DSBs) arising in native chromatin elicit an RNF8/RNF168-dependent ubiquitylation response, which triggers the recruitment of various repair factors. Precisely how this response is regulated in the context of chromatin remains larg

  1. ADP Analysis project for the Human Resources Management Division

    Science.gov (United States)

    Tureman, Robert L., Jr.

    1993-01-01

    The ADP (Automated Data Processing) Analysis Project was conducted for the Human Resources Management Division (HRMD) of NASA's Langley Research Center. The three major areas of work in the project were computer support, automated inventory analysis, and an ADP study for the Division. The goal of the computer support work was to determine automation needs of Division personnel and help them solve computing problems. The goal of automated inventory analysis was to find a way to analyze installed software and usage on a Macintosh. Finally, the ADP functional systems study for the Division was designed to assess future HRMD needs concerning ADP organization and activities.

  2. Interaction of ADP, atractyloside, and gummiferin on the ADP translocase of the inner mitochondrial membrane

    Energy Technology Data Exchange (ETDEWEB)

    Vignais, P.V.; Vignais, P.M.; Defaye, G.; Lauquin, G.; Doussiere, J.; Chabert, J.; Brandolin, G.

    1972-05-01

    From international conference on mechanism in bioenergetica; Bari, Italy (1 May 1972). Two specific inhibitors of the adenine nucleotide translocation, gummiferin (GUM), identified to 4-carboxyatractyloside and atractyloside (ATR), were labeled with /sup 35/S and their binding properties to whole mitochondria and inner mitochondrial membrane vesicles used to monitor changes of membrane conformation induced by ADP. (auth)

  3. Study of the five Rickettsia prowazekii proteins annotated as ATP/ADP translocases (Tlc): Only Tlc1 transports ATP/ADP, while Tlc4 and Tlc5 transport other ribonucleotides.

    Science.gov (United States)

    Audia, Jonathon P; Winkler, Herbert H

    2006-09-01

    The obligate intracytoplasmic pathogen Rickettsia prowazekii relies on the transport of many essential compounds from the cytoplasm of the eukaryotic host cell in lieu of de novo synthesis, an evolutionary outcome undoubtedly linked to obligatory growth in this metabolite-replete niche. The paradigm for the study of rickettsial transport systems is the ATP/ADP translocase Tlc1, which exchanges bacterial ADP for host cell ATP as a source of energy, rather than as a source of adenylate. Interestingly, the R. prowazekii genome encodes four open reading frames that are highly homologous to the well-characterized ATP/ADP translocase Tlc1. Therefore, by annotation, the R. prowazekii genome encodes a total of five ATP/ADP translocases: Tlc1, Tlc2, Tlc3, Tlc4, and Tlc5. We have confirmed by quantitative reverse transcriptase PCR that mRNAs corresponding to all five tlc homologues are expressed in R. prowazekii growing in L-929 cells and have shown their heterologous protein expression in Escherichia coli, suggesting that none of the tlc genes are pseudogenes in the process of evolutionary meltdown. However, we demonstrate by heterologous expression in E. coli that only Tlc1 functions as an ATP/ADP transporter. A survey of nucleotides and nucleosides has determined that Tlc4 transports CTP, UTP, and GDP. Intriguingly, although GTP was not transported by Tlc4, it was an inhibitor of CTP and UTP uptake and demonstrated a K(i) similar to that of GDP. In addition, we demonstrate that Tlc5 transports GTP and GDP. We postulate that Tlc4 and Tlc5 serve the primary function of maintaining intracellular pools of nucleotides for rickettsial nucleic acid biosynthesis and do not provide the cell with nucleoside triphosphates as an energy source, as is the case for Tlc1. Although heterologous expression of Tlc2 and Tlc3 was observed in E. coli, we were unable to identify substrates for these proteins. PMID:16923893

  4. Issues on stability of ADP feedback controllers for dynamical systems.

    Science.gov (United States)

    Balakrishnan, S N; Ding, Jie; Lewis, Frank L

    2008-08-01

    This paper traces the development of neural-network (NN)-based feedback controllers that are derived from the principle of adaptive/approximate dynamic programming (ADP) and discusses their closed-loop stability. Different versions of NN structures in the literature, which embed mathematical mappings related to solutions of the ADP-formulated problems called "adaptive critics" or "action-critic" networks, are discussed. Distinction between the two classes of ADP applications is pointed out. Furthermore, papers in "model-free" development and model-based neurocontrollers are reviewed in terms of their contributions to stability issues. Recent literature suggests that work in ADP-based feedback controllers with assured stability is growing in diverse forms. PMID:18632377

  5. ADP Security Plan: Rice Lake National Wildlife Refuge

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This ADP Security Plan established the policies and procedures to protect the integrity of computer-based resources at this site. Specifically, the purpose of this...

  6. Latonduine Analogs Restore F508del-Cystic Fibrosis Transmembrane Conductance Regulator Trafficking through the Modulation of Poly-ADP Ribose Polymerase 3 and Poly-ADP Ribose Polymerase 16 Activity.

    Science.gov (United States)

    Carlile, Graeme W; Robert, Renaud; Matthes, Elizabeth; Yang, Qi; Solari, Roberto; Hatley, Richard; Edge, Colin M; Hanrahan, John W; Andersen, Raymond; Thomas, David Y; Birault, Véronique

    2016-08-01

    Cystic fibrosis (CF) is a major lethal genetic disease caused by mutations in the CF transmembrane conductance regulator gene (CFTR). This encodes a chloride ion channel on the apical surface of epithelial cells. The most common mutation in CFTR (F508del-CFTR) generates a protein that is misfolded and retained in the endoplasmic reticulum. Identifying small molecules that correct this CFTR trafficking defect is a promising approach in CF therapy. However, to date only modest efficacy has been reported for correctors in clinical trials. We identified the marine sponge metabolite latonduine as a corrector. We have now developed a series of latonduine derivatives that are more potent F508del-CFTR correctors with one (MCG315 [2,3-dihydro-1H-2-benzazepin-1-one]) having 10-fold increased corrector activity and an EC50 of 72.25 nM. We show that the latonduine analogs inhibit poly-ADP ribose polymerase (PARP) isozymes 1, 3, and 16. Further our molecular modeling studies point to the latonduine analogs binding to the PARP nicotinamide-binding domain. We established the relationship between the ability of the latonduine analogs to inhibit PARP-16 and their ability to correct F508del-CFTR trafficking. We show that latonduine can inhibit both PARP-3 and -16 and that this is necessary for CFTR correction. We demonstrate that latonduine triggers correction by regulating the activity of the unfolded protein response activator inositol-requiring enzyme (IRE-1) via modulation of the level of its ribosylation by PARP-16. These results establish latonduines novel site of action as well as its proteostatic mechanism of action. PMID:27193581

  7. Regulation of the RhoA/ROCK/AKT/β-catenin pathway by arginine-specific ADP-ribosytransferases 1 promotes migration and epithelial-mesenchymal transition in colon carcinoma.

    Science.gov (United States)

    Song, Guang-Lin; Jin, Cong-Cong; Zhao, Wei; Tang, Yi; Wang, Ya-Lan; Li, Ming; Xiao, Ming; Li, Xian; Li, Qing-Shu; Lin, Xiao; Chen, Wen-Wen; Kuang, Jing

    2016-08-01

    Arginine-specific ADP-ribosytransferases 1 (ART1) is able to modify the arginine of specific proteins by mono-ADP-ribosylation. We previously reported that the expression of ART1 in human colon adenocarcinoma tissues was higher than in adjacent tissues. Herein, we primarily revealed that ART1 could regulate the epithelial-mesenchymal transition (EMT) and, therefore, the development of colon carcinoma. In CT26 cells, which overexpressed ART1 by lentiviral transfection, the following were promoted: alterations of spindle-like non-polarization, expression of EMT inducers and mesenchymal markers, migration, invasion and adhesion. However, epithelial marker expression was decreased. Correspondingly, knockdown of ART1 in CT26 cells had the opposite effects. The effect of ART1 on EMT and carcinoma metastasis was also verified in a liver metastasis model of BALB/c mice. To further explore the molecular mechanism of ART1 in EMT, CT26 cells were treated with several specific inhibitors and gene silencing. Our data suggest that ART1 could regulate EMT by regulating the RhoA/ROCK1/AKT/β-catenin pathway and its downstream factors (snail1, vimentin, N-cadherin and E-cadherin) and that it therefore plays an important role in the progression of colon carcinoma. PMID:27277835

  8. Functional characterisation of an Arabidopsis gene strongly induced by ionising radiation: the gene coding the poly(ADP-ribose)polymerase-1 (AthPARP-1)

    International Nuclear Information System (INIS)

    Arabidopsis thaliana, the model-system in plant genetics, has been used to study the responses to DNA damage, experimentally introduced by γ-irradiation. We have characterised a radiation-induced gene coding a 111 kDa protein, AthPARP-1, homologous to the human poly(ADP-ribose)polymerase-1 (hPARP-1). As hPARP-1 is composed by three functional domain with characteristic motifs, AthPARP-1 binds to DNA bearing single-strand breaks and shows DNA damage-dependent poly(ADP-ribosyl)ation. The preferential expression of AthPARP-1 in mitotically active tissues is in agreement with a potential role in the maintenance of genome integrity during DNA replication, as proposed for its human counterpart. Transcriptional gene activation by ionising radiation of AthPARP-1 and AthPARP-2 genes is to date plant specific activation. Our expression analyses after exposure to various stress indicate that 1) AthPARP-1 and AthPARP-2 play an important role in the response to DNA lesions, particularly they are activated by genotoxic agents implicating the BER DNA repair pathway 2) AthPARP-2 gene seems to play an additional role in the signal transduction induced by oxidative stress 3) the observed expression profile of AthPARP-1 is in favour of the regulation of AthPARP-1 gene expression at the level of transcription and translation. This mode of regulation of AthPARP-1 protein biosynthesis, clearly distinct from that observed in animals, needs the implication of a so far unidentified transcription factor that is activated by the presence of DNA lesions. The major outcome of this work resides in the isolation and characterisation of such new transcription factor, which will provide new insight on the regulation of plant gene expression by genotoxic stress. (author)

  9. Cholera- and anthrax-like toxins are among several new ADP-ribosyltransferases.

    Directory of Open Access Journals (Sweden)

    Robert J Fieldhouse

    Full Text Available Chelt, a cholera-like toxin from Vibrio cholerae, and Certhrax, an anthrax-like toxin from Bacillus cereus, are among six new bacterial protein toxins we identified and characterized using in silico and cell-based techniques. We also uncovered medically relevant toxins from Mycobacterium avium and Enterococcus faecalis. We found agriculturally relevant toxins in Photorhabdus luminescens and Vibrio splendidus. These toxins belong to the ADP-ribosyltransferase family that has conserved structure despite low sequence identity. Therefore, our search for new toxins combined fold recognition with rules for filtering sequences--including a primary sequence pattern--to reduce reliance on sequence identity and identify toxins using structure. We used computers to build models and analyzed each new toxin to understand features including: structure, secretion, cell entry, activation, NAD+ substrate binding, intracellular target binding and the reaction mechanism. We confirmed activity using a yeast growth test. In this era where an expanding protein structure library complements abundant protein sequence data--and we need high-throughput validation--our approach provides insight into the newest toxin ADP-ribosyltransferases.

  10. ADP1 affects plant architecture by regulating local auxin biosynthesis.

    Science.gov (United States)

    Li, Ruixi; Li, Jieru; Li, Shibai; Qin, Genji; Novák, Ondřej; Pěnčík, Aleš; Ljung, Karin; Aoyama, Takashi; Liu, Jingjing; Murphy, Angus; Gu, Hongya; Tsuge, Tomohiko; Qu, Li-Jia

    2014-01-01

    Plant architecture is one of the key factors that affect plant survival and productivity. Plant body structure is established through the iterative initiation and outgrowth of lateral organs, which are derived from the shoot apical meristem and root apical meristem, after embryogenesis. Here we report that ADP1, a putative MATE (multidrug and toxic compound extrusion) transporter, plays an essential role in regulating lateral organ outgrowth, and thus in maintaining normal architecture of Arabidopsis. Elevated expression levels of ADP1 resulted in accelerated plant growth rate, and increased the numbers of axillary branches and flowers. Our molecular and genetic evidence demonstrated that the phenotypes of plants over-expressing ADP1 were caused by reduction of local auxin levels in the meristematic regions. We further discovered that this reduction was probably due to decreased levels of auxin biosynthesis in the local meristematic regions based on the measured reduction in IAA levels and the gene expression data. Simultaneous inactivation of ADP1 and its three closest homologs led to growth retardation, relative reduction of lateral organ number and slightly elevated auxin level. Our results indicated that ADP1-mediated regulation of the local auxin level in meristematic regions is an essential determinant for plant architecture maintenance by restraining the outgrowth of lateral organs. PMID:24391508

  11. ADP1 affects plant architecture by regulating local auxin biosynthesis.

    Directory of Open Access Journals (Sweden)

    Ruixi Li

    2014-01-01

    Full Text Available Plant architecture is one of the key factors that affect plant survival and productivity. Plant body structure is established through the iterative initiation and outgrowth of lateral organs, which are derived from the shoot apical meristem and root apical meristem, after embryogenesis. Here we report that ADP1, a putative MATE (multidrug and toxic compound extrusion transporter, plays an essential role in regulating lateral organ outgrowth, and thus in maintaining normal architecture of Arabidopsis. Elevated expression levels of ADP1 resulted in accelerated plant growth rate, and increased the numbers of axillary branches and flowers. Our molecular and genetic evidence demonstrated that the phenotypes of plants over-expressing ADP1 were caused by reduction of local auxin levels in the meristematic regions. We further discovered that this reduction was probably due to decreased levels of auxin biosynthesis in the local meristematic regions based on the measured reduction in IAA levels and the gene expression data. Simultaneous inactivation of ADP1 and its three closest homologs led to growth retardation, relative reduction of lateral organ number and slightly elevated auxin level. Our results indicated that ADP1-mediated regulation of the local auxin level in meristematic regions is an essential determinant for plant architecture maintenance by restraining the outgrowth of lateral organs.

  12. Plant extracts inhibit ADP-induced platelet activation in humans: their potential therapeutic role as ADP antagonists.

    Science.gov (United States)

    Jagroop, Indera Anita

    2014-01-01

    Adenosine diphosphate (ADP) plays a pivotal role in platelet activation. Platelet hyperactivity is associated with vascular disease and also has a key role in haemostasis and thrombosis. ADP activates platelets through three purinoceptor subtypes, the G(q)-coupled P2Y(1) receptor, G(i)-coupled P2Y(12) receptor and P2X(1) ligand-gated cation channel. Platelet ADP purinergic receptors are therefore suitable targets for antiplatelet drugs. Thienopyridines such as clopidogrel and ticlopidine, as well as other ADP receptor antagonists like prasugrel, ticagrelor, cangrelor and elinogrel have demonstrated clinical benefits via the inhibition of the selective purinergic ADP receptor, P2Y(12). However, they still have limitations in their mode of action and efficacy, like increased risk of bleeding. Thus, the ongoing pursuit to develop newer and more effective antiplatelet agents continues. There is a growing interest in the purinergic antiplatelet properties exhibited by plant extracts. This article considers the following: pomolic acid isolated from Licania pittieri, brazilin isolated from the heartwood of Caesalpinia sappan L, phylligenin isolated from the twigs of Muraltia vulpina, bark oil of Gonystylus velutinus, seed and bark extracts from Aesculus hippocastanum L. and red wine phenolics and catechins isolated from green tea. Moreover, the method used to investigate platelet purinergic receptors should be considered, since using a more sensitive, high-resolution platelet sizer can sometimes detect platelet variations when the light transmission method was not able to do so. The exact mechanisms by which these plant extracts work need further investigation. They all however inhibit ADP-induced activation in human platelets. This could explain, at least in part, the protective effect of plant extracts as antiplatelet agents. PMID:24190032

  13. File list: Unc.Adp.05.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  19. File list: Unc.Adp.05.Unclassified.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  20. An adpA homologue in Streptomyces avermitilis is involved in regulation of morphogenesis and melanogenesis

    Institute of Scientific and Technical Information of China (English)

    ZHAO JinLei; WEN Ying; CHEN Zhi; SONG Yuan; LI JiLun

    2007-01-01

    In Streptomyces griseus, AdpA, the key transcriptional activator in the A-factor regulatory cascade, switches on the transcription of multiple genes required for secondary metabolism and morphological differentiation. Streptomyces avermitilis also contains an ortholog of adpA, which is named adpA-a. To clarify the in vivo function of adpA-a, an adpA-a-disrupted strain was constructed by double crossover recombination. No difference in avermectin production was found between the adpA-a-disruptant and the wild-type strain. However, this disruptant neither formed spores nor produced melanin and its phenotype was restored to the original wild-type by a single copy of the adpA-a gene integrated into the chromosome. This report shows that adpA-a is involved in regulation of morphological differentiation and melanin production in S. avermitilis.

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  3. File list: His.Adp.20.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.White_adipocytes mm9 Histone Adipocyte White adipocytes SRX800009 ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.20.AllAg.White_adipocytes.bed ...

  4. File list: His.Adp.50.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.White_adipocytes mm9 Histone Adipocyte White adipocytes SRX800009 ...http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.50.AllAg.White_adipocytes.bed ...

  5. Impact of Dabigatran versus Phenprocoumon on ADP Induced Platelet Aggregation in Patients with Atrial Fibrillation with or without Concomitant Clopidogrel Therapy (the Dabi-ADP-1 and Dabi-ADP-2 Trials

    Directory of Open Access Journals (Sweden)

    Amadea M. Martischnig

    2015-01-01

    Full Text Available Background. A relevant number of patients receive triple therapy with clopidogrel, aspirin, and oral anticoagulation. Clopidogrel’s efficacy on ADP induced platelet function may be influenced by concomitant antithrombotic therapies. Data regarding the effect of dabigatran on platelet function is limited to in vitro studies and healthy individuals. Methods. The “Dabi-ADP-1” and “Dabi-ADP-2” trials randomized patients with atrial fibrillation to either dabigatran or phenprocoumon for a 2-week period. In Dabi-ADP-1 (n=70 patients with clopidogrel therapy were excluded and in Dabi-ADP-2 (n=46 patients had to be treated concomitantly with clopidogrel. The primary endpoint was ADP-induced platelet aggregation between dabigatran and phenprocoumon at 14 days. Secondary endpoints were ADPtest HS-, TRAP-, and COL-induced platelet aggregation. Results. There was no significant difference regarding the primary endpoint between both groups in either trial (Dabi-ADP-1: Dabigatran: 846 [650–983] AU × min versus phenprocoumon: 839 [666–1039] AU × min, P=0.90 and Dabi-ADP-2: 326 [268–462] versus 350 [214–535], P=0.70 or regarding the secondary endpoints, ADPtest HS-, TRAP-, and COL-induced platelet aggregation. Conclusion. Dabigatran as compared to phenprocoumon has no impact on ADP-induced platelet aggregation in atrial fibrillation patients neither with nor without concomitant clopidogrel therapy.

  6. Abiogenic photophosphorylation of ADP to ATP sensitized by flavoproteinoid microspheres.

    Science.gov (United States)

    Kolesnikov, Michael P; Telegina, Taisiya A; Lyudnikova, Tamara A; Kritsky, Mikhail S

    2008-06-01

    A model for abiogenic photophosphorylation of ADP by orthophosphate to yield ATP was studied. The model is based on the photochemical activity of flavoproteinoid microspheres that are formed by aggregation in an aqueous medium of products of thermal condensation of a glutamic acid, glycine and lysine mixture (8:3:1) and contain, along with amino acid polymers (proteinoids), abiogenic isoalloxazine (flavin) pigments. Irradiation of aqueous suspensions of microspheres with blue visible light or ultraviolet in the presence of ADP and orthophosphate resulted in ATP formation. The yield of ATP in aerated suspensions was 10-20% per one mol of starting ADP. Deaeration reduced the photophosphorylating activity of microspheres five to 10 times. Treatment of aerated microsphere suspensions with superoxide dismutase during irradiation partially suppressed ATP formation. Deaerated microspheres restored completely their photophosphorylating activity after addition of hydrogen peroxide to the suspension. The photophosphorylating activity of deaerated suspensions of flavoproteinoid microspheres was also recovered by introduction of Fe3+-cytochrome c, an electron acceptor alternative to oxygen. On the basis of the results obtained, a chemical mechanism of phosphorylation is proposed in which the free radical form of reduced flavin sensitizer (F1H*) and ADP are involved. PMID:18386156

  7. Characterization of archaeal group II chaperonin-ADP-metal fluoride complexes: implications that group II chaperonins operate as a "two-stroke engine".

    Science.gov (United States)

    Iizuka, Ryo; Yoshida, Takao; Ishii, Noriyuki; Zako, Tamotsu; Takahashi, Kazunobu; Maki, Kosuke; Inobe, Tomonao; Kuwajima, Kunihiro; Yohda, Masafumi

    2005-12-01

    Group II chaperonins, found in Archaea and in the eukaryotic cytosol, act independently of a cofactor corresponding to GroES of group I chaperonins. Instead, the helical protrusion at the tip of the apical domain forms a built-in lid of the central cavity. Although many studies on the lid's conformation have been carried out, the conformation in each step of the ATPase cycle remains obscure. To clarify this issue, we examined the effects of ADP-aluminum fluoride (AlFx) and ADP-beryllium fluoride (BeFx) complexes on alpha-chaperonin from the hyperthermophilic archaeum, Thermococcus sp. strain KS-1. Biochemical assays, electron microscopic observations, and small angle x-ray scattering measurements demonstrate that alpha-chaperonin incubated with ADP and BeFx exists in an asymmetric conformation; one ring is open, and the other is closed. The result indicates that alpha-chaperonin also shares the inherent functional asymmetry of bacterial and eukaryotic cytosolic chaperonins. Most interestingly, addition of ADP and BeFx induced alpha-chaperonin to encapsulate unfolded proteins in the closed ring but did not trigger their folding. Moreover, alpha-chaperonin incubated with ATP and AlFx or BeFx adopted a symmetric closed conformation, and its functional turnover was inhibited. These forms are supposed to be intermediates during the reaction cycle of group II chaperonins.

  8. 26 CFR 1.401(k)-2 - ADP test.

    Science.gov (United States)

    2010-04-01

    ...)(2)(iv) (as it appeared in the April 1, 2007, edition of 26 CFR part 1). (v) Distribution. Within 12... determined under § 1.401(k)-2(b)(2)(vi) (as it appeared in the April 1, 2007, edition of 26 CFR Part 1). (C... 26 Internal Revenue 5 2010-04-01 2010-04-01 false ADP test. 1.401(k)-2 Section 1.401(k)-2...

  9. A mitochondrial import receptor for the ADP/ATP carrier

    OpenAIRE

    Söllner, Thomas; Griffiths, Gareth; Pfanner, Nikolaus; Neupert, Walter

    1990-01-01

    We have identified a mitochondrial outer membrane protein of 72 kd (MOM72) that exhibits the properties of an import receptor for the ADP/ATP carrier (AAC), the most abundant mitochondrial protein. Monospecific antibodies and Fab fragments against MOM72 selectively inhibit import of AAC at the level of specific binding to the mitochondria. AAC bound to the mitochondrial surface is coprecipitated with antibodies against MOM72 after lysis of mitochondria with detergent. MOM72 thus has a complem...

  10. Arginine ADP-ribosyltransferase 1 promotes angiogenesis in colorectal cancer via the PI3K/Akt pathway.

    Science.gov (United States)

    Yang, Lian; Xiao, Ming; Li, Xian; Tang, Yi; Wang, Ya-Lan

    2016-03-01

    Arginine adenosine diphosphate (ADP)-ribosyl-transferase 1 (ART1) is known to play an important role in many physiological and pathological processes. Previous studies have demonstrated that ART1 promotes proliferation, invasion and metastasis in colon carcinoma. However, it was unclear whether ART1 is involved in angiogenesis in cases of colorectal cancer (CRC). In the present study, lentiviral vector‑mediated ART1‑cDNA or ART1-shRNA were transfected into LoVo cells, and the LoVo cells transfected with ART1-cDNA or ART1-shRNA were co-cultured with human umbilical vein endothelial cells (HUVECs) to determine the influence of ART1 on HUVECs. The proliferation, migration and angiogenesis of HUVECs were monitored using a cell counting kit-8 assay, a Transwell migration assay and immunohistochemical analysis in intrasplenic allograft tumors, respectively. Hypoxia‑inducible factor 1-α (HIF-1α), total (t-)Akt, phosphorylated (p-)Akt, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression levels were detected via western blot analysis. Our results revealed that HUVECs which were co-cultured with ART1-cDNA LoVo cells showed higher proliferation, migration and angiogenic abilities, but a reduction was noted in those cultured with ART1-shRNA LoVo cells; p-Akt, HIF-1α, VEGF and bFGF expression was increased in HUVECs cultured with ART1‑cDNA-transfected LoVo cells, but reduced in ART1-shRNA-transfected LoVo cells. In a mouse xenograft model, we noted that the tumor microvessel density (MVD) was significantly increased in intrasplenic transplanted ART1‑cDNA CT26 tumors but decreased in intrasplenic transplanted ART1‑shRNA tumors. These data suggest that ART1 promoted the expression of HIF-1α via the Akt pathway in tumor cells. It also upregulated VEGF and bFGF and enhanced angiogenesis in HUVECs. Thus, we suggest that ART1 plays an important role in the invasion of CRC cells and the metastasis of CRC.

  11. Structures of the human poly (ADP-ribose glycohydrolase catalytic domain confirm catalytic mechanism and explain inhibition by ADP-HPD derivatives.

    Directory of Open Access Journals (Sweden)

    Julie A Tucker

    Full Text Available Poly(ADP-ribose glycohydrolase (PARG is the only enzyme known to catalyse hydrolysis of the O-glycosidic linkages of ADP-ribose polymers, thereby reversing the effects of poly(ADP-ribose polymerases. PARG deficiency leads to cell death whilst PARG depletion causes sensitisation to certain DNA damaging agents, implicating PARG as a potential therapeutic target in several disease areas. Efforts to develop small molecule inhibitors of PARG activity have until recently been hampered by a lack of structural information on PARG. We have used a combination of bio-informatic and experimental approaches to engineer a crystallisable, catalytically active fragment of human PARG (hPARG. Here, we present high-resolution structures of the catalytic domain of hPARG in unliganded form and in complex with three inhibitors: ADP-ribose (ADPR, adenosine 5'-diphosphate (hydroxymethylpyrrolidinediol (ADP-HPD and 8-n-octyl-amino-ADP-HPD. Our structures confirm conservation of overall fold amongst mammalian PARG glycohydrolase domains, whilst revealing additional flexible regions in the catalytic site. These new structures rationalise a body of published mutational data and the reported structure-activity relationship for ADP-HPD based PARG inhibitors. In addition, we have developed and used biochemical, isothermal titration calorimetry and surface plasmon resonance assays to characterise the binding of inhibitors to our PARG protein, thus providing a starting point for the design of new inhibitors.

  12. Insights into the Mechanism of ADP Action on Flagellar Motility Derived from Studies on Bull Sperm

    OpenAIRE

    Lesich, Kathleen A.; Pelle, Dominic W.; Lindemann, Charles B.

    2008-01-01

    Adenosine diphosphate (ADP) is known to have interesting effects on flagellar motility. Permeabilized and reactivated bull sperm exhibit a marked reduction in beating frequency and a greatly increased beat amplitude in the presence of 1–4 mM ADP. In this study we examined the force production of sperm reactivated with 0.1 mM ATP with and without 1 mM ADP and found that there is little or no resulting change in the stalling force produced by a bull sperm flagella in response to ADP. Because bu...

  13. Mint3/X11γ Is an ADP-Ribosylation Factor-dependent Adaptor that Regulates the Traffic of the Alzheimer's Precursor Protein from the Trans-Golgi Network

    OpenAIRE

    Shrivastava-Ranjan, Punya; Faundez, Victor; Fang, Guofu; Rees, Howard; Lah, James J.; Allan I Levey; Kahn, Richard A.

    2008-01-01

    β-Amyloid peptides (Aβ) are the major component of plaques in brains of Alzheimer's patients, and are they derived from the proteolytic processing of the β-amyloid precursor protein (APP). The movement of APP between organelles is highly regulated, and it is tightly connected to its processing by secretases. We proposed previously that transport of APP within the cell is mediated in part through its sorting into Mint/X11-containing carriers. To test our hypothesis, we purified APP-containing ...

  14. Chemical Bond Calculations of Crystal Growth of KDP and ADP

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A novel method was proposed to calculate the crystal morphology (or growth habit) on the basis of chemical bond analysis. All constituent chemical bonds were distinguished as relevant and independent bonds according to their variations during the crystallization process. By employing the current method, the influence of specific growth conditions on the crystal morphology can be considered in the structure analysis process. The ideal morphologies of both KDP (KH2PO4) and ADP (NH4H2PO4) crystals were calculated and compared with our obtained crystallites at room temperature, which validates the present calculation method very well.

  15. Import of ADP/ATP carrier into mitochondria

    OpenAIRE

    Steger, Heinrich F.; Söllner, Thomas; Kiebler, Michael; Dietmeier, Klaus A.; Pfaller, Rupert; Trülzsch, Konrad S.; Tropschug, Maximilian; Neupert, Walter; Pfanner, Nikolaus

    1990-01-01

    We have identified the yeast homologue of Neurospora crassa MOM72, the mitochondrial import receptor for the ADP/ATP carrier (AAC), by functional studies and by cDNA sequencing. Mitochondria of a yeast mutant in which the gene for MOM72 was disrupted were impaired in specific binding and import of AAC. Unexpectedly, we found a residual, yet significant import of AAC into mitochondria lacking MOM72 that occurred via the receptor MOM19. We conclude that both MOM72 and MOM19 can direct AAC into ...

  16. Topographic study of the ADP/ATP transport protein. Localization of ADP and atractyloside fixation sites. Identification of the antigenic domains

    International Nuclear Information System (INIS)

    The objectives of this research thesis were: to determine the intramolecular localisation of binding sites of atractyloside and adenine-nucleotides; to determine whether antibodies obtained against the ADP/ATP carrier protein and isolated from beef heart mitochondria possess a reactivity specific to the organ or the species, where antigenic determinants are localized and whether there is conservation of the antigenic structure from one species to the other; to study how to follow and interpret conformational changes of the protein under the effect of ADP and inhibitors (carboxy-atractyloside or bongkrekic acid), and where the SH group unmasked by ADP and bongkrekic acid is localized

  17. Differential activities of cellular and viral macro domain proteins in binding of ADP-ribose metabolites.

    Science.gov (United States)

    Neuvonen, Maarit; Ahola, Tero

    2009-01-01

    Macro domain is a highly conserved protein domain found in both eukaryotes and prokaryotes. Macro domains are also encoded by a set of positive-strand RNA viruses that replicate in the cytoplasm of animal cells, including coronaviruses and alphaviruses. The functions of the macro domain are poorly understood, but it has been suggested to be an ADP-ribose-binding module. We have here characterized three novel human macro domain proteins that were found to reside either in the cytoplasm and nucleus [macro domain protein 2 (MDO2) and ganglioside-induced differentiation-associated protein 2] or in mitochondria [macro domain protein 1 (MDO1)], and compared them with viral macro domains from Semliki Forest virus, hepatitis E virus, and severe acute respiratory syndrome coronavirus, and with a yeast macro protein, Poa1p. MDO2 specifically bound monomeric ADP-ribose with a high affinity (K(d)=0.15 microM), but did not bind poly(ADP-ribose) efficiently. MDO2 also hydrolyzed ADP-ribose-1'' phosphate, resembling Poa1p in all these properties. Ganglioside-induced differentiation-associated protein 2 did not show affinity for ADP-ribose or its derivatives, but instead bound poly(A). MDO1 was generally active in these reactions, including poly(A) binding. Individual point mutations in MDO1 abolished monomeric ADP-ribose binding, but not poly(ADP-ribose) binding; in poly(ADP-ribose) binding assays, the monomer did not compete against polymer binding. The viral macro proteins bound poly(ADP-ribose) and poly(A), but had a low affinity for monomeric ADP-ribose. Thus, the viral proteins do not closely resemble any of the human proteins in their biochemical functions. The differential activity profiles of the human proteins implicate them in different cellular pathways, some of which may involve RNA rather than ADP-ribose derivatives.

  18. Structures of Mycobacterium Tuberculosis Folylpolyglutamate Synthase Complexed With ADP And AMPPCD

    Energy Technology Data Exchange (ETDEWEB)

    Young, P.G.; Smith, C.A.; Metcalf, P.; Baker, E.N.

    2009-05-28

    Folate derivatives are essential vitamins for cell growth and replication, primarily because of their central role in reactions of one-carbon metabolism. Folates require polyglutamation to be efficiently retained within the cell and folate-dependent enzymes have a higher affinity for the polyglutamylated forms of this cofactor. Polyglutamylation is dependent on the enzyme folylpolyglutamate synthetase (FPGS), which catalyzes the sequential addition of several glutamates to folate. FPGS is essential for the growth and survival of important bacterial species, including Mycobacterium tuberculosis, and is a potential drug target. Here, the crystal structures of M. tuberculosis FPGS in complex with ADP and AMPPCP are reported at 2.0 and 2.3 angstroms resolution, respectively. The structures reveal a deeply buried nucleotide-binding site, as in the Escherichia coli and Lactobacillus casei FPGS structures, and a long extended groove for the binding of folate substrates. Differences from the E. coli and L. casei FPGS structures are seen in the binding of a key divalent cation, the carbamylation state of an essential lysine side chain and the adoption of an 'open' position by the active-site beta5-alpha6 loop. These changes point to coordinated events that are associated with dihydropteroate/folate binding and the catalysis of the new amide bond with an incoming glutamate residue.

  19. Investigation of the interaction between the large and small subunits of potato ADP-glucose pyrophosphorylase.

    Directory of Open Access Journals (Sweden)

    Ibrahim Baris

    2009-10-01

    Full Text Available ADP-glucose pyrophosphorylase (AGPase, a key allosteric enzyme involved in higher plant starch biosynthesis, is composed of pairs of large (LS and small subunits (SS. Current evidence indicates that the two subunit types play distinct roles in enzyme function. Recently the heterotetrameric structure of potato AGPase has been modeled. In the current study, we have applied the molecular mechanics generalized born surface area (MM-GBSA method and identified critical amino acids of the potato AGPase LS and SS subunits that interact with each other during the native heterotetrameric structure formation. We have further shown the role of the LS amino acids in subunit-subunit interaction by yeast two-hybrid, bacterial complementation assay and native gel. Comparison of the computational results with the experiments has indicated that the backbone energy contribution (rather than the side chain energies of the interface residues is more important in identifying critical residues. We have found that lateral interaction of the LS-SS is much stronger than the longitudinal one, and it is mainly mediated by hydrophobic interactions. This study will not only enhance our understanding of the interaction between the SS and the LS of AGPase, but will also enable us to engineer proteins to obtain better assembled variants of AGPase which can be used for the improvement of plant yield.

  20. Correlation between increased platelet ADP aggregability and silent brain infarcts

    International Nuclear Information System (INIS)

    The purpose of this study was to investigate the correlation between platelet aggregability and silent brain infarcts. The study subjects were 445 people (264 men, 181 women; mean age, 53±14 years) with no neurologic signs, history of brain tumor, trauma, cerebrovascular disease, or antiplatelet medications. Adenosine diphosphate (ADP)-induced platelet aggregation was measured by the aggregation-size analytic method. Platelet aggregability was classified into 9 classes. The presence of headache/vertigo, hypertension, diabetes mellitus, hyperlipidemia, or smoking was elicited by questioning or blood sampling. A head MRI scan was performed, and if marked atherosclerosis or obvious stenosis in the intracranial vessels was detected, it was defined as a positive MR angiography (MRA) finding. Silent brain infarcts were detected in 26.3% of subjects. Hyperaggregability defined as that above class 6, 7, and 8 was present in 43.8%, 30.8%, and 15.7% of subjects, respectively. The risk factors for silent brain infarcts by multiple logistic regression analysis were aging, hypertension, positive MRA findings, and hyperaggregability. Platelet ADP hyperaggregability might be a risk factor for silent brain infarcts. (author)

  1. File list: His.Adp.10.Pan_lysine_crotonylation.AllCell [Chip-atlas[Archive

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  4. File list: ALL.Adp.20.AllAg.AllCell [Chip-atlas[Archive

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  1. File list: NoD.Adp.10.AllAg.Brown_preadipocytes [Chip-atlas[Archive

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  2. File list: NoD.Adp.20.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.AllAg.Brown_preadipocytes mm9 No description Adipocyte Brown preadipocyt...es http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.20.AllAg.Brown_preadipocytes.bed ...

  3. File list: His.Adp.20.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Brown_preadipocytes mm9 Histone Adipocyte Brown preadipocytes SRX3...RX341420,SRX341421,SRX341046 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.20.AllAg.Brown_preadipocytes.bed ...

  4. File list: InP.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.AllAg.Brown_preadipocytes mm9 Input control Adipocyte Brown preadipocyte...056,SRX341058,SRX478161 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.50.AllAg.Brown_preadipocytes.bed ...

  5. File list: Pol.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.Brown_preadipocytes mm9 RNA polymerase Adipocyte Brown preadipocyt...es SRX341031,SRX341032,SRX341029,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.50.AllAg.Brown_preadipocytes.bed ...

  6. File list: Oth.Adp.10.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Brown_preadipocytes mm9 TFs and others Adipocyte Brown preadipocyt...341763,SRX341767,SRX341419,SRX341028,SRX341766 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.10.AllAg.Brown_preadipocytes.bed ...

  7. File list: ALL.Adp.20.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Brown_preadipocytes mm9 All antigens Adipocyte Brown preadipocytes...RX341044,SRX341420,SRX341421,SRX341046,SRX478161,SRX341027 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.20.AllAg.Brown_preadipocytes.bed ...

  8. File list: Pol.Adp.10.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.Brown_preadipocytes mm9 RNA polymerase Adipocyte Brown preadipocyt...es SRX341031,SRX341032,SRX341029,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.10.AllAg.Brown_preadipocytes.bed ...

  9. File list: DNS.Adp.05.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.05.AllAg.Brown_preadipocytes mm9 DNase-seq Adipocyte Brown preadipocytes ht...tp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.05.AllAg.Brown_preadipocytes.bed ...

  10. File list: His.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Brown_preadipocytes mm9 Histone Adipocyte Brown preadipocytes SRX3...RX341421,SRX341046,SRX478160 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.50.AllAg.Brown_preadipocytes.bed ...

  11. File list: Pol.Adp.05.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.AllAg.Brown_preadipocytes mm9 RNA polymerase Adipocyte Brown preadipocyt...es SRX341031,SRX341032,SRX341029,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.05.AllAg.Brown_preadipocytes.bed ...

  12. File list: His.Adp.05.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Brown_preadipocytes mm9 Histone Adipocyte Brown preadipocytes SRX3...RX341421,SRX341039,SRX341040 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.05.AllAg.Brown_preadipocytes.bed ...

  13. File list: DNS.Adp.20.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.20.AllAg.Brown_preadipocytes mm9 DNase-seq Adipocyte Brown preadipocytes ht...tp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.20.AllAg.Brown_preadipocytes.bed ...

  14. File list: Oth.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.Brown_preadipocytes mm9 TFs and others Adipocyte Brown preadipocyt...341028,SRX341760,SRX341767,SRX341763,SRX341027 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.50.AllAg.Brown_preadipocytes.bed ...

  15. File list: InP.Adp.20.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.AllAg.Brown_preadipocytes mm9 Input control Adipocyte Brown preadipocyte...782,SRX341056,SRX478161 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.20.AllAg.Brown_preadipocytes.bed ...

  16. File list: ALL.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Brown_preadipocytes mm9 All antigens Adipocyte Brown preadipocytes...RX341420,SRX341421,SRX341046,SRX478161,SRX341027,SRX478160 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.50.AllAg.Brown_preadipocytes.bed ...

  17. File list: Oth.Adp.05.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Brown_preadipocytes mm9 TFs and others Adipocyte Brown preadipocyt...341766,SRX341418,SRX341023,SRX341419,SRX341767 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.AllAg.Brown_preadipocytes.bed ...

  18. File list: ALL.Adp.10.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Brown_preadipocytes mm9 All antigens Adipocyte Brown preadipocytes...RX341420,SRX478161,SRX478160,SRX341040,SRX341041,SRX341039 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.10.AllAg.Brown_preadipocytes.bed ...

  19. File list: DNS.Adp.50.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.50.AllAg.Brown_preadipocytes mm9 DNase-seq Adipocyte Brown preadipocytes ht...tp://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.50.AllAg.Brown_preadipocytes.bed ...

  20. File list: InP.Adp.10.AllAg.Brown_preadipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.10.AllAg.Brown_preadipocytes mm9 Input control Adipocyte Brown preadipocyte...058,SRX341056,SRX478161 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.10.AllAg.Brown_preadipocytes.bed ...

  1. File list: Oth.Adp.05.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Pre-adipocytes hg19 TFs and others Adipocyte Pre-adipocytes SRX760...968,SRX760970,SRX760967,SRX760971,SRX760969 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.05.AllAg.Pre-adipocytes.bed ...

  2. File list: His.Adp.10.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.AllAg.Pre-adipocytes hg19 Histone Adipocyte Pre-adipocytes SRX760965,SRX...760962,SRX760966,SRX760964,SRX760963 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.AllAg.Pre-adipocytes.bed ...

  3. File list: Pol.Adp.20.RNA_polymerase_III.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.RNA_polymerase_III.AllCell hg19 RNA polymerase RNA polymerase III Adipoc...yte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.20.RNA_polymerase_III.AllCell.bed ...

  4. File list: His.Adp.20.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Capan-1 hg19 Histone Adipocyte Capan-1 SRX825378,SRX825364,SRX8253...85,SRX825371 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.AllAg.Capan-1.bed ...

  5. File list: His.Adp.10.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.AllAg.Capan-2 hg19 Histone Adipocyte Capan-2 SRX825386,SRX825379,SRX8253...72,SRX825365 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.AllAg.Capan-2.bed ...

  6. File list: ALL.Adp.10.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Capan-2 hg19 All antigens Adipocyte Capan-2 SRX825386,SRX825379,SR...X825372,SRX825365 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.10.AllAg.Capan-2.bed ...

  7. File list: ALL.Adp.50.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Capan-1 hg19 All antigens Adipocyte Capan-1 SRX825378,SRX825364,SR...X825385,SRX825392,SRX825371 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.50.AllAg.Capan-1.bed ...

  8. File list: His.Adp.05.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Capan-1 hg19 Histone Adipocyte Capan-1 SRX825378,SRX825371,SRX8253...64,SRX825385 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.05.AllAg.Capan-1.bed ...

  9. File list: ALL.Adp.20.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Capan-2 hg19 All antigens Adipocyte Capan-2 SRX825386,SRX825379,SR...X825365,SRX825372 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.20.AllAg.Capan-2.bed ...

  10. File list: His.Adp.50.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Capan-1 hg19 Histone Adipocyte Capan-1 SRX825378,SRX825364,SRX8253...85,SRX825371 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.AllAg.Capan-1.bed ...

  11. File list: ALL.Adp.05.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Capan-2 hg19 All antigens Adipocyte Capan-2 SRX825386,SRX825379,SR...X825372,SRX825365 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Capan-2.bed ...

  12. File list: His.Adp.05.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Capan-2 hg19 Histone Adipocyte Capan-2 SRX825386,SRX825379,SRX8253...72,SRX825365 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.05.AllAg.Capan-2.bed ...

  13. File list: His.Adp.50.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Capan-2 hg19 Histone Adipocyte Capan-2 SRX825386,SRX825379,SRX8253...65,SRX825372 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.AllAg.Capan-2.bed ...

  14. File list: ALL.Adp.20.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Capan-1 hg19 All antigens Adipocyte Capan-1 SRX825378,SRX825364,SR...X825385,SRX825371,SRX825392 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.20.AllAg.Capan-1.bed ...

  15. File list: ALL.Adp.10.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Capan-1 hg19 All antigens Adipocyte Capan-1 SRX825378,SRX825371,SR...X825364,SRX825385,SRX825392 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.10.AllAg.Capan-1.bed ...

  16. File list: His.Adp.20.AllAg.Capan-2 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Capan-2 hg19 Histone Adipocyte Capan-2 SRX825386,SRX825379,SRX8253...65,SRX825372 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.AllAg.Capan-2.bed ...

  17. File list: His.Adp.10.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.AllAg.Capan-1 hg19 Histone Adipocyte Capan-1 SRX825378,SRX825371,SRX8253...64,SRX825385 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.AllAg.Capan-1.bed ...

  18. File list: ALL.Adp.05.AllAg.Capan-1 [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Capan-1 hg19 All antigens Adipocyte Capan-1 SRX825378,SRX825371,SR...X825364,SRX825385,SRX825392 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Capan-1.bed ...

  19. File list: ALL.Adp.05.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Adipose_stromal_cell hg19 All antigens Adipocyte Adipose stromal c...019496,SRX019511,SRX019518,SRX019504,SRX019497,SRX019503 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Adipose_stromal_cell.bed ...

  20. File list: Pol.Adp.10.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.Adipose_stromal_cell hg19 RNA polymerase Adipocyte Adipose stromal... cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.10.AllAg.Adipose_stromal_cell.bed ...

  1. File list: His.Adp.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Adipoc...yte http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.10.Pan_lysine_acetylation.AllCell.bed ...

  2. File list: His.Adp.50.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.Pan_lysine_acetylation.AllCell.bed ...

  3. File list: His.Adp.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.Pan_lysine_acetylation.AllCell.bed ...

  4. File list: His.Adp.20.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.Pan_lysine_acetylation.AllCell mm9 Histone Pan lysine acetylation Adipoc...yte http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.20.Pan_lysine_acetylation.AllCell.bed ...

  5. File list: His.Adp.10.Pan_lysine_acetylation.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.Pan_lysine_acetylation.AllCell hg19 Histone Pan lysine acetylation Adipo...cyte http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.Pan_lysine_acetylation.AllCell.bed ...

  6. File list: Pol.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.AllAg.Adipose_Tissue,_White hg19 RNA polymerase Adipocyte Adipose Tissue..., White http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  7. File list: Pol.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.AllAg.Adipose_Tissue,_White hg19 RNA polymerase Adipocyte Adipose Tissue..., White http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  8. File list: NoD.Adp.50.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.50.AllAg.Adipose_Tissue hg19 No description Adipocyte Adipose Tissue SRX134...732 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.50.AllAg.Adipose_Tissue.bed ...

  9. File list: NoD.Adp.10.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.10.AllAg.Adipose_Tissue hg19 No description Adipocyte Adipose Tissue SRX134...732 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.10.AllAg.Adipose_Tissue.bed ...

  10. File list: His.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Adipose_Tissue,_White hg19 Histone Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  11. File list: ALL.Adp.05.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Adipose_Tissue hg19 All antigens Adipocyte Adipose Tissue SRX13473...2 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Adipose_Tissue.bed ...

  12. File list: Unc.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.05.AllAg.Adipose_Tissue,_White hg19 Unclassified Adipocyte Adipose Tissue, ...White http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  13. File list: ALL.Adp.20.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Adipose_Tissue hg19 All antigens Adipocyte Adipose Tissue SRX13473...2 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.20.AllAg.Adipose_Tissue.bed ...

  14. File list: Oth.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.Adipose_Tissue,_White hg19 TFs and others Adipocyte Adipose Tissue...SRX821810,SRX821806,SRX821809,SRX821817,SRX821816,SRX821807 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  15. File list: ALL.Adp.10.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Adipose_Tissue hg19 All antigens Adipocyte Adipose Tissue SRX13473...2 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.10.AllAg.Adipose_Tissue.bed ...

  16. File list: ALL.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Adipose_Tissue,_White hg19 All antigens Adipocyte Adipose Tissue, ...X821817,SRX821821,SRX821815,SRX821811,SRX821810,SRX821809 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  17. File list: ALL.Adp.50.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Adipose_Tissue hg19 All antigens Adipocyte Adipose Tissue SRX13473...2 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.50.AllAg.Adipose_Tissue.bed ...

  18. File list: Oth.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Adipose_Tissue,_White hg19 TFs and others Adipocyte Adipose Tissue...SRX821821,SRX821815,SRX821811,SRX821817,SRX821809,SRX821810 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  19. File list: DNS.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.10.AllAg.Adipose_Tissue,_White hg19 DNase-seq Adipocyte Adipose Tissue, Whi...te http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  20. File list: NoD.Adp.05.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.05.AllAg.Adipose_Tissue hg19 No description Adipocyte Adipose Tissue SRX134...732 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.05.AllAg.Adipose_Tissue.bed ...

  1. File list: NoD.Adp.20.AllAg.Adipose_Tissue [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.AllAg.Adipose_Tissue hg19 No description Adipocyte Adipose Tissue SRX134...732 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.20.AllAg.Adipose_Tissue.bed ...

  2. File list: Pol.Adp.10.RNA_Polymerase_III.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.RNA_Polymerase_III.AllCell mm9 RNA polymerase RNA Polymerase III Adipocy...te http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.10.RNA_Polymerase_III.AllCell.bed ...

  3. File list: InP.Adp.50.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.Input_control.AllCell mm9 Input control Input control Adipocyte SRX18587...27367 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.50.Input_control.AllCell.bed ...

  4. File list: InP.Adp.05.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.05.Input_control.AllCell hg19 Input control Input control Adipocyte SRX0194...p://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.05.Input_control.AllCell.bed ...

  5. File list: Pol.Adp.50.RNA_Polymerase_II.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.RNA_Polymerase_II.AllCell mm9 RNA polymerase RNA Polymerase II Adipocyte... SRX800011,SRX800010,SRX341031,SRX341032,SRX341029,SRX800016,SRX800017,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.50.RNA_Polymerase_II.AllCell.bed ...

  6. File list: Pol.Adp.05.RNA_Polymerase_II.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.RNA_Polymerase_II.AllCell mm9 RNA polymerase RNA Polymerase II Adipocyte... SRX800011,SRX800010,SRX800016,SRX800017,SRX341031,SRX341032,SRX341029,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.05.RNA_Polymerase_II.AllCell.bed ...

  7. File list: Pol.Adp.20.RNA_Polymerase_II.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.RNA_Polymerase_II.AllCell mm9 RNA polymerase RNA Polymerase II Adipocyte... SRX800011,SRX800010,SRX341031,SRX341032,SRX341029,SRX800016,SRX800017,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.20.RNA_Polymerase_II.AllCell.bed ...

  8. File list: Pol.Adp.10.RNA_Polymerase_II.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.RNA_Polymerase_II.AllCell mm9 RNA polymerase RNA Polymerase II Adipocyte... SRX800011,SRX800010,SRX800016,SRX341031,SRX341032,SRX341029,SRX800017,SRX341030 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.10.RNA_Polymerase_II.AllCell.bed ...

  9. File list: NoD.Adp.50.NA.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.50.NA.AllCell hg19 No description NA Adipocyte SRX134732,SRX031428,SRX08865...0,SRX031386,SRX056801,SRX031444,SRX312175,SRX056800,SRX088647,SRX312171 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.50.NA.AllCell.bed ...

  10. File list: NoD.Adp.20.NA.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.NA.AllCell hg19 No description NA Adipocyte SRX088650,SRX134732,SRX31217...5,SRX031428,SRX031386,SRX056801,SRX031444,SRX312171,SRX056800,SRX088647 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.20.NA.AllCell.bed ...

  11. File list: Unc.Adp.20.Unclassified.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.20.Unclassified.AllCell mm9 Unclassified Unclassified Adipocyte SRX978685,S...RX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.20.Unclassified.AllCell.bed ...

  12. File list: Unc.Adp.05.Unclassified.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.05.Unclassified.AllCell mm9 Unclassified Unclassified Adipocyte SRX978685,S...RX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.05.Unclassified.AllCell.bed ...

  13. File list: Oth.Adp.10.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Adipose_progenitor_cells mm9 TFs and others Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.10.AllAg.Adipose_progenitor_cells.bed ...

  14. File list: Unc.Adp.05.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.05.AllAg.Adipose_stromal_cell hg19 Unclassified Adipocyte Adipose stromal c...ell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.05.AllAg.Adipose_stromal_cell.bed ...

  15. File list: DNS.Adp.20.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.20.AllAg.Adipose_stromal_cell hg19 DNase-seq Adipocyte Adipose stromal cell... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.20.AllAg.Adipose_stromal_cell.bed ...

  16. File list: Unc.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.20.AllAg.Adipose_progenitor_cells mm9 Unclassified Adipocyte Adipose progen...itor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.20.AllAg.Adipose_progenitor_cells.bed ...

  17. File list: DNS.Adp.50.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.50.AllAg.Adipose_progenitor_cells mm9 DNase-seq Adipocyte Adipose progenito...r cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.50.AllAg.Adipose_progenitor_cells.bed ...

  18. File list: DNS.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.20.AllAg.Adipose_progenitor_cells mm9 DNase-seq Adipocyte Adipose progenito...r cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.20.AllAg.Adipose_progenitor_cells.bed ...

  19. File list: Unc.Adp.10.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.10.AllAg.Adipose_progenitor_cells mm9 Unclassified Adipocyte Adipose progen...itor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.10.AllAg.Adipose_progenitor_cells.bed ...

  20. File list: DNS.Adp.50.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.50.AllAg.Adipose_stromal_cell hg19 DNase-seq Adipocyte Adipose stromal cell... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.50.AllAg.Adipose_stromal_cell.bed ...

  1. File list: Unc.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.05.AllAg.Adipose_progenitor_cells mm9 Unclassified Adipocyte Adipose progen...itor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.05.AllAg.Adipose_progenitor_cells.bed ...

  2. File list: Unc.Adp.50.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.50.AllAg.Adipose_progenitor_cells mm9 Unclassified Adipocyte Adipose progen...itor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.50.AllAg.Adipose_progenitor_cells.bed ...

  3. File list: Oth.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Adipose_progenitor_cells mm9 TFs and others Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.AllAg.Adipose_progenitor_cells.bed ...

  4. File list: His.Adp.10.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.AllAg.Adipose_progenitor_cells mm9 Histone Adipocyte Adipose progenitor ...cells SRX127409,SRX127394,SRX127396,SRX127407,SRX127383,SRX127381 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.10.AllAg.Adipose_progenitor_cells.bed ...

  5. File list: His.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Adipose_progenitor_cells mm9 Histone Adipocyte Adipose progenitor ...cells SRX127394,SRX127409,SRX127396,SRX127407,SRX127381,SRX127383 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.20.AllAg.Adipose_progenitor_cells.bed ...

  6. File list: Pol.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.AllAg.Adipose_progenitor_cells mm9 RNA polymerase Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.20.AllAg.Adipose_progenitor_cells.bed ...

  7. File list: DNS.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.05.AllAg.Adipose_progenitor_cells mm9 DNase-seq Adipocyte Adipose progenito...r cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.05.AllAg.Adipose_progenitor_cells.bed ...

  8. File list: Oth.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.20.AllAg.Adipose_progenitor_cells mm9 TFs and others Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.20.AllAg.Adipose_progenitor_cells.bed ...

  9. File list: DNS.Adp.05.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.05.AllAg.Adipose_stromal_cell hg19 DNase-seq Adipocyte Adipose stromal cell... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.05.AllAg.Adipose_stromal_cell.bed ...

  10. File list: DNS.Adp.10.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.10.AllAg.Adipose_stromal_cell hg19 DNase-seq Adipocyte Adipose stromal cell... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.10.AllAg.Adipose_stromal_cell.bed ...

  11. File list: Pol.Adp.50.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.Adipose_progenitor_cells mm9 RNA polymerase Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.50.AllAg.Adipose_progenitor_cells.bed ...

  12. File list: His.Adp.50.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Adipose_stromal_cell hg19 Histone Adipocyte Adipose stromal cell S...15,SRX019508,SRX019494 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.AllAg.Adipose_stromal_cell.bed ...

  13. File list: His.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Adipose_progenitor_cells mm9 Histone Adipocyte Adipose progenitor ...cells SRX127409,SRX127407,SRX127394,SRX127396,SRX127383,SRX127381 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/His.Adp.05.AllAg.Adipose_progenitor_cells.bed ...

  14. File list: Pol.Adp.50.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.Adipose_stromal_cell hg19 RNA polymerase Adipocyte Adipose stromal... cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.50.AllAg.Adipose_stromal_cell.bed ...

  15. File list: Pol.Adp.20.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.AllAg.Adipose_stromal_cell hg19 RNA polymerase Adipocyte Adipose stromal... cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.20.AllAg.Adipose_stromal_cell.bed ...

  16. File list: Pol.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.AllAg.Adipose_progenitor_cells mm9 RNA polymerase Adipocyte Adipose prog...enitor cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.05.AllAg.Adipose_progenitor_cells.bed ...

  17. File list: DNS.Adp.10.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.10.AllAg.Adipose_progenitor_cells mm9 DNase-seq Adipocyte Adipose progenito...r cells http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/DNS.Adp.10.AllAg.Adipose_progenitor_cells.bed ...

  18. File list: Unc.Adp.10.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.10.AllAg.Adipose_stromal_cell hg19 Unclassified Adipocyte Adipose stromal c...ell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.10.AllAg.Adipose_stromal_cell.bed ...

  19. File list: Unc.Adp.50.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.50.AllAg.Adipose_stromal_cell hg19 Unclassified Adipocyte Adipose stromal c...ell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.50.AllAg.Adipose_stromal_cell.bed ...

  20. File list: Pol.Adp.05.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.AllAg.Adipocytes hg19 RNA polymerase Adipocyte Adipocytes SRX682086,SRX6...82084,SRX682083,SRX682085 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.05.AllAg.Adipocytes.bed ...

  1. File list: Oth.Adp.10.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Adipocytes hg19 TFs and others Adipocyte Adipocytes SRX027402,SRX0...27403,SRX027401 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.10.AllAg.Adipocytes.bed ...

  2. File list: His.Adp.05.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Pre-adipocytes hg19 Histone Adipocyte Pre-adipocytes SRX760966,SRX...760963,SRX760964,SRX760965,SRX760962 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.05.AllAg.Pre-adipocytes.bed ...

  3. File list: ALL.Adp.20.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Adipocytes hg19 All antigens Adipocyte Adipocytes SRX027402,SRX682...084,SRX682086,SRX682083,SRX682085,SRX027401,SRX027400,SRX027403,SRX027404 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.20.AllAg.Adipocytes.bed ...

  4. File list: Pol.Adp.20.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.AllAg.Adipocytes hg19 RNA polymerase Adipocyte Adipocytes SRX682084,SRX6...82086,SRX682083,SRX682085 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.20.AllAg.Adipocytes.bed ...

  5. File list: Oth.Adp.50.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.Pre-adipocytes hg19 TFs and others Adipocyte Pre-adipocytes SRX760...967,SRX760968,SRX760971,SRX760969,SRX760970 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.50.AllAg.Pre-adipocytes.bed ...

  6. File list: His.Adp.20.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Pre-adipocytes hg19 Histone Adipocyte Pre-adipocytes SRX760962,SRX...760966,SRX760963,SRX760965,SRX760964 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.AllAg.Pre-adipocytes.bed ...

  7. File list: ALL.Adp.50.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Adipocytes hg19 All antigens Adipocyte Adipocytes SRX682084,SRX682...086,SRX027402,SRX682085,SRX682083,SRX027401,SRX027400,SRX027404,SRX027403 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.50.AllAg.Adipocytes.bed ...

  8. File list: ALL.Adp.10.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Adipocytes hg19 All antigens Adipocyte Adipocytes SRX027402,SRX682...086,SRX682084,SRX027400,SRX682085,SRX682083,SRX027403,SRX027401,SRX027404 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.10.AllAg.Adipocytes.bed ...

  9. File list: His.Adp.50.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Pre-adipocytes hg19 Histone Adipocyte Pre-adipocytes SRX760966,SRX...760964,SRX760963,SRX760965,SRX760962 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.AllAg.Pre-adipocytes.bed ...

  10. File list: Pol.Adp.10.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.Adipocytes hg19 RNA polymerase Adipocyte Adipocytes SRX682086,SRX6...82084,SRX682085,SRX682083 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.10.AllAg.Adipocytes.bed ...

  11. File list: Oth.Adp.05.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Adipocytes hg19 TFs and others Adipocyte Adipocytes SRX027402,SRX0...27403,SRX027401 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.05.AllAg.Adipocytes.bed ...

  12. File list: Oth.Adp.50.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.Adipocytes hg19 TFs and others Adipocyte Adipocytes SRX027402,SRX0...27401,SRX027403 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.50.AllAg.Adipocytes.bed ...

  13. File list: Oth.Adp.20.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.20.AllAg.Pre-adipocytes hg19 TFs and others Adipocyte Pre-adipocytes SRX760...967,SRX760968,SRX760971,SRX760969,SRX760970 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.20.AllAg.Pre-adipocytes.bed ...

  14. File list: Oth.Adp.10.AllAg.Pre-adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Pre-adipocytes hg19 TFs and others Adipocyte Pre-adipocytes SRX760...968,SRX760967,SRX760971,SRX760969,SRX760970 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.10.AllAg.Pre-adipocytes.bed ...

  15. File list: ALL.Adp.05.AllAg.Adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Adipocytes hg19 All antigens Adipocyte Adipocytes SRX027402,SRX027...403,SRX027400,SRX682086,SRX682084,SRX682083,SRX682085,SRX027401,SRX027404 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Adipocytes.bed ...

  16. 7 CFR 277.18 - Establishment of an Automated Data Processing (ADP) and Information Retrieval System.

    Science.gov (United States)

    2010-01-01

    ...) and Information Retrieval System. 277.18 Section 277.18 Agriculture Regulations of the Department of... Data Processing (ADP) and Information Retrieval System. (a) Scope and application. This section... costs of planning, design, development or installation of ADP and information retrieval systems if...

  17. File list: ALL.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Brown_adipocytes mm9 All antigens Adipocyte Brown adipocytes SRX80...X800019,SRX185797,SRX478163,SRX478162 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.50.AllAg.Brown_adipocytes.bed ...

  18. File list: ALL.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Brown_adipocytes mm9 All antigens Adipocyte Brown adipocytes SRX80...X185879,SRX978689,SRX978688,SRX478162 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.05.AllAg.Brown_adipocytes.bed ...

  19. File list: Unc.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.20.AllAg.Brown_adipocytes mm9 Unclassified Adipocyte Brown adipocytes SRX97...8685,SRX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.20.AllAg.Brown_adipocytes.bed ...

  20. File list: NoD.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.50.AllAg.Brown_adipocytes mm9 No description Adipocyte Brown adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.50.AllAg.Brown_adipocytes.bed ...

  1. File list: Oth.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.Brown_adipocytes mm9 TFs and others Adipocyte Brown adipocytes SRX...RX978688,SRX800015,SRX800014,SRX800018,SRX800019 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.50.AllAg.Brown_adipocytes.bed ...

  2. File list: NoD.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.05.AllAg.Brown_adipocytes mm9 No description Adipocyte Brown adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.05.AllAg.Brown_adipocytes.bed ...

  3. File list: Pol.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.20.AllAg.Brown_adipocytes mm9 RNA polymerase Adipocyte Brown adipocytes SRX...800010,SRX800016,SRX800017 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.20.AllAg.Brown_adipocytes.bed ...

  4. File list: Unc.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.10.AllAg.Brown_adipocytes mm9 Unclassified Adipocyte Brown adipocytes SRX97...8685,SRX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.10.AllAg.Brown_adipocytes.bed ...

  5. File list: Oth.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.20.AllAg.Brown_adipocytes mm9 TFs and others Adipocyte Brown adipocytes SRX...RX978689,SRX800015,SRX800014,SRX800018,SRX800019 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.20.AllAg.Brown_adipocytes.bed ...

  6. File list: NoD.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.10.AllAg.Brown_adipocytes mm9 No description Adipocyte Brown adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.10.AllAg.Brown_adipocytes.bed ...

  7. File list: NoD.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.AllAg.Brown_adipocytes mm9 No description Adipocyte Brown adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.20.AllAg.Brown_adipocytes.bed ...

  8. File list: InP.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.AllAg.Brown_adipocytes mm9 Input control Adipocyte Brown adipocytes SRX1...85879,SRX143805,SRX478163 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.50.AllAg.Brown_adipocytes.bed ...

  9. File list: Pol.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.Brown_adipocytes mm9 RNA polymerase Adipocyte Brown adipocytes SRX...800010,SRX800016,SRX800017 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.50.AllAg.Brown_adipocytes.bed ...

  10. File list: Oth.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.10.AllAg.Brown_adipocytes mm9 TFs and others Adipocyte Brown adipocytes SRX...RX800014,SRX978690,SRX978689,SRX978688,SRX800019 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.10.AllAg.Brown_adipocytes.bed ...

  11. File list: InP.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.10.AllAg.Brown_adipocytes mm9 Input control Adipocyte Brown adipocytes SRX1...43805,SRX185879,SRX478163 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.10.AllAg.Brown_adipocytes.bed ...

  12. File list: ALL.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.20.AllAg.Brown_adipocytes mm9 All antigens Adipocyte Brown adipocytes SRX80...X800018,SRX800019,SRX185797,SRX478162 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.20.AllAg.Brown_adipocytes.bed ...

  13. File list: ALL.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Brown_adipocytes mm9 All antigens Adipocyte Brown adipocytes SRX80...X978688,SRX800019,SRX478163,SRX478162 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/ALL.Adp.10.AllAg.Brown_adipocytes.bed ...

  14. File list: Unc.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.05.AllAg.Brown_adipocytes mm9 Unclassified Adipocyte Brown adipocytes SRX97...8685,SRX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.05.AllAg.Brown_adipocytes.bed ...

  15. File list: Pol.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.05.AllAg.Brown_adipocytes mm9 RNA polymerase Adipocyte Brown adipocytes SRX...800010,SRX800016,SRX800017 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.05.AllAg.Brown_adipocytes.bed ...

  16. File list: Oth.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Brown_adipocytes mm9 TFs and others Adipocyte Brown adipocytes SRX...RX800019,SRX978691,SRX978690,SRX978689,SRX978688 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.AllAg.Brown_adipocytes.bed ...

  17. File list: InP.Adp.20.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.AllAg.Brown_adipocytes mm9 Input control Adipocyte Brown adipocytes SRX1...85879,SRX143805,SRX478163 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.20.AllAg.Brown_adipocytes.bed ...

  18. File list: InP.Adp.05.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.05.AllAg.Brown_adipocytes mm9 Input control Adipocyte Brown adipocytes SRX4...78163,SRX143805,SRX185879 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.05.AllAg.Brown_adipocytes.bed ...

  19. File list: Unc.Adp.50.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.50.AllAg.Brown_adipocytes mm9 Unclassified Adipocyte Brown adipocytes SRX97...8685,SRX800022 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Unc.Adp.50.AllAg.Brown_adipocytes.bed ...

  20. File list: Pol.Adp.10.AllAg.Brown_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.Brown_adipocytes mm9 RNA polymerase Adipocyte Brown adipocytes SRX...800010,SRX800016,SRX800017 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.10.AllAg.Brown_adipocytes.bed ...

  1. A SPECTROPHOTOMETRIC ASSAY TO MEASURE RUBISCO ACTIVASE ACTIVATION ACTIVITY UNDER VARYING ATP:ADP RATIOS

    Science.gov (United States)

    The ratio of ATP to ADP in the stroma is an important regulatory mechanism for controlling the activation state of Rubisco via Rubisco activase (activase). Understanding the response of activase to a varying ATP:ADP ratio should reveal insights into the regulation of photosynthesis. However, the cur...

  2. File list: Oth.Adp.05.AllAg.SGBS [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.SGBS hg19 TFs and others Adipocyte SGBS SRX813768,SRX813771,SRX813...X813775,SRX032891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.05.AllAg.SGBS.bed ...

  3. File list: DNS.Adp.50.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.50.AllAg.Fetal_Heart hg19 DNase-seq Adipocyte Fetal Heart SRX040387,SRX0404...0390 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.50.AllAg.Fetal_Heart.bed ...

  4. REDUCED THROMBOGENICITY OF VASCULAR PROSTHESES BY COATING WITH ADP-ASE

    NARCIS (Netherlands)

    VANDERLEI, B; ROBINSON, PH; BAKKER, WW; Bartels, H.

    1992-01-01

    In this pilot study ADP-ase coated polyurethane (PL) vascular prostheses and noncoated (control) PU vascular prostheses (all vascular prostheses: ID 1.5 mm, length 1,5 cm) were implanted into the carotid artery of the rabbit to test wheter ADP-ase might function as an adequate anti-thrombogenic coat

  5. File list: InP.Adp.10.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.10.Input_control.AllCell hg19 Input control Input control Adipocyte SRX0194...p://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.10.Input_control.AllCell.bed ...

  6. File list: InP.Adp.20.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.Input_control.AllCell hg19 Input control Input control Adipocyte SRX0194...p://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.20.Input_control.AllCell.bed ...

  7. File list: InP.Adp.20.AllAg.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.AllAg.AllCell hg19 Input control Adipocyte SRX019491,SRX660092,SRX660091...,SRX1272789,SRX1272801,SRX032892,SRX196110,SRX469459,SRX469457,SRX825392,SRX027404 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.20.AllAg.AllCell.bed ...

  8. File list: InP.Adp.20.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.Input_control.AllCell mm9 Input control Input control Adipocyte SRX99775...78161 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.20.Input_control.AllCell.bed ...

  9. File list: InP.Adp.10.AllAg.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.10.AllAg.AllCell hg19 Input control Adipocyte SRX019491,SRX660092,SRX127280...1,SRX196110,SRX660091,SRX032892,SRX825392,SRX1272789,SRX469459,SRX469457,SRX027404 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.10.AllAg.AllCell.bed ...

  10. File list: InP.Adp.10.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.10.Input_control.AllCell mm9 Input control Input control Adipocyte SRX99775...78161 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.10.Input_control.AllCell.bed ...

  11. File list: InP.Adp.05.Input_control.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.05.Input_control.AllCell mm9 Input control Input control Adipocyte SRX99775...27370 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.05.Input_control.AllCell.bed ...

  12. File list: NoD.Adp.05.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.05.AllAg.Fetal_Heart hg19 No description Adipocyte Fetal Heart SRX088650,SR...X088647,SRX056801,SRX031428,SRX031386,SRX031444,SRX056800 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.05.AllAg.Fetal_Heart.bed ...

  13. File list: His.Adp.20.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Fetal_Heart hg19 Histone Adipocyte Fetal Heart SRX860893,SRX860898...,SRX860890,SRX860889,SRX860894,SRX860892,SRX860896,SRX860895,SRX860891,SRX860897 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.AllAg.Fetal_Heart.bed ...

  14. File list: NoD.Adp.20.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.AllAg.Fetal_Heart hg19 No description Adipocyte Fetal Heart SRX088650,SR...X031428,SRX031386,SRX056801,SRX031444,SRX056800,SRX088647 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.20.AllAg.Fetal_Heart.bed ...

  15. File list: His.Adp.05.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.05.AllAg.Fetal_Heart hg19 Histone Adipocyte Fetal Heart SRX860893,SRX860894...,SRX860897,SRX860898,SRX860892,SRX860890,SRX860889,SRX860896,SRX860895,SRX860891 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.05.AllAg.Fetal_Heart.bed ...

  16. File list: DNS.Adp.20.AllAg.Fetal_Heart [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.20.AllAg.Fetal_Heart hg19 DNase-seq Adipocyte Fetal Heart SRX040387,SRX0404...0390 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.20.AllAg.Fetal_Heart.bed ...

  17. File list: Pol.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.Adipose_Tissue,_White hg19 RNA polymerase Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  18. File list: DNS.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.50.AllAg.Adipose_Tissue,_White hg19 DNase-seq Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  19. File list: NoD.Adp.05.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.05.AllAg.White_adipocytes mm9 No description Adipocyte White adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.05.AllAg.White_adipocytes.bed ...

  20. File list: Oth.Adp.05.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.White_adipocytes mm9 TFs and others Adipocyte White adipocytes SRX... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.AllAg.White_adipocytes.bed ...

  1. File list: Unc.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.50.AllAg.Adipose_Tissue,_White hg19 Unclassified Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  2. File list: ALL.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.50.AllAg.Adipose_Tissue,_White hg19 All antigens Adipocyte Adipose Tissue, White...X821810,SRX821806,SRX821809,SRX821817,SRX821816,SRX821807 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  3. File list: Unc.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.20.AllAg.Adipose_Tissue,_White hg19 Unclassified Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  4. File list: Oth.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.AllAg.Adipose_Tissue,_White hg19 TFs and others Adipocyte Adipose Tissue, White...SRX821815,SRX821821,SRX821816,SRX821809,SRX821817,SRX821810 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  5. File list: His.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.50.AllAg.Adipose_Tissue,_White hg19 Histone Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  6. File list: Unc.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Unc.Adp.10.AllAg.Adipose_Tissue,_White hg19 Unclassified Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Unc.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  7. File list: DNS.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.05.AllAg.Adipose_Tissue,_White hg19 DNase-seq Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  8. File list: Pol.Adp.10.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.10.AllAg.White_adipocytes mm9 RNA polymerase Adipocyte White adipocytes SRX...800011 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.10.AllAg.White_adipocytes.bed ...

  9. File list: Oth.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.20.AllAg.Adipose_Tissue,_White hg19 TFs and others Adipocyte Adipose Tissue, White...SRX821817,SRX821821,SRX821815,SRX821811,SRX821810,SRX821809 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Oth.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  10. File list: His.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.20.AllAg.Adipose_Tissue,_White hg19 Histone Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  11. File list: NoD.Adp.50.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.50.AllAg.White_adipocytes mm9 No description Adipocyte White adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.50.AllAg.White_adipocytes.bed ...

  12. File list: DNS.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available DNS.Adp.20.AllAg.Adipose_Tissue,_White hg19 DNase-seq Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/DNS.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  13. File list: InP.Adp.05.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.05.AllAg.White_adipocytes mm9 Input control Adipocyte White adipocytes SRX9...97757,SRX821800,SRX821801,SRX268023,SRX821799 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.05.AllAg.White_adipocytes.bed ...

  14. File list: Pol.Adp.50.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.White_adipocytes mm9 RNA polymerase Adipocyte White adipocytes SRX...800011 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Pol.Adp.50.AllAg.White_adipocytes.bed ...

  15. File list: ALL.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.10.AllAg.Adipose_Tissue,_White hg19 All antigens Adipocyte Adipose Tissue, White...X821821,SRX821815,SRX821811,SRX821817,SRX821809,SRX821810 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  16. File list: His.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available His.Adp.10.AllAg.Adipose_Tissue,_White hg19 Histone Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/His.Adp.10.AllAg.Adipose_Tissue,_White.bed ...

  17. File list: ALL.Adp.05.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available ALL.Adp.05.AllAg.Adipose_Tissue,_White hg19 All antigens Adipocyte Adipose Tissue, White...X821815,SRX821821,SRX821816,SRX821809,SRX821817,SRX821810 http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/ALL.Adp.05.AllAg.Adipose_Tissue,_White.bed ...

  18. File list: NoD.Adp.20.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.20.AllAg.White_adipocytes mm9 No description Adipocyte White adipocytes htt...p://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/NoD.Adp.20.AllAg.White_adipocytes.bed ...

  19. File list: InP.Adp.50.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.AllAg.White_adipocytes mm9 Input control Adipocyte White adipocytes SRX2...68023,SRX997757,SRX821800,SRX821801,SRX821799 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.50.AllAg.White_adipocytes.bed ...

  20. File list: Oth.Adp.50.AllAg.White_adipocytes [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.AllAg.White_adipocytes mm9 TFs and others Adipocyte White adipocytes SRX... http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.50.AllAg.White_adipocytes.bed ...

  1. File list: Pol.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Pol.Adp.50.AllAg.Adipose_Tissue,_White hg19 RNA polymerase Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/Pol.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  2. Molecular and biochemical characterization of the ADP-dependent phosphofructokinase from the hyperthermophilic archaeon Pyrococcus furiosus.

    Science.gov (United States)

    Tuininga, J E; Verhees, C H; van der Oost, J; Kengen, S W; Stams, A J; de Vos, W M

    1999-07-23

    Pyrococcus furiosus uses a modified Embden-Meyerhof pathway involving two ADP-dependent kinases. Using the N-terminal amino acid sequence of the previously purified ADP-dependent glucokinase, the corresponding gene as well as a related open reading frame were detected in the genome of P. furiosus. Both genes were successfully cloned and expressed in Escherichia coli, yielding highly thermoactive ADP-dependent glucokinase and phosphofructokinase. The deduced amino acid sequences of both kinases were 21.1% identical but did not reveal significant homology with those of other known sugar kinases. The ADP-dependent phosphofructokinase was purified and characterized. The oxygen-stable protein had a native molecular mass of approximately 180 kDa and was composed of four identical 52-kDa subunits. It had a specific activity of 88 units/mg at 50 degrees C and a pH optimum of 6.5. As phosphoryl group donor, ADP could be replaced by GDP, ATP, and GTP to a limited extent. The K(m) values for fructose 6-phosphate and ADP were 2.3 and 0.11 mM, respectively. The phosphofructokinase did not catalyze the reverse reaction, nor was it regulated by any of the known allosteric modulators of ATP-dependent phosphofructokinases. ATP and AMP were identified as competitive inhibitors of the phosphofructokinase, raising the K(m) for ADP to 0.34 and 0.41 mM, respectively. PMID:10409652

  3. Structure of the Escherichia coli heptosyltransferase WaaC: binary complexes with ADP and ADP-2-deoxy-2-fluoro heptose.

    Science.gov (United States)

    Grizot, Sylvestre; Salem, Michèle; Vongsouthi, Vanida; Durand, Lionel; Moreau, François; Dohi, Hirofumi; Vincent, Stéphane; Escaich, Sonia; Ducruix, Arnaud

    2006-10-20

    Lipopolysaccharides constitute the outer leaflet of the outer membrane of Gram-negative bacteria and are therefore essential for cell growth and viability. The heptosyltransferase WaaC is a glycosyltransferase (GT) involved in the synthesis of the inner core region of LPS. It catalyzes the addition of the first L-glycero-D-manno-heptose (heptose) molecule to one 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) residue of the Kdo2-lipid A molecule. Heptose is an essential component of the LPS core domain; its absence results in a truncated lipopolysaccharide associated with the deep-rough phenotype causing a greater susceptibility to antibiotic and an attenuated virulence for pathogenic Gram-negative bacteria. Thus, WaaC represents a promising target in antibacterial drug design. Here, we report the structure of WaaC from the Escherichia coli pathogenic strain RS218 alone at 1.9 A resolution, and in complex with either ADP or the non-cleavable analog ADP-2-deoxy-2-fluoro-heptose of the sugar donor at 2.4 A resolution. WaaC adopts the GT-B fold in two domains, characteristic of one glycosyltransferase structural superfamily. The comparison of the three different structures shows that WaaC does not undergo a domain rotation, characteristic of the GT-B family, upon substrate binding, but allows the substrate analog and the reaction product to adopt remarkably distinct conformations inside the active site. In addition, both binary complexes offer a close view of the donor subsite and, together with results from site-directed mutagenesis studies, provide evidence for a model of the catalytic mechanism.

  4. Insights into the mechanism of ADP action on flagellar motility derived from studies on bull sperm.

    Science.gov (United States)

    Lesich, Kathleen A; Pelle, Dominic W; Lindemann, Charles B

    2008-07-01

    Adenosine diphosphate (ADP) is known to have interesting effects on flagellar motility. Permeabilized and reactivated bull sperm exhibit a marked reduction in beating frequency and a greatly increased beat amplitude in the presence of 1-4 mM ADP. In this study we examined the force production of sperm reactivated with 0.1 mM ATP with and without 1 mM ADP and found that there is little or no resulting change in the stalling force produced by a bull sperm flagella in response to ADP. Because bull sperm bend to a higher curvature after ADP treatment we explored the possibility that ADP-treated sperm flagella are more flexible. We measured the stiffness of 50 muM sodium vanadate treated bull sperm in the presence of 4 mM ADP, but found no change in the passive flagellar stiffness. When we analyzed the torque that develops in ADP-treated sperm at the point of beat reversal we found that the torque developed by the flagellum is significantly increased. Our torque estimates also allow us to calculate the transverse force (t-force) acting on the flagellum at the point of beat direction reversal. We find that the t-force at the switch-point of the beat is increased significantly in the ADP treated condition, averaging 0.7 +/- 0.29 nN/microm in 0.1 mM ATP and increasing to 2.9 +/- 1.2 nN/microm in 0.1 mM ATP plus 4 mM ADP. This suggests that ADP is exerting its effect on the beat by increasing the tenacity of dynein attachment at the B-subtubule. This could be a direct result of a regulatory effect of ADP on the binding affinity of dynein for the B-subtubule of the outer doublets. This result could also help to explain a number of previous experimental observations, as discussed. PMID:18375503

  5. Bacterial Vaginosis

    Science.gov (United States)

    ... 586. Related Content STDs during Pregnancy Fact Sheet Pregnancy and HIV, Viral Hepatitis, and STD Prevention Pelvic Inflammatory Disease ( ... Bacterial Vaginosis (BV) Chlamydia Gonorrhea Genital Herpes Hepatitis HIV/AIDS & STDs Human Papillomavirus ... STDs See Also Pregnancy Reproductive ...

  6. Bacterial Meningitis

    Science.gov (United States)

    ... Schedules Preteen & Teen Vaccines Meningococcal Disease Sepsis Bacterial Meningitis Recommend on Facebook Tweet Share Compartir On this ... serious disease. Laboratory Methods for the Diagnosis of Meningitis This manual summarizes laboratory methods used to isolate, ...

  7. The energetics of allosteric regulation of ADP release from myosin heads.

    Science.gov (United States)

    Jackson, Del R; Baker, Josh E

    2009-06-28

    Myosin molecules are involved in a wide range of transport and contractile activities in cells. A single myosin head functions through its ATPase reaction as a force generator and as a mechanosensor, and when two or more myosin heads work together in moving along an actin filament, the interplay between these mechanisms contributes to collective myosin behaviors. For example, the interplay between force-generating and force-sensing mechanisms coordinates the two heads of a myosin V molecule in its hand-over-hand processive stepping along an actin filament. In muscle, it contributes to the Fenn effect and smooth muscle latch. In both examples, a key force-sensing mechanism is the regulation of ADP release via interhead forces that are generated upon actin-myosin binding. Here we present a model describing the mechanism of allosteric regulation of ADP release from myosin heads as a change, DeltaDeltaG(-D), in the standard free energy for ADP release that results from the work, Deltamicro(mech), performed by that myosin head upon ADP release, or DeltaDeltaG(-D) = Deltamicro(mech). We show that this model is consistent with previous measurements for strain-dependent kinetics of ADP release in both myosin V and muscle myosin II. The model makes explicit the energetic cost of accelerating ADP release, showing that acceleration of ADP release during myosin V processivity requires approximately 4 kT of energy whereas the energetic cost for accelerating ADP release in a myosin II-based actin motility assay is only approximately 0.4 kT. The model also predicts that the acceleration of ADP release involves a dissipation of interhead forces. To test this prediction, we use an in vitro motility assay to show that the acceleration of ADP release from both smooth and skeletal muscle myosin II correlates with a decrease in interhead force. Our analyses provide clear energetic constraints for models of the allosteric regulation of ADP release and provide novel, testable insights

  8. Multiple forms of ADP-glucose pyrophosphorylase from tomato fruit

    Science.gov (United States)

    Chen, B. Y.; Janes, H. W.

    1997-01-01

    ADP-glucose pyrophosphorylase (AGP) was purified from tomato (Lycopersicon esculentum Mill.) fruit to apparent homogeneity. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis the enzyme migrated as two close bands with molecular weights of 50,000 and 51,000. Two-dimensional polyacrylamide gel electrophoresis analysis of the purified enzyme, however, revealed at least five major protein spots that could be distinguished by their slight differences in net charge and molecular weight. Whereas all of the spots were recognized by the antiserum raised against tomato fruit AGP holoenzyme, only three of them reacted strongly with antiserum raised against the potato tuber AGP large subunit, and the other two spots (with lower molecular weights) reacted specifically with antisera raised against spinach leaf AGP holoenzyme and the potato tuber AGP small subunit. The results suggest the existence of at least three isoforms of the AGP large subunit and two isoforms of the small subunit in tomato fruit in vivo. The native molecular mass of the enzyme determined by gel filtration was 220 +/- 10 kD, indicating a tetrameric structure for AGP from tomato fruit. The purified enzyme is very sensitive to 3-phosphoglycerate/inorganic phosphate regulation.

  9. Modification of ADP extinguishing powder by siliconization in spray drying

    Institute of Scientific and Technical Information of China (English)

    Xiaojing Zhang; Zhigang Shen; Chujiang Cai; Xiaozheng Yu; Jun Du; Yushan Xing; Shulin Ma

    2012-01-01

    Superfine spherical fire-extinguishing powder,ammonium dihydrogen phosphate (ADP,NH4H2PO4),was prepared by spray drying and modified in situ with methyl hydrogen silicone oil (MHSO) emulsion and the fluorinated surfactant FK-510.The influences of the MHSO mass ratio on the hydrophobicity,surface composition,surface morphology,dispersion and particle-size distribution of the NH4H2PO4 were studied,and the influence of the drying air temperature on the decomposition of the NH4H2PO4 was also researched.The results indicate that the MHSO and FK-510 congregate on the particle surfaces and then form a hydrophobic shell.This shell improves the particle hydrophobicity and leads to a fine dispersion of the particles.During the process of preparing the precursor solution,3 wt% (based on the weight of NH4H2PO4) was chosen as the optimum value of the MHSO mass ratio.During the spray drying,a low absolute humidity of the air should be maintained,and it is very important to keep the exit-air temperature below 100℃ to avoid decomposition.

  10. An affinity matrix for the purification of poly(ADP-ribose) glycohydrolase.

    OpenAIRE

    Thomassin, H; Jacobson, M K; Guay, J; Verreault, A; Aboul-Ela, N; Menard, L.; Poirier, G G

    1990-01-01

    The preparation of quantities of poly(ADP-ribose) glycohydrolase sufficient for detailed structural and enzymatic characterizations has been difficult due to the very low tissue content of the enzyme and its lability in late stages of purification. To date, the only purification of this enzyme to apparent homogeneity has involved a procedure requiring 6 column chromatographic steps. Described here is the preparation of an affinity matrix which consists of ADP-ribose polymers bound to dihydrox...

  11. Inhibition of Nuclear Receptor Signalling by Poly(ADP-Ribose) Polymerase

    OpenAIRE

    Miyamoto, Takahide; Kakizawa, Tomoko; Hashizume, Kiyoshi

    1999-01-01

    Mammalian poly(ADP-ribose) polymerase (PARP) is a nuclear chromatin-associated protein with a molecular mass of 114 kDa that catalyzes the transfer of ADP-ribose units from NAD+ to nuclear proteins that are located within chromatin. We report here the identification of a novel property of PARP as a modulator of nuclear receptor signalling. PARP bound directly to retinoid X receptors (RXR) and repressed ligand-dependent transcriptional activities mediated by heterodimers of RXR and thyroid hor...

  12. File list: Oth.Adp.50.Kmt2d.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.50.Kmt2d.AllCell mm9 TFs and others Kmt2d Adipocyte SRX339719,SRX339718,SRX...339717,SRX339721,SRX339722,SRX341764,SRX341418,SRX339720,SRX341766,SRX341765,SRX341419,SRX341767 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.50.Kmt2d.AllCell.bed ...

  13. File list: Oth.Adp.05.Kmt2d.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available Oth.Adp.05.Kmt2d.AllCell mm9 TFs and others Kmt2d Adipocyte SRX339719,SRX339722,SRX...339717,SRX339718,SRX339721,SRX341764,SRX341765,SRX339720,SRX341766,SRX341418,SRX341419,SRX341767 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/Oth.Adp.05.Kmt2d.AllCell.bed ...

  14. Analytical Design Package (ADP2): A computer aided engineering tool for aircraft transparency design

    Science.gov (United States)

    Wuerer, J. E.; Gran, M.; Held, T. W.

    1994-01-01

    The Analytical Design Package (ADP2) is being developed as a part of the Air Force Frameless Transparency Program (FTP). ADP2 is an integrated design tool consisting of existing analysis codes and Computer Aided Engineering (CAE) software. The objective of the ADP2 is to develop and confirm an integrated design methodology for frameless transparencies, related aircraft interfaces, and their corresponding tooling. The application of this methodology will generate high confidence for achieving a qualified part prior to mold fabrication. ADP2 is a customized integration of analysis codes, CAE software, and material databases. The primary CAE integration tool for the ADP2 is P3/PATRAN, a commercial-off-the-shelf (COTS) software tool. The open architecture of P3/PATRAN allows customized installations with different applications modules for specific site requirements. Integration of material databases allows the engineer to select a material, and those material properties are automatically called into the relevant analysis code. The ADP2 materials database will be composed of four independent schemas: CAE Design, Processing, Testing, and Logistics Support. The design of ADP2 places major emphasis on the seamless integration of CAE and analysis modules with a single intuitive graphical interface. This tool is being designed to serve and be used by an entire project team, i.e., analysts, designers, materials experts, and managers. The final version of the software will be delivered to the Air Force in Jan. 1994. The Analytical Design Package (ADP2) will then be ready for transfer to industry. The package will be capable of a wide range of design and manufacturing applications.

  15. File list: InP.Adp.05.AllAg.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.05.AllAg.AllCell mm9 Input control Adipocyte SRX997757,SRX478163,SRX821800,...0,SRX341056,SRX341058,SRX821799,SRX341057,SRX341783,SRX341781,SRX478161,SRX127367,SRX127370 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.05.AllAg.AllCell.bed ...

  16. File list: InP.Adp.50.AllAg.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.AllAg.AllCell mm9 Input control Adipocyte SRX185879,SRX143805,SRX268023,...7,SRX341056,SRX341058,SRX821800,SRX821801,SRX821799,SRX478163,SRX478161,SRX127370,SRX127367 http://dbarchive.biosciencedbc.jp/kyushu-u/mm9/assembled/InP.Adp.50.AllAg.AllCell.bed ...

  17. Pistacia chinensis Methanolic Extract Attenuated MAPK and Akt Phosphorylations in ADP Stimulated Rat Platelets In Vitro

    OpenAIRE

    Ji Young Park; Mei Hong; Qi Jia; Young-Chul Lee; Taddesse Yayeh; Eujin Hyun; Dong-Mi Kwak; Jae Youl Cho; Man Hee Rhee

    2012-01-01

    Pistacia chinensis (Chinese pistache) is a widely grown plant in southern China where the galls extract is a common practice in folk medicine. However, extracts from this plant have never been attempted for their cardiovascular protective effects in experimental setting. Here therefore we aimed to investigate the antiplatelet activity of Pistacia chinensis methanolic extract (PCME) in ADP stimulated rat platelets in vitro. PCME (2.5–20 μg/mL) inhibited ADP-induced platelet aggregation. While ...

  18. Pistacia chinensis Methanolic Extract Attenuated MAPK and Akt Phosphorylations in ADP Stimulated Rat Platelets In Vitro

    Directory of Open Access Journals (Sweden)

    Ji Young Park

    2012-01-01

    (2.5–20 μg/mL inhibited ADP-induced platelet aggregation. While PCME diminished [Ca2+]i, ATP, and TXA2 release in ADP-activated platelets, it enhanced cAMP production in resting platelets. Likewise, PCME inhibited fibrinogen binding to αIIbβ3 and downregulated JNK, ERK, and Akt phosphorylations. Thus, PCME contains potential antiplatelet compounds that could be deployed for their therapeutic values in cardiovascular pathology.

  19. Two Arabidopsis ADP-glucose pyrophosphorylase large subunits (APL1 and APL2) are catalytic.

    Science.gov (United States)

    Ventriglia, Tiziana; Kuhn, Misty L; Ruiz, Ma Teresa; Ribeiro-Pedro, Marina; Valverde, Federico; Ballicora, Miguel A; Preiss, Jack; Romero, José M

    2008-09-01

    ADP-glucose (Glc) pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in starch biosynthesis. Higher plant ADP-Glc PPase is a heterotetramer (alpha(2)beta(2)) consisting of two small and two large subunits. There is increasing evidence that suggests that catalytic and regulatory properties of the enzyme from higher plants result from the synergy of both types of subunits. In Arabidopsis (Arabidopsis thaliana), two genes encode small subunits (APS1 and APS2) and four large subunits (APL1-APL4). Here, we show that in Arabidopsis, APL1 and APL2, besides their regulatory role, have catalytic activity. Heterotetramers formed by combinations of a noncatalytic APS1 and the four large subunits showed that APL1 and APL2 exhibited ADP-Glc PPase activity with distinctive sensitivities to the allosteric activator (3-phosphoglycerate). Mutation of the Glc-1-P binding site of Arabidopsis and potato (Solanum tuberosum) isoforms confirmed these observations. To determine the relevance of these activities in planta, a T-DNA mutant of APS1 (aps1) was characterized. aps1 is starchless, lacks ADP-Glc PPase activity, APS1 mRNA, and APS1 protein, and is late flowering in long days. Transgenic lines of the aps1 mutant, expressing an inactivated form of APS1, recovered the wild-type phenotype, indicating that APL1 and APL2 have catalytic activity and may contribute to ADP-Glc synthesis in planta. PMID:18614708

  20. Two Arabidopsis ADP-Glucose Pyrophosphorylase Large Subunits (APL1 and APL2) Are Catalytic1

    Science.gov (United States)

    Ventriglia, Tiziana; Kuhn, Misty L.; Ruiz, Ma Teresa; Ribeiro-Pedro, Marina; Valverde, Federico; Ballicora, Miguel A.; Preiss, Jack; Romero, José M.

    2008-01-01

    ADP-glucose (Glc) pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step in starch biosynthesis. Higher plant ADP-Glc PPase is a heterotetramer (α2β2) consisting of two small and two large subunits. There is increasing evidence that suggests that catalytic and regulatory properties of the enzyme from higher plants result from the synergy of both types of subunits. In Arabidopsis (Arabidopsis thaliana), two genes encode small subunits (APS1 and APS2) and four large subunits (APL1–APL4). Here, we show that in Arabidopsis, APL1 and APL2, besides their regulatory role, have catalytic activity. Heterotetramers formed by combinations of a noncatalytic APS1 and the four large subunits showed that APL1 and APL2 exhibited ADP-Glc PPase activity with distinctive sensitivities to the allosteric activator (3-phosphoglycerate). Mutation of the Glc-1-P binding site of Arabidopsis and potato (Solanum tuberosum) isoforms confirmed these observations. To determine the relevance of these activities in planta, a T-DNA mutant of APS1 (aps1) was characterized. aps1 is starchless, lacks ADP-Glc PPase activity, APS1 mRNA, and APS1 protein, and is late flowering in long days. Transgenic lines of the aps1 mutant, expressing an inactivated form of APS1, recovered the wild-type phenotype, indicating that APL1 and APL2 have catalytic activity and may contribute to ADP-Glc synthesis in planta. PMID:18614708

  1. Bacterial carbonatogenesis

    International Nuclear Information System (INIS)

    Several series of experiments in the laboratory as well as in natural conditions teach that the production of carbonate particles by heterotrophic bacteria follows different ways. The 'passive' carbonatogenesis is generated by modifications of the medium that lead to the accumulation of carbonate and bicarbonate ions and to the precipitation of solid particles. The 'active' carbonatogenesis is independent of the metabolic pathways. The carbonate particles are produced by ionic exchanges through the cell membrane following still poorly known mechanisms. Carbonatogenesis appears to be the response of heterotrophic bacterial communities to an enrichment of the milieu in organic matter. The active carbonatogenesis seems to start first. It is followed by the passive one which induces the growth of initially produced particles. The yield of heterotrophic bacterial carbonatogenesis and the amounts of solid carbonates production by bacteria are potentially very high as compared to autotrophic or chemical sedimentation from marine, paralic or continental waters. Furthermore, the bacterial processes are environmentally very ubiquitous; they just require organic matter enrichment. Thus, apart from purely evaporite and autotrophic ones, all Ca and/or Mg carbonates must be considered as from heterotrophic bacterial origin. By the way, the carbon of carbonates comes from primary organic matter. Such considerations ask questions about some interpretations from isotopic data on carbonates. Finally, bacterial heterotrophic carbonatogenesis appears as a fundamental phase in the relationships between atmosphere and lithosphere and in the geo-biological evolution of Earth. (author)

  2. Bacterial Adhesion & Blocking Bacterial Adhesion

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk

    2008-01-01

    tract to the microbial flocs in waste water treatment facilities. Microbial biofilms may however also cause a wide range of industrial and medical problems, and have been implicated in a wide range of persistent infectious diseases, including implantassociated microbial infections. Bacterial adhesion...... is the first committing step in biofilm formation, and has therefore been intensely scrutinized. Much however, still remains elusive. Bacterial adhesion is a highly complex process, which is influenced by a variety of factors. In this thesis, a range of physico-chemical, molecular and environmental parameters......, which influence the transition from a planktonic lifestyle to a sessile lifestyle, have been studied. Protein conditioning film formation was found to influence bacterial adhesion and subsequent biofilm formation considerable, and an aqueous extract of fish muscle tissue was shown to significantly...

  3. Bacterial lipases

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

    1994-01-01

    Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation, mea

  4. Accurate determination of the oxidative phosphorylation affinity for ADP in isolated mitochondria.

    Directory of Open Access Journals (Sweden)

    Gilles Gouspillou

    Full Text Available BACKGROUND: Mitochondrial dysfunctions appear strongly implicated in a wide range of pathologies. Therefore, there is a growing need in the determination of the normal and pathological integrated response of oxidative phosphorylation to cellular ATP demand. The present study intends to address this issue by providing a method to investigate mitochondrial oxidative phosphorylation affinity for ADP in isolated mitochondria. METHODOLOGY/PRINCIPAL FINDINGS: The proposed method is based on the simultaneous monitoring of substrate oxidation (determined polarographically and phosphorylation (determined using the glucose-hexokinase glucose-6-phosphate dehydrogenase-NADP(+ enzymatic system rates, coupled to the determination of actual ADP and ATP concentrations by bioluminescent assay. This enzymatic system allows the study of oxidative phosphorylation during true steady states in a wide range of ADP concentrations. We demonstrate how the application of this method allows an accurate determination of mitochondrial affinity for ADP from both oxidation (K(mVox and phosphorylation (K(mVp rates. We also demonstrate that determination of K(mVox leads to an important overestimation of the mitochondrial affinity for ADP, indicating that mitochondrial affinity for ADP should be determined using phosphorylation rate. Finally, we show how this method allows the direct and precise determination of the mitochondrial coupling efficiency. Data obtained from rat skeletal muscle and liver mitochondria illustrate the discriminating capabilities of this method. CONCLUSIONS/SIGNIFICANCE: Because the proposed method allows the accurate determination of mitochondrial oxidative phosphorylation affinity for ADP in isolated mitochondria, it also opens the route to a better understanding of functional consequences of mitochondrial adaptations/dysfunctions arising in various physiological/pathophysiological conditions.

  5. Adenovirus Ad-p53AIP1-mediated gene therapy and its regulation of p53-MDM2 interactions

    OpenAIRE

    Jiang, Yunbo; Chen, Huihua; JIA, HAIQUAN; XU, YUANJI; Liu, Gang; Wang, Yan; Yang, Xiaohe; Yinglin LU

    2010-01-01

    We generated replication-defective adenovirus Ad-p53AIP1 and studied its anti-tumor efficacy both in vitro and in vivo. We demonstrated that Ad-p53AIP1 infection elicited high levels of p53AIP1 expression in cancer cells. We also found that Ad-p53AIP1 expression induced marked apoptosis and cell cycle arrest in HepG2 cells. Moreover, Ad-p53AIP1 infection significantly inhibited the tumorigenesis of 4T1 mouse mammary cancer cells in vivo. In particular, we discovered that p53AIP1 overexpressio...

  6. Synergistic role of ADP and Ca(2+) in diastolic myocardial stiffness.

    Science.gov (United States)

    Sequeira, Vasco; Najafi, Aref; McConnell, Mark; Fowler, Ewan D; Bollen, Ilse A E; Wüst, Rob C I; dos Remedios, Cris; Helmes, Michiel; White, Ed; Stienen, Ger J M; Tardiff, Jil; Kuster, Diederik W D; van der Velden, Jolanda

    2015-09-01

    Diastolic dysfunction in heart failure patients is evident from stiffening of the passive properties of the ventricular wall. Increased actomyosin interactions may significantly limit diastolic capacity, however, direct evidence is absent. From experiments at the cellular and whole organ level, in humans and rats, we show that actomyosin-related force development contributes significantly to high diastolic stiffness in environments where high ADP and increased diastolic [Ca(2+) ] are present, such as the failing myocardium. Our basal study provides a mechanical mechanism which may partly underlie diastolic dysfunction. Heart failure (HF) with diastolic dysfunction has been attributed to increased myocardial stiffness that limits proper filling of the ventricle. Altered cross-bridge interaction may significantly contribute to high diastolic stiffness, but this has not been shown thus far. Cross-bridge interactions are dependent on cytosolic [Ca(2+) ] and the regeneration of ATP from ADP. Depletion of myocardial energy reserve is a hallmark of HF leading to ADP accumulation and disturbed Ca(2+) handling. Here, we investigated if ADP elevation in concert with increased diastolic [Ca(2+) ] promotes diastolic cross-bridge formation and force generation and thereby increases diastolic stiffness. ADP dose-dependently increased force production in the absence of Ca(2+) in membrane-permeabilized cardiomyocytes from human hearts. Moreover, physiological levels of ADP increased actomyosin force generation in the presence of Ca(2+) both in human and rat membrane-permeabilized cardiomyocytes. Diastolic stress measured at physiological lattice spacing and 37°C in the presence of pathological levels of ADP and diastolic [Ca(2+) ] revealed a 76 ± 1% contribution of cross-bridge interaction to total diastolic stress in rat membrane-permeabilized cardiomyocytes. Inhibition of creatine kinase (CK), which increases cytosolic ADP, in enzyme-isolated intact rat cardiomyocytes impaired

  7. Selective down-regulation of nuclear poly(ADP-ribose glycohydrolase.

    Directory of Open Access Journals (Sweden)

    David M Burns

    Full Text Available BACKGROUND: The formation of ADP-ribose polymers on target proteins by poly(ADP-ribose polymerases serves a variety of cell signaling functions. In addition, extensive activation of poly(ADP-ribose polymerase-1 (PARP-1 is a dominant cause of cell death in ischemia-reperfusion, trauma, and other conditions. Poly(ADP-ribose glycohydrolase (PARG degrades the ADP-ribose polymers formed on acceptor proteins by PARP-1 and other PARP family members. PARG exists as multiple isoforms with differing subcellular localizations, but the functional significance of these isoforms is uncertain. METHODS / PRINCIPAL FINDINGS: Primary mouse astrocytes were treated with an antisense phosphorodiamidate morpholino oligonucleotide (PMO targeted to exon 1 of full-length PARG to suppress expression of this nuclear-specific PARG isoform. The antisense-treated cells showed down-regulation of both nuclear PARG immunoreactivity and nuclear PARG enzymatic activity, without significant alteration in cytoplasmic PARG activity. When treated with the genotoxic agent MNNG to induced PARP-1 activation, the antisense-treated cells showed a delayed rate of nuclear PAR degradation, reduced nuclear condensation, and reduced cell death. CONCLUSIONS/SIGNIFICANCE: These results support a preferentially nuclear localization for full-length PARG, and suggest a key role for this isoform in the PARP-1 cell death pathway.

  8. Study on A.C. electrical properties of pure and L-serine doped ADP crystals

    Science.gov (United States)

    Joshi, J. H.; Dixit, K. P.; Joshi, M. J.; Parikh, K. D.

    2016-05-01

    Ammonium Dihydrogen Phosphate (ADP) crystals have a wide range of applications in integrated and nonlinear optics. Amino acids having significant properties like molecular chirality, zwitter ionic nature, etc. attracted many researchers to dope them in various NLO crystals. In the present study, pure and different weight percentage L-serine doped ADP crystals were grown by slow solvent evaporation technique at room temperature. The A.C. electrical study was carried out for palletized samples at room temperature. The Nyquist plot showed two semi circles for pure ADP indicated the effect of grain and grain boundary, whereas the doped ADP samples exhibited the single semi circle suggesting the effect of grain. The values resistance and capacitance for grain and grain boundary were calculated. The effect of doping was clearly seen in the grain capacitance and resistance values. The dielectric constant and dielectric loss decreased with increase in frequency for all samples. The Jonscher power law was applied for A.C. conductivity for pure and doped ADP samples. The imaginary part of modulus and impedance versus frequency were drawn and the value of stretch exponent (β) was calculated for all the samples.

  9. A fluorometric assay for measurement of mono-ADP-ribosyltransferase activity.

    Science.gov (United States)

    Klebl, B M; Pette, D

    1996-08-01

    Using 1,N6-etheno NAD, a fluorescent analog of NAD, we extended an existing assay for NAD glycohydrolase to the measurement of mono-ADP-ribosyltransferase (mADP-RT) activity using agmatine as acceptor for ADP-ribose. The reaction products were analyzed by reversed-phase chromatography. In the presence of agmatine two newly formed fluorescent products were tentatively identified as ADP-ribosylagmatine anomers. Fluorescence intensity increased upon splitting the N-glycoside bondage of 1,N6-etheno NAD. Therefore, 1, N6-etheno AMP could be used for calibration. The nonradioactive assay yielded values nearly identical to those obtained with the [carbonyl-14C]NAD method. It proved to be highly reproducible, rapid, and suitable for an improved purification protocol yielding a 76,000-fold enriched mADP-RT preparation from rabbit skeletal muscle. The identity and high purity of the enzyme were confirmed immunochemically. The assay served to determine the pH optimum of the enzyme (pH 9.0) and its KM for 1,N6-etheno NAD (287 microM). PMID:8811894

  10. Bacterial Ecology

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2011-01-01

    Bacterial ecology is concerned with the interactions between bacteria and their biological and nonbiological environments and with the role of bacteria in biogeochemical element cycling. Many fundamental properties of bacteria are consequences of their small size. Thus, they can efficiently exploit...... biogeochemical processes are carried exclusively by bacteria. * Bacteria play an important role in all types of habitats including some that cannot support eukaryotic life....

  11. [Bacterial vaginosis].

    Science.gov (United States)

    Romero Herrero, Daniel; Andreu Domingo, Antonia

    2016-07-01

    Bacterial vaginosis (BV) is the main cause of vaginal dysbacteriosis in the women during the reproductive age. It is an entity in which many studies have focused for years and which is still open for discussion topics. This is due to the diversity of microorganisms that cause it and therefore, its difficult treatment. Bacterial vaginosis is probably the result of vaginal colonization by complex bacterial communities, many of them non-cultivable and with interdependent metabolism where anaerobic populations most likely play an important role in its pathogenesis. The main symptoms are an increase of vaginal discharge and the unpleasant smell of it. It can lead to serious consequences for women, such as an increased risk of contracting sexually transmitted infections including human immunodeficiency virus and upper genital tract and pregnancy complications. Gram stain is the gold standard for microbiological diagnosis of BV, but can also be diagnosed using the Amsel clinical criteria. It should not be considered a sexually transmitted disease but it is highly related to sex. Recurrence is the main problem of medical treatment. Apart from BV, there are other dysbacteriosis less characterized like aerobic vaginitis of which further studies are coming slowly but are achieving more attention and consensus among specialists. PMID:27474242

  12. Effect of Co2+ doping on solubility, crystal growth and properties of ADP crystals

    Science.gov (United States)

    Ganesh, V.; Shkir, Mohd.; AlFaify, S.; Yahia, I. S.

    2016-09-01

    Bulk size crystal growth of ADP with different concentrations doping of cobalt (Co2+) has been done by low cost slow evaporation technique at ambient conditions. The solubility measurement was carried out on pure and doped crystals and found that the solubility is decreasing with doping concentrations. The presence of Co2+ ion in crystalline matrix of ADP has been confirmed by structural, vibrational and elemental analyses. Scanning electron microscopic study reveals that the doping has strong effect on the quality of the crystals. The optical absorbance and transmission confirms the enhancement of quality of ADP crystals due to Co2+ doping and so the optical band gap. Further the dislocation, photoluminescence, dielectric and mechanical studies confirms that the properties of grown crystals with Co2+ doping has been enriched and propose it as a better candidate for optoelectronic applications.

  13. Pistacia chinensis Methanolic Extract Attenuated MAPK and Akt Phosphorylations in ADP Stimulated Rat Platelets In Vitro.

    Science.gov (United States)

    Park, Ji Young; Hong, Mei; Jia, Qi; Lee, Young-Chul; Yayeh, Taddesse; Hyun, Eujin; Kwak, Dong-Mi; Cho, Jae Youl; Rhee, Man Hee

    2012-01-01

    Pistacia chinensis (Chinese pistache) is a widely grown plant in southern China where the galls extract is a common practice in folk medicine. However, extracts from this plant have never been attempted for their cardiovascular protective effects in experimental setting. Here therefore we aimed to investigate the antiplatelet activity of Pistacia chinensis methanolic extract (PCME) in ADP stimulated rat platelets in vitro. PCME (2.5-20 μg/mL) inhibited ADP-induced platelet aggregation. While PCME diminished [Ca(2+)]i, ATP, and TXA2 release in ADP-activated platelets, it enhanced cAMP production in resting platelets. Likewise, PCME inhibited fibrinogen binding to αIIbβ3 and downregulated JNK, ERK, and Akt phosphorylations. Thus, PCME contains potential antiplatelet compounds that could be deployed for their therapeutic values in cardiovascular pathology. PMID:22899962

  14. Multiple origins of hydrogenosomes : functional and phylogenetic evidence from the ADP/ATP carrier of the anaerobic chytrid Neocallimastix sp.

    NARCIS (Netherlands)

    Voncken, F; Boxma, B; Tjaden, J; Akhmanova, A; Huynen, M; Tielens, AGM; Haferkamp, [No Value; Neuhaus, HE; Vogels, G; Veenhuis, M; Hackstein, JHP; Tielens, Aloysius G.M.; Haferkamp, Ilka; Hackstein, Johannes H.P.

    2002-01-01

    A mitochondrial-type ADP/ATP carrier (AAC) has been identified in the hydrogenosomes of the anaerobic chytridiomycete fungus Neocallimastix sp. L2. Biochemical and immunocytochemical studies revealed that this ADP/ATP carrier is an integral component of hydrogenosomal membranes. Expression of the co

  15. File list: His.Adp.05.H2BS112GlcNAc.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  16. File list: His.Adp.20.H2BS112GlcNAc.AllCell [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  17. File list: InP.Adp.50.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  18. Higher cytoplasmic and nuclear poly(ADP-ribose) polymerase expression in familial than in sporadic breast cancer

    NARCIS (Netherlands)

    Klauke, M.L.; Hoogerbrugge-van der Linden, N.; Budczies, J.; Bult, P.; Prinzler, J.; Radke, C.; Krieken, J.H. van; Dietel, M.; Denkert, C.; Muller, B.M.

    2012-01-01

    Poly(ADP-ribose) polymerase 1 (PARP) is a key element of the single-base excision pathway for repair of DNA single-strand breaks. To compare the cytoplasmic and nuclear poly(ADP-ribose) expression between familial (BRCA1, BRCA2, or non BRCA1/2) and sporadic breast cancer, we investigated 39 sporadic

  19. File list: InP.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.20.AllAg.Adipose_Tissue,_White hg19 Input control Adipocyte Adipose Tissue,... White http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.20.AllAg.Adipose_Tissue,_White.bed ...

  20. File list: NoD.Adp.20.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

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  6. File list: InP.Adp.20.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

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  7. File list: InP.Adp.50.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

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  9. File list: InP.Adp.10.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

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  11. File list: InP.Adp.05.AllAg.Adipose_progenitor_cells [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  12. File list: NoD.Adp.10.AllAg.Adipose_stromal_cell [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available NoD.Adp.10.AllAg.Adipose_stromal_cell hg19 No description Adipocyte Adipose stromal... cell http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/NoD.Adp.10.AllAg.Adipose_stromal_cell.bed ...

  13. Molecular and biochemical analysis of the plastidic ADP-glucose transporter (ZmBT1) from Zea mays

    NARCIS (Netherlands)

    Kirchberger, S.; Leroch, M.; Huynen, M.A.; Wahl, M.; Neuhaus, H.E.; Tjaden, J.

    2007-01-01

    Physiological studies on the Brittle1 maize mutant have provided circumstantial evidence that ZmBT1 (Zea mays Brittle1 protein) is involved in the ADP-Glc transport into maize endosperm plastids, but up to now, no direct ADP-Glc transport mediated by ZmBT1 has ever been shown. The heterologous synth

  14. Molecular and biochemical analysis of the plastidic ADP-glucose transporter (ZmBT1) from Zea mays.

    NARCIS (Netherlands)

    Kirchberger, S.; Leroch, M.; Huynen, M.A.; Wahl, M.; Neuhaus, H.E.; Tjaden, J.

    2007-01-01

    Physiological studies on the Brittle1 maize mutant have provided circumstantial evidence that ZmBT1 (Zea mays Brittle1 protein) is involved in the ADP-Glc transport into maize endosperm plastids, but up to now, no direct ADP-Glc transport mediated by ZmBT1 has ever been shown. The heterologous synth

  15. File list: InP.Adp.50.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

    Full Text Available InP.Adp.50.AllAg.Adipose_Tissue,_White hg19 Input control Adipocyte Adipose Tissue, White... http://dbarchive.biosciencedbc.jp/kyushu-u/hg19/assembled/InP.Adp.50.AllAg.Adipose_Tissue,_White.bed ...

  16. File list: InP.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

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    Lifescience Database Archive (English)

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  18. File list: NoD.Adp.10.AllAg.Adipose_Tissue,_White [Chip-atlas[Archive

    Lifescience Database Archive (English)

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  19. Correlations of serum levels of leptin and other related factor (NPY, ADP) in female children with simple obesity

    International Nuclear Information System (INIS)

    Objective: To study the changes of serum levels of leptin, NPY and ADP in female children with simple obesity. Methods: Serum levels of leptin, NPY and ADP were measured with radioimmunoassay (RIA) in 32 female children with simple obesity and 35 controls. Results: The serum levels of leptin, NPY were significantly higher in the obese children than those in controls (P<0.01), while the serum levels of ADP were significantly lower (P<0.01). Serum leptin levels were significantly positively correlated (r=0.6014, P<0.01) with NPY levels but were negatively correlated (r=-0.4786, P<0.01) with adiponectin (ADP) levels. Conclusion: Determination of serum leptin, NPY and ADP levels is of help for judgement of degree of obesity as wen as outcome prediction in female children. (authors)

  20. ADP-bildende Acetyl-CoA Synthetasen aus hyperthermophilen Archaea: Molekularbiologische und biochemische Charakterisierung von neuartigen Enzymen der Acetat-Bildung und ATP-Synthese

    OpenAIRE

    Musfeldt, Meike

    2001-01-01

    Keine deutschsprachige Zusammenfassung vorhanden. Acetyl-CoA synthetase (ADP-forming) (ADP-ACS) represents a novel enzyme of acetate formation and energy conservation (acetyl-CoA + ADP + Pi -> acetate + ATP + CoA) in Archaea and eukaryotic protists. The only characterized ADP-ACS in Archaea, two isoenzymes from the hyperthermophile Pyrococcus furiosus, constitute 145 kDa heterotetramers (a2, b2). By using the N-terminal amino acid sequences of both subunits, which are located at different ...

  1. Improved Triacylglycerol Production in Acinetobacter baylyi ADP1 by Metabolic Engineering

    Directory of Open Access Journals (Sweden)

    Karp Matti

    2011-05-01

    Full Text Available Abstract Background Triacylglycerols are used in various purposes including food applications, cosmetics, oleochemicals and biofuels. Currently the main sources for triacylglycerol are vegetable oils, and microbial triacylglycerol has been suggested as an alternative for these. Due to the low production rates and yields of microbial processes, the role of metabolic engineering has become more significant. As a robust model organism for genetic and metabolic studies, and for the natural capability to produce triacylglycerol, Acinetobacter baylyi ADP1 serves as an excellent organism for modelling the effects of metabolic engineering for energy molecule biosynthesis. Results Beneficial gene deletions regarding triacylglycerol production were screened by computational means exploiting the metabolic model of ADP1. Four deletions, acr1, poxB, dgkA, and a triacylglycerol lipase were chosen to be studied experimentally both separately and concurrently by constructing a knock-out strain (MT with three of the deletions. Improvements in triacylglycerol production were observed: the strain MT produced 5.6 fold more triacylglycerol (mg/g cell dry weight compared to the wild type strain, and the proportion of triacylglycerol in total lipids was increased by 8-fold. Conclusions In silico predictions of beneficial gene deletions were verified experimentally. The chosen single and multiple gene deletions affected beneficially the natural triacylglycerol metabolism of A. baylyi ADP1. This study demonstrates the importance of single gene deletions in triacylglycerol metabolism, and proposes Acinetobacter sp. ADP1 as a model system for bioenergetic studies regarding metabolic engineering.

  2. Aero-Propulsion Technology (APT) Task V Low Noise ADP Engine Definition Study

    Science.gov (United States)

    Holcombe, V.

    2003-01-01

    A study was conducted to identify and evaluate noise reduction technologies for advanced ducted prop propulsion systems that would allow increased capacity operation and result in an economically competitive commercial transport. The study investigated the aero/acoustic/structural advancements in fan and nacelle technology required to match or exceed the fuel burned and economic benefits of a constrained diameter large Advanced Ducted Propeller (ADP) compared to an unconstrained ADP propulsion system with a noise goal of 5 to 10 EPNDB reduction relative to FAR 36 Stage 3 at each of the three measuring stations namely, takeoff (cutback), approach and sideline. A second generation ADP was selected to operate within the maximum nacelle diameter constrain of 160 deg to allow installation under the wing. The impact of fan and nacelle technologies of the second generation ADP on fuel burn and direct operating costs for a typical 3000 nm mission was evaluated through use of a large, twin engine commercial airplane simulation model. The major emphasis of this study focused on fan blade aero/acoustic and structural technology evaluations and advanced nacelle designs. Results of this study have identified the testing required to verify the interactive performance of these components, along with noise characteristics, by wind tunnel testing utilizing and advanced interaction rig.

  3. 32 CFR Appendix J to Part 154 - ADP Position Categories and Criteria for Designating Positions

    Science.gov (United States)

    2010-07-01

    ... Designating Positions J Appendix J to Part 154 National Defense Department of Defense OFFICE OF THE SECRETARY OF DEFENSE SECURITY DEPARTMENT OF DEFENSE PERSONNEL SECURITY PROGRAM REGULATION Pt. 154, App. J Appendix J to Part 154—ADP Position Categories and Criteria for Designating Positions OMB Circular...

  4. Impaired ADP channeling to mitochondria and elevated reactive oxygen species in hypertensive hearts.

    Science.gov (United States)

    Power, Amelia S C; Pham, Toan; Loiselle, Denis S; Crossman, David H; Ward, Marie-Louise; Hickey, Anthony J

    2016-06-01

    Systemic hypertension initially promotes a compensatory cardiac hypertrophy, yet it progresses to heart failure (HF), and energetic deficits appear to be central to this failure. However, the transfer of energy between the mitochondria and the myofibrils is not often considered as part of the energetic equation. We compared hearts from old spontaneously hypertensive rats (SHRs) and normotensive Wistar controls. SHR hearts showed a 35% depression in mitochondrial function, yet produced at least double the amount of reactive oxygen species (ROS) in all respiration states in left ventricular (LV) homogenates. To test the connectivity between mitochondria and myofibrils, respiration was further tested in situ with LV permeabilized fibers by addition of multiple substrates and ATP, which requires hydrolysis to mediate oxidative phosphorylation. By trapping ADP using a pyruvate kinase enzyme system, we tested ADP channeling towards mitochondria, and this suppressed respiration and elevated ROS production more in the SHR fibers. The ADP-trapped state was also less relieved on creatine addition, likely reflecting the 30% depression in total CK activity in the SHR heart fibers. Confocal imaging identified a 34% longer distance between the centers of myofibril to mitochondria in the SHR hearts, which increases transverse metabolite diffusion distances (e.g., for ATP, ADP, and creatine phosphate). We propose that impaired connectivity between mitochondria and myofibrils may contribute to elevated ROS production. Impaired energy exchange could be the result of ultrastructural changes that occur with hypertrophy in this model of hypertension. PMID:27084386

  5. Skeletal muscle contractile performance and ADP accumulation in adenylate kinase-deficient mice

    NARCIS (Netherlands)

    Hancock, C.R.; Janssen, E.E.W.; Terjung, R.L.

    2005-01-01

    The production of AMP by adenylate kinase (AK) and subsequent deamination by AMP deaminase limits ADP accumulation during conditions of high-energy demand in skeletal muscle. The goal of this study was to investigate the consequences of AK deficiency (-/-) on adenine nucleotide management and whole

  6. 45 CFR 95.625 - Increased FFP for certain ADP systems.

    Science.gov (United States)

    2010-10-01

    ...-D program are contained in 45 CFR Part 307. The applicable regulations for the Title IV-E program are contained in 45 CFR 1355.55. The applicable regulations for the Title XIX program are contained in 42 CFR Part 433, Subpart C. Federal Financial Participation in Costs of ADP Acquisitions...

  7. Bacterial Hydrodynamics

    Science.gov (United States)

    Lauga, Eric

    2016-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells, yet they represent the bulk of the world's biomass and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micrometer scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically complex environments. Using hydrodynamics as an organizing framework, I review the biomechanics of bacterial motility and look ahead to future challenges.

  8. Bacterial hydrodynamics

    CERN Document Server

    Lauga, Eric

    2015-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells yet they represent the bulk of the world's biomass, and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds-number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micron scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically-complex environments. Using hydrodynamics as an organizing framework, we review the biomechanics of bacterial motility and look ahead to future challenges.

  9. The ancestral activation promiscuity of ADP-glucose pyrophosphorylases from oxygenic photosynthetic organisms

    Directory of Open Access Journals (Sweden)

    Kuhn Misty L

    2013-02-01

    Full Text Available Abstract Background ADP-glucose pyrophosphorylase (ADP-Glc PPase catalyzes the first committed step in the synthesis of glycogen in bacteria and starch in algae and plants. In oxygenic photosynthetic organisms, ADP-Glc PPase is mainly activated by 3-phosphoglycerate (3-PGA and to a lesser extent by other metabolites. In this work, we analyzed the activation promiscuity of ADP-Glc PPase subunits from the cyanobacterium Anabaena PCC 7120, the green alga Ostreococcus tauri, and potato (Solanum tuberosum tuber by comparing a specificity constant for 3-PGA, fructose-1,6-bisphosphate (FBP, fructose-6-phosphate, and glucose-6-phosphate. Results The 3-PGA specificity constant for the enzymes from Anabaena (homotetramer, O. tauri, and potato tuber was considerably higher than for other activators. O. tauri and potato tuber enzymes were heterotetramers comprising homologous small and large subunits. Conversely, the O. tauri small subunit (OtaS homotetramer was more promiscuous because its FBP specificity constant was similar to that for 3-PGA. To explore the role of both OtaS and OtaL (O. tauri large subunit in determining the specificity of the heterotetramer, we knocked out the catalytic activity of each subunit individually by site-directed mutagenesis. Interestingly, the mutants OtaSD148A/OtaL and OtaS/OtaLD171A had higher specificity constants for 3-PGA than for FBP. Conclusions After gene duplication, OtaS seemed to have lost specificity for 3-PGA compared to FBP. This was physiologically and evolutionarily feasible because co-expression of both subunits restored the specificity for 3-PGA of the resulting heterotetrameric wild type enzyme. This widespread promiscuity seems to be ancestral and intrinsic to the enzyme family. Its presence could constitute an efficient evolutionary mechanism to accommodate the ADP-Glc PPase regulation to different metabolic needs.

  10. Adenosine diphosphate ribosylation of dinitrogenase reductase and adenylylation of glutamine synthetase control ammonia excretion in ethylenediamine-resistant mutants of Azospirillum brasilense Sp7.

    Science.gov (United States)

    Srivastava, A; Tripathi, A K

    2006-10-01

    Azospirillum brasilense is a nitrogen-fixing, root-colonizing bacterium that brings about plant-growth-promoting effects mainly because of its ability to produce phytohormones. Ethylenediamine (EDA)-resistant mutants of A. brasilense were isolated and screened for their higher ability to decrease acetylene and release ammonia in the medium. One of the mutants showed considerably higher levels of acetylene decrease and ammonia excretion. Nitrogenase activity of this mutant was relatively resistant to inhibition by NH(4)Cl. Adenosine triphosphate ribosylation of dinitrogenase reductase in the mutant did not increase even in presence of 10 mM NH(4)Cl. Although the mutant showed decreased glutamine synthetase (GS) activity, neither the levels of GS synthesized by the mutant nor the NH (4) (+) -binding site in the GS differed from those of the parent. The main reason for the release of ammonia by the mutant seems to be the fixation of higher levels of nitrogen than its GS can assimilate, as well as higher levels of adenylylation of GS, which may decrease ammonia assimilation.

  11. 阿维链霉菌adpA-a调控形态分化和黑色素形成

    Institute of Scientific and Technical Information of China (English)

    赵金雷; 文莹; 陈芝; 宋渊; 李季伦

    2007-01-01

    灰色链霉菌(Streptomyces griseus)中的AdpA是A-因子调控网络中的一个中心转录调控因子,控制形态分化和次级代谢.在阿维链霉菌(Streptomyces avermitilis)的基因组上,也存在着与adpA高度同源的基因adpA-a.为了研究其功能,通过同源双交换将adpA-a基因破坏,得到的adpA-a突变株不能进行正常的形态分化,同时不再产生黑色素,但产阿维菌素(avermectin)的能力不受影响.对该破坏菌株进行基因互补,互补突变株的表型得到恢复,证实了突变株表型变化是由adpA-a破坏引起的.以上结果表明,在阿维链霉菌中,adpA-a不仅参与形态分化的调控,同时也参与黑色素生物合成的调控.

  12. Altered poly(ADP-ribose) metabolism impairs cellular responses to genotoxic stress in a hypomorphic mutant of poly(ADP-ribose) glycohydrolase

    International Nuclear Information System (INIS)

    Genotoxic stress activates nuclear poly(ADP-ribose) (PAR) metabolism leading to PAR synthesis catalyzed by DNA damage activated poly(ADP-ribose) polymerases (PARPs) and rapid PAR turnover by action of nuclear poly(ADP-ribose) glycohydrolase (PARG). The involvement of PARP-1 and PARP-2 in responses to DNA damage has been well studied but the involvement of nuclear PARG is less well understood. To gain insights into the function of nuclear PARG in DNA damage responses, we have quantitatively studied PAR metabolism in cells derived from a hypomorphic mutant mouse model in which exons 2 and 3 of the PARG gene have been deleted (PARG-Δ2,3 cells), resulting in a nuclear PARG containing a catalytic domain but lacking the N-terminal region (A domain) of the protein. Following DNA damage induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), we found that the activity of both PARG and PARPs in intact cells is increased in PARG-Δ2,3 cells. The increased PARG activity leads to decreased PARP-1 automodification with resulting increased PARP activity. The degree of PARG activation is greater than PARP, resulting in decreased PAR accumulation. Following MNNG treatment, PARG-Δ2,3 cells show reduced formation of XRCC1 foci, delayed H2AX phosphorylation, decreased DNA break intermediates during repair, and increased cell death. Our results show that a precise coordination of PARPs and PARG activities is important for normal cellular responses to DNA damage and that this coordination is defective in the absence of the PARG A domain

  13. Semicarbazone EGA Inhibits Uptake of Diphtheria Toxin into Human Cells and Protects Cells from Intoxication

    Directory of Open Access Journals (Sweden)

    Leonie Schnell

    2016-07-01

    Full Text Available Diphtheria toxin is a single-chain protein toxin that invades human cells by receptor-mediated endocytosis. In acidic endosomes, its translocation domain inserts into endosomal membranes and facilitates the transport of the catalytic domain (DTA from endosomal lumen into the host cell cytosol. Here, DTA ADP-ribosylates elongation factor 2 inhibits protein synthesis and leads to cell death. The compound 4-bromobenzaldehyde N-(2,6-dimethylphenylsemicarbazone (EGA has been previously shown to protect cells from various bacterial protein toxins which deliver their enzymatic subunits from acidic endosomes to the cytosol, including Bacillus anthracis lethal toxin and the binary clostridial actin ADP-ribosylating toxins C2, iota and Clostridium difficile binary toxin (CDT. Here, we demonstrate that EGA also protects human cells from diphtheria toxin by inhibiting the pH-dependent translocation of DTA across cell membranes. The results suggest that EGA might serve for treatment and/or prevention of the severe disease diphtheria.

  14. Ageing of atrazine in manure amended soils assessed by bioavailability to Pseudomonas sp. strain ADP

    DEFF Research Database (Denmark)

    Glæsner, Nadia; Bælum, Jacob; Strobel, Bjarne W.;

    2014-01-01

    bacteria Pseudomonas sp. strain ADP. Throughout an ageing period of 90 days bioavailability was investigated at days 1, 10, 32, 60 and 90, where ~108 cells g−1 of the ADP strain was inoculated to the 14C-atrazine exposed soil and 14CO2 was collected over 7 days as a measure of mineralized atrazine. Even......Animal manure is applied to agricultural land in areas of high livestock production. In the present study, we evaluated ageing of atrazine in two topsoils with and without addition of manure and in one subsoil. Ageing was assessed as the bioavailability of atrazine to the atrazine mineralizing...... though the bioavailable residue decreased in all of the three soils as time proceeded, we found that ageing occurred faster in the topsoils rich in organic carbon than in subsoil. For one topsoil rich in organic carbon content, Simmelkær, we observed a higher degree of ageing when treated with manure...

  15. Ribosylated BSA Monomer is Severely Toxic to SH-SY5Y Cells%核糖糖基化BSA单体对SH-SY5Y细胞的毒性明显

    Institute of Scientific and Technical Information of China (English)

    魏艳; 王玉婧; 吴蓓蓓; 张英豪; 赫荣乔

    2016-01-01

    Oligomers,rather than polymers and fibrils,of protein aggregates are thought to be cytotoxic,which is a milestone in the study of protein misfolding and aggregation.Abnormally high level of uric ribose in type 2 diabetic patients and ribosylated animal models indicate that diabetes is not only correlated with metabolic dysfunction in glucose but also ribose.Here,using ribosylation of bovine serum albumin (BSA),we show that ribosylated BSA aggregates and proceeds from a monomer and onto an oligomer and polymer,observed with fluorescence spectrophotometer,atomic force microscopy,transmission electron microscopy and size exclusion chromatography.Ribosylated monomer showed severely cytotoxic to SH-SY5Y cells (a human neuroblastoma cell line) under the observations by assays of CCK-8,LDH activity,TUNEL staining,caspase-3 activity and flow-cytometry,whereas ribosylated oligomer and polymer did not.The cytotoxic effect of the ribosylated monomer likely occurs by inducing neuronal apoptosis through activation of the receptor of AGEs (RAGE) associated with mitogen-activated protein kinases (MAPK) pathways.%研究显示,蛋白质异常修饰形成的寡聚体,与其多聚体、淀粉样纤维相比,具有更强的细胞毒性.这一发现被认为是蛋白质错误折叠和聚集研究领域中的重要进展.蛋白质的异常修饰如还原糖的非酶糖基化,是糖尿病最基本的病理特征.2型糖尿病患者尿液中的核糖浓度显著升高,表明糖尿病不仅与葡萄糖代谢紊乱相关,同时也与核糖代谢失调相关.以牛血清白蛋白(BSA)为研究对象,通过荧光分光光度计检测、原子力显微镜、透射电子显微镜观察以及分子排阻色谱分离,观察到核糖糖基化能够诱导BSA聚集,从单体、寡聚体逐渐形成多聚体.通过CCK-8 Kit、乳酸脱氢酶细胞活性检测、TUNEL染色、caspase-3活性检测以及流式细胞检测等方法,发现核糖糖基化的BSA单体对SH-SY5Y细胞(人神经母细胞瘤

  16. Bacterial Enzymes and Antibiotic Resistance- Oral Presentation

    Energy Technology Data Exchange (ETDEWEB)

    Maltz, Lauren [SLAC National Accelerator Lab., Menlo Park, CA (United States)

    2015-08-25

    By using protein crystallography and X-ray diffraction, structures of bacterial enzymes were solved to gain a better understanding of how enzymatic modification acts as an antibacterial resistance mechanism. Aminoglycoside phosphotransferases (APHs) are one of three aminoglycoside modifying enzymes that confer resistance to the aminoglycoside antibiotics via enzymatic modification, rendering many drugs obsolete. Specifically, the APH(2”) family vary in their substrate specificities and also in their preference for the phosphate donor (ADP versus GDP). By solving the structures of members of the APH(2”) family of enzymes, we can see how domain movements are important to their substrate specificity. Our structure of the ternary complex of APH(2”)-IIIa with GDP and kanamycin, when compared to the known structures of APH(2”)-IVa, reveals that there are real physical differences between these two enzymes, a structural finding that explains why the two enzymes differ in their preferences for certain aminoglycosides. Another important group of bacterial resistance enzymes are the Class D β-lactamases. Oxacillinase carbapenemases (OXAs) are part of this enzyme class and have begun to confer resistance to ‘last resort’ drugs, most notably carbapenems. Our structure of OXA-143 shows that the conformational flexibility of a conserved hydrophobic residue in the active site (Val130) serves to control the entry of a transient water molecule responsible for a key step in the enzyme’s mechanism. Our results provide insight into the structural mechanisms of these two different enzymes.

  17. Cloning and expression of cDNA for human poly(ADP-ribose)polymerase

    International Nuclear Information System (INIS)

    cDNAs encoding poly(ADP-ribose) polymerase from a human hepatoma λgt11 cDNA library were isolated by immunological screening. One insert of 1.3 kilobases (kb) consistently hybridized on RNA gel blots to an mRNA species of 3.6-3.7 kb, which is consistent with the size of RNA necessary to code for the polymerase protein (116 kDa). This insert was subsequently used in both in vitro hybrid selection and hybrid-arrested translation studies. An mRNA species from HeLa cells of 3.6-3.7 kb was selected that was translated into a 116-kDa protein, which was selectively immunoprecipitated with anti-poly(ADP-ribose) polymerase. To confirm that the 1.3-kb insert from λgt11 encodes for poly(ADP-ribose) polymerase, the insert was used to screen a 3- to 4-kb subset of a transformed human fibroblast cDNA library in the Okayama-Berg vector. One of these vectors was tested in transient transfection experiments in COS cells. This cDNA insert contained the complete coding sequence for polymerase. Using pcD-p(ADPR)P as probe, it was observed that the level of poly(ADP-ribose) polymerase mRNA was elevated at 5 and 7 hr of S phase of the HeLa cell cycle, but was unaltered when artificial DNA strand breaks are introduced in HeLa cells by alkylating agents

  18. The Effect of Agricultural Development Project (ADP on the Rural Farmers in Adamawa State, Nigeria

    Directory of Open Access Journals (Sweden)

    Umar Adamu Madu

    2012-09-01

    Full Text Available Majority of communities in Nigeria are rural dwellers and agrarian by occupation. Development strategy for a country whose rural population are mainly farmers cannot be achieved without first sustained growth in rural income and standard of living primarily from agriculture. It was based on this that the state wide Agricultural Development Project (ADP was established to raise productivity, income and standard of living of rural farmers in Nigeria. This study assesses the effect of the ADP activities on the wellbeing of the rural farmers in Adamawa State, Nigeria. Data for this study were collect on annual crop output, annual income, farm size, use of improved technology, access to credit among farmers, farmers’ training and rural infrastructure development. The data were sourced using structured questionnaire and personal interviews. The statistical analysis used to determine the effect to the project on the participating farmers include, descriptive statistics and comparability test for difference (T-test analysis. The results indicates that Adamawa ADP had positive and significant impact on rural farmers productivity, income, access to credit, standard of living as measured by assets ownership. However, the project did not have significant impact on the rural infrastructure, adoption of improved technologies and farm sizes, even though the change from before and after ADP activities was positive. The study recommends that much attention should be paid to the provision of rural infrastructure and the needed improved technologies. The study also recommends that the two tiers of government in Nigeria should adequately fund the project to efficiently cope with its responsibility of developing the rural sector.

  19. D.C. electrical conductivity measurements on ADP single crystals added with simple organic compounds

    Indian Academy of Sciences (India)

    A Anne Assencia; C Mahadevan

    2005-08-01

    Pure and impurity added (with urea and thiourea) ADP single crystals were grown by the free evaporation method. D.C. electrical conductivity measurements were carried out along both the unique axis and perpendicular directions at various temperatures ranging from 40–150°C by the conventional two-probe method. Activation energies were also determined. The present study indicates that the conductivity increases with the increase in impurity concentration and temperature.

  20. Cloning and expression of cDNA for human poly(ADP-ribose)polymerase

    Energy Technology Data Exchange (ETDEWEB)

    Alkhatib, H.M.; Chen, D.; Cherney, B.; Bhatia, K.; Notario, V.; Giri, C.; Stein, G.; Slattery, E.; Roeder, R.G.; Smulson, M.E.

    1987-03-01

    cDNAs encoding poly(ADP-ribose) polymerase from a human hepatoma lambdagt11 cDNA library were isolated by immunological screening. One insert of 1.3 kilobases (kb) consistently hybridized on RNA gel blots to an mRNA species of 3.6-3.7 kb, which is consistent with the size of RNA necessary to code for the polymerase protein (116 kDa). This insert was subsequently used in both in vitro hybrid selection and hybrid-arrested translation studies. An mRNA species from HeLa cells of 3.6-3.7 kb was selected that was translated into a 116-kDa protein, which was selectively immunoprecipitated with anti-poly(ADP-ribose) polymerase. To confirm that the 1.3-kb insert from lambdagt11 encodes for poly(ADP-ribose) polymerase, the insert was used to screen a 3- to 4-kb subset of a transformed human fibroblast cDNA library in the Okayama-Berg vector. One of these vectors was tested in transient transfection experiments in COS cells. This cDNA insert contained the complete coding sequence for polymerase. Using pcD-p(ADPR)P as probe, it was observed that the level of poly(ADP-ribose) polymerase mRNA was elevated at 5 and 7 hr of S phase of the HeLa cell cycle, but was unaltered when artificial DNA strand breaks are introduced in HeLa cells by alkylating agents.

  1. Better detection of platelet aggregation in patients with metabolic syndrome using epinephrine and ADP

    OpenAIRE

    Perez-Campos-Mayoral, Laura; Pérez-Campos,Eduardo; Zenteno, Edgar; Majluf-Cruz, Abraham; Perez-Ortega, Eduardo; Matias-Pérez, Diana; Rodal-Canales, Francisco J; Martínez-Cruz, Ruth; Pina-Canseco, Socorro; Reyes Franco, Miguel Angel; Mayoral Andrade, Gabriel; Hernández, Pedro; Gallegos, Belem

    2014-01-01

    Background Patients with metabolic syndrome (MS) often have increased platelet aggregation. In order to determine which concentration detects a higher level of platelet aggregation in patients with MS, the agonists ADP and epinephrine were compared. Methods The study included 56 subjects with MS and 53 healthy subjects. Blood pressure, weight, body-mass index, and hip-to-waist ratio were collected from all subjects. Insulin, glucose, total serum cholesterol, HDL-C, LDL-C, total triglycerides,...

  2. PolyADP-ribose polymerase is a coactivator for AP-2-mediated transcriptional activation.

    OpenAIRE

    Kannan, P; Yu, Y; Wankhade, S; Tainsky, M A

    1999-01-01

    Overexpression of transcription factor AP-2 has been implicated in the tumorigenicity of the human teratocarcinoma cell lines PA-1 that contain an activated ras oncogene. Here we show evidence that overexpression of AP-2 sequesters transcriptional coactivators which results in self-inhibition. We identified AP-2-interacting proteins and determined whether these proteins were coactivators for AP-2-mediated transcription. One such interacting protein is polyADP-ribose polymerase (PARP). PARP su...

  3. Minocycline inhibits poly(ADP-ribose) polymerase-1 at nanomolar concentrations

    OpenAIRE

    Alano, Conrad C.; Kauppinen, Tiina M; Valls, Andreu Viader; Swanson, Raymond A.

    2006-01-01

    Poly(ADP-ribose) polymerase-1 (PARP-1), when activated by DNA damage, promotes both cell death and inflammation. Here we report that PARP-1 enzymatic activity is directly inhibited by minocycline and other tetracycline derivatives that have previously been shown to have neuroprotective and anti-inflammatory actions. These agents were evaluated by using cortical neuron cultures in which PARP-1 activation was induced by the genotoxic agents N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) or 3-morph...

  4. Treatment with insulin inhibits poly(ADP-ribose)polymerase activation in a rat model of endotoxemia

    OpenAIRE

    Horváth, Eszter M; Benk, Rita; Ger, Domonkos; Kiss, Levente; Szabó, Csaba

    2007-01-01

    In critically ill patients various conditions may lead to the activation of poly(ADP-ribose) polymerase (PARP). By promoting cellular energetic dysfunction, and by enhancing pro-inflammatory gene expression, PARP activation significantly contributes to the pathogenesis of shock. PARP activation is usually triggered by DNA strand breakage, which is typically the result of the overproduction of various reactive oxidant species. One of the pathophysiological conditions associated with PARP activ...

  5. Pyruvate dehydrogenase kinase isoform 2 activity limited and further inhibited by slowing down the rate of dissociation of ADP.

    Science.gov (United States)

    Bao, Haiying; Kasten, Shane A; Yan, Xiaohua; Roche, Thomas E

    2004-10-26

    Pyruvate dehydrogenase kinase 2 (PDK2) activity is enhanced by the dihydrolipoyl acetyltransferase core (E2 60mer) that binds PDK2 and a large number of its pyruvate dehydrogenase (E1) substrate. With E2-activated PDK2, K(+) at approximately 90 mM and Cl(-) at approximately 60 mM decreased the K(m) of PDK2 for ATP and competitive K(i) for ADP by approximately 3-fold and enhanced pyruvate inhibition. Comparing PDK2 catalysis +/- E2, E2 increased the K(m) of PDK2 for ATP by nearly 8-fold (from 5 to 39 microM), increased k(cat) by approximately 4-fold, and decreased the requirement for E1 by at least 400-fold. ATP binding, measured by a cold-trapping technique, occurred at two active sites with a K(d) of 5 microM, which equals the K(m) and K(d) of PDK2 for ATP measured in the absence of E2. During E2-aided catalysis, PDK2 had approximately 3 times more ADP than ATP bound at its active site, and the pyruvate analogue, dichloroacetate, led to 16-fold more ADP than ATP being bound (no added ADP). Pyruvate functioned as an uncompetitive inhibitor versus ATP, and inclusion of ADP transformed pyruvate inhibition to noncompetitive. At high pyruvate levels, pyruvate was a partial inhibitor but also induced substrate inhibition at high ATP levels. Our results indicate that, at physiological salt levels, ADP dissociation is a limiting step in E2-activated PDK2 catalysis, that PDK2.[ADP or ATP].pyruvate complexes form, and that PDK2.ATP.pyruvate.E1 reacts with PDK2.ADP.pyruvate accumulating. PMID:15491150

  6. ADP is a vasodilator component from Lasiodora sp. mygalomorph spider venom.

    Science.gov (United States)

    Horta, C C; Rezende, B A; Oliveira-Mendes, B B R; Carmo, A O; Capettini, L S A; Silva, J F; Gomes, M T; Chávez-Olórtegui, C; Bravo, C E S; Lemos, V S; Kalapothakis, E

    2013-09-01

    Members of the spider genus Lasiodora are widely distributed in Brazil, where they are commonly known as caranguejeiras. Lasiodora spider venom is slightly harmful to humans. The bite of this spider causes local pain, edema and erythema. However, Lasiodora sp. spider venom may be a source of important pharmacological tools. Our research group has described previously that Lasiodora sp. venom produces bradycardia in the isolated rat heart. In the present work, we sought to evaluate the vascular effect of Lasiodora sp. venom and to isolate the vasoactive compounds from the venom. The results showed that Lasiodora spider venom induced a concentration-dependent vasodilation in rat aortic rings, which was dependent on the presence of a functional endothelium and abolished by the nitric oxide synthase (NOS) inhibitor L-NAME. Western blot experiments revealed that the venom also increased endothelial NOS function by increasing phosphorylation of the Ser¹¹⁷⁷ residue. Assay-directed fractionation isolated a vasoactive fraction from Lasiodora sp. venom. Mass spectrometry (MS) and nuclear magnetic resonance (NMR) assays identified a mixture of two compounds: adenosine diphosphate (ADP, approximately 90%) and adenosine monophosphate (AMP, approximately 10%). The vasodilator effects of Lasiodora sp. whole venom, as well as ADP, were significantly inhibited by suramin, which is a purinergic P2-receptor antagonist. Therefore, the results of the present work indicate that ADP is a main vasodilator component of Lasiodora sp. spider venom.

  7. Yeast Mitochondrial Interactosome Model: Metabolon Membrane Proteins Complex Involved in the Channeling of ADP/ATP

    Directory of Open Access Journals (Sweden)

    Benjamin Clémençon

    2012-02-01

    Full Text Available The existence of a mitochondrial interactosome (MI has been currently well established in mammalian cells but the exact composition of this super-complex is not precisely known, and its organization seems to be different from that in yeast. One major difference is the absence of mitochondrial creatine kinase (MtCK in yeast, unlike that described in the organization model of MI, especially in cardiac, skeletal muscle and brain cells. The aim of this review is to provide a detailed description of different partner proteins involved in the synergistic ADP/ATP transport across the mitochondrial membranes in the yeast Saccharomyces cerevisiae and to propose a new mitochondrial interactosome model. The ADP/ATP (Aacp and inorganic phosphate (PiC carriers as well as the VDAC (or mitochondrial porin catalyze the import and export of ADP, ATP and Pi across the mitochondrial membranes. Aacp and PiC, which appear to be associated with the ATP synthase, consist of two nanomotors (F0, F1 under specific conditions and form ATP synthasome. Identification and characterization of such a complex were described for the first time by Pedersen and co-workers in 2003.

  8. Prevention of bacterial adhesion

    DEFF Research Database (Denmark)

    Klemm, Per; Vejborg, Rebecca Munk; Hancock, Viktoria

    2010-01-01

    Management of bacterial infections is becoming increasingly difficult due to the emergence and increasing prevalence of bacterial pathogens that are resistant to available antibiotics. Conventional antibiotics generally kill bacteria by interfering with vital cellular functions, an approach that ...

  9. Membrane coordination of receptors and channels mediating the inhibition of neuronal ion currents by ADP.

    Science.gov (United States)

    Gafar, Hend; Dominguez Rodriguez, Manuel; Chandaka, Giri K; Salzer, Isabella; Boehm, Stefan; Schicker, Klaus

    2016-09-01

    ADP and other nucleotides control ion currents in the nervous system via various P2Y receptors. In this respect, Cav2 and Kv7 channels have been investigated most frequently. The fine tuning of neuronal ion channel gating via G protein coupled receptors frequently relies on the formation of higher order protein complexes that are organized by scaffolding proteins and harbor receptors and channels together with interposed signaling components. However, ion channel complexes containing P2Y receptors have not been described. Therefore, the regulation of Cav2.2 and Kv7.2/7.3 channels via P2Y1 and P2Y12 receptors and the coordination of these ion channels and receptors in the plasma membranes of tsA 201 cells have been investigated here. ADP inhibited currents through Cav2.2 channels via both P2Y1 and P2Y12 receptors with phospholipase C and pertussis toxin-sensitive G proteins being involved, respectively. The nucleotide controlled the gating of Kv7 channels only via P2Y1 and phospholipase C. In fluorescence energy transfer assays using conventional as well as total internal reflection (TIRF) microscopy, both P2Y1 and P2Y12 receptors were found juxtaposed to Cav2.2 channels, but only P2Y1, and not P2Y12, was in close proximity to Kv7 channels. Using fluorescence recovery after photobleaching in TIRF microscopy, evidence for a physical interaction was obtained for the pair P2Y12/Cav2.2, but not for any other receptor/channel combination. These results reveal a membrane juxtaposition of P2Y receptors and ion channels in parallel with the control of neuronal ion currents by ADP. This juxtaposition may even result in apparent physical interactions between receptors and channels.

  10. THRIPS SPECIES (INSECTA: THYSANOPTERA OF ORNAMENTAL PLANTS FROM THE PARKS AND GREENHOUSES OF ADP PITESTI

    Directory of Open Access Journals (Sweden)

    Daniela Bărbuceanu

    2012-04-01

    Full Text Available The observations carried-out in 2008/2010 to ornamental plants from parks and greenhouses of ADP Pitesti relieve 12 species of thrips. One species of them, Frankliniella occidentalis was identified in greenhouses on Rosa sp., Dianthus sp. and Zantedeschia sp. In parks, the thrips species belong to 12 species, dominated by Frankliniella intonsa. All of them are polypfagous and divided in two throphic levels: primary and secondary consumers. The thrips species are mentioned for the first time in Romania on this host plant. In greenhouses are necessary intensive chemical treatments and methods of cultural hygiene to limit the F. occidentalis populations.

  11. Poly (ADP-ribose) polymerase inhibitor:an evolving paradigm in the treatment of prostate cancer

    Institute of Scientific and Technical Information of China (English)

    Jingsong Zhang

    2014-01-01

    Recent phase I studies have reported single-agent activities of poly (ADP-ribose) polymerase (PARP) inhibitor in sporadic and in BRCA-mutant prostate cancers. Two of the most common genetic alterations in prostate cancer, ETS gene rearrangement and loss of PTEN, have been linked to increased sensitivity to PARP inhibitor in preclinical models. Emerging evidence also suggests that PARP1 plays an important role in mediating the transcriptional activities of androgen receptor (AR) and ETS gene rearrangement. In this article, the preclinical work and early-phase clinical trials in developing PARP inhibitor-based therapy as a new treatment paradigm for metastatic prostate cancer are reviewed.

  12. Poly (ADP-ribose polymerase inhibitor: an evolving paradigm in the treatment of prostate cancer

    Directory of Open Access Journals (Sweden)

    Jingsong Zhang

    2014-06-01

    Full Text Available Recent phase I studies have reported single-agent activities of poly (ADP-ribose polymerase (PARP inhibitor in sporadic and in BRCA-mutant prostate cancers. Two of the most common genetic alterations in prostate cancer, ETS gene rearrangement and loss of PTEN, have been linked to increased sensitivity to PARP inhibitor in preclinical models. Emerging evidence also suggests that PARP1 plays an important role in mediating the transcriptional activities of androgen receptor (AR and ETS gene rearrangement. In this article, the preclinical work and early-phase clinical trials in developing PARP inhibitor-based therapy as a new treatment paradigm for metastatic prostate cancer are reviewed.

  13. Import of ADP/ATP carrier into mitochondria: two receptors act in parallel

    OpenAIRE

    1990-01-01

    We have identified the yeast homologue of Neurospora crassa MOM72, the mitochondrial import receptor for the ADP/ATP carrier (AAC), by functional studies and by cDNA sequencing. Mitochondria of a yeast mutant in which the gene for MOM72 was disrupted were impaired in specific binding and import of AAC. Unexpectedly, we found a residual, yet significant import of AAC into mitochondria lacking MOM72 that occurred via the receptor MOM19. We conclude that both MOM72 and MOM19 can direct AAC into ...

  14. Cloning and Expression Pattern of a Gene Encoding a Putative Plastidic ATP/ADP Transporter from Helianthus tuberosus L.

    Institute of Scientific and Technical Information of China (English)

    Kun MENG; Tuan-Jie CHANG; Xiang LIU; Song-Biao CHEN; Yong-Qin WANG; Ai-Jun SUN; Hong-Lin XU; Xiao-Li WEI; Zhen ZHU

    2005-01-01

    Herein, we report the cloning and molecular characterization of a full cDNA encoding a putative plastidic ATP/ADP transporter, designated HtAATP, for Helianthus tuberosus L. The ATP/ADP translocator protein was isolated from the tuber-cDNA library of H. tuberosus for the first time. The predicted HtAATP protein was judged as a plastidic ATP/ADP translocator protein from its high homology at the amino acid sequence level to the two Arabidopsis thaliana plastidic ATP/ADP translocator proteins AATP1 and AATP2 (84.8% and 79.9% identity, respectively). Amino acid sequence analysis of the primary structure of HtAATP revealed that it belonged to the plastidic ATP/ADP transporter family. Hydropathy prediction indicated that HtAATP gene product is a highly hydrophobic membrane protein that contains 10 transmembrane domains to form a spanning topology. Southern blotting analysis showed that the HtAATP gene is a single-copy gene in the H. tuberosus genome. Tissue distribution analysis showed that the HtAATP gene is prominently expressed in sink tissues. A stable expression pattern in tubers at different developmental stages implies an active involvement of HtAATP during carbohydrate formation.

  15. Prevention of bacterial adhesion

    DEFF Research Database (Denmark)

    Klemm, Per; Vejborg, Rebecca Munk; Hancock, Viktoria

    2010-01-01

    that imposes selection pressure for resistant bacteria. New approaches are urgently needed. Targeting bacterial virulence functions directly is an attractive alternative. An obvious target is bacterial adhesion. Bacterial adhesion to surfaces is the first step in colonization, invasion, and biofilm formation....... As such, adhesion represents the Achilles heel of crucial pathogenic functions. It follows that interference with adhesion can reduce bacterial virulence. Here, we illustrate this important topic with examples of techniques being developed that can inhibit bacterial adhesion. Some of these will become...

  16. Coexistence of ferroelectric and antiferroelectric microregions in the paraelectric phase of NH4H2PO4 (ADP)

    International Nuclear Information System (INIS)

    Ammonium dihydrogen phosphate NH4H2PO4 (ADP) is an antiferroelectric (AFE) compound belonging to the KDP-type family of hydrogen-bonded ferroelectrics. Recent ab initio results have shown that the optimization of the N-H-O bridges in ADP leads to the stabilization of the AFE state over a FE one. However, electron spin probe measurements have suggested that microregions of both phases may coexist above the critical antiferroelectric-paraelectric transition temperature. We have performed first principles studies of the energetics and relative stability of different AFE and FE defects embedded in a paraelectric (PE) matrix of ADP. Our analysis indicates that FE and AFE clusters are stable and may coexist in the PE phase, thus confirming the above suggestion.

  17. Nucleolin down-regulation is involved in ADP-induced cell cycle arrest in S phase and cell apoptosis in vascular endothelial cells.

    Directory of Open Access Journals (Sweden)

    Wenmeng Wang

    Full Text Available High concentration of extracellular ADP has been reported to induce cell apoptosis, but the molecular mechanisms remain not fully elucidated. In this study, we found by serendipity that ADP treatment of human umbilical vein endothelial cells (HUVEC and human aortic endothelial cells (HAEC down-regulated the protein level of nucleolin in a dose- and time-dependent manner. ADP treatment did not decrease the transcript level of nucloelin, suggesting that ADP might induce nucleolin protein degradation. HUVEC and HAEC expressed ADP receptor P2Y13 receptor, but did not express P2Y1 or P2Y12 receptors. However, P2Y1, 12, 13 receptor antagonists MRS2179, PSB0739, MRS2211 did not inhibit ADP-induced down-regulation of nucleolin. Moreover, MRS2211 itself down-regulated nucleolin protein level. In addition, 2-MeSADP, an agonist for P2Y1, 12 and 13 receptors, did not down-regulate nucleolin protein. These results suggested that ADP-induced nucleolin down-regulation was not due to the activation of P2Y1, 12, or 13 receptors. We also found that ADP treatment induced cell cycle arrest in S phase, cell apoptosis and cell proliferation inhibition via nucleolin down-regulation. The over-expression of nucleolin by gene transfer partly reversed ADP-induced cell cycle arrest, cell apoptosis and cell proliferation inhibition. Furthermore, ADP sensitized HUVEC to cisplatin-induced cell death by the down-regulation of Bcl-2 expression. Taken together, we found, for the first time to our knowledge, a novel mechanism by which ADP regulates cell proliferation by induction of cell cycle arrest and cell apoptosis via targeting nucelolin.

  18. Unraveling the activation mechanism of the potato tuber ADP-glucose pyrophosphorylase.

    Directory of Open Access Journals (Sweden)

    Carlos M Figueroa

    Full Text Available ADP-glucose pyrophosphorylase regulates the synthesis of glycogen in bacteria and of starch in plants. The enzyme from plants is mainly activated by 3-phosphoglycerate and is a heterotetramer comprising two small and two large subunits. Here, we found that two highly conserved residues are critical for triggering the activation of the potato tuber ADP-glucose pyrophosphorylase, as shown by site-directed mutagenesis. Mutations in the small subunit, which bears the catalytic function in this potato tuber form, had a more dramatic effect on disrupting the allosteric activation than those introduced in the large subunit, which is mainly modulatory. Our results strongly agree with a model where the modified residues are located in loops responsible for triggering the allosteric activation signal for this enzyme, and the sensitivity to this activation correlates with the dynamics of these loops. In addition, previous biochemical data indicates that the triggering mechanism is widespread in the enzyme family, even though the activator and the quaternary structure are not conserved.

  19.  Poly(ADP-ribose polymerase (PARP inhibitors in BRCA1/2 cancer therapy

    Directory of Open Access Journals (Sweden)

    Katarzyna Kluzek

    2012-06-01

    Full Text Available  A majority of currently used anticancer drugs belong to a group of chemical agents that damage DNA. The efficiency of the treatment is limited by effective DNA repair systems functioning in cancer cells. Many chemotherapeutic compounds cause strong systemic toxicity. Therefore, there is still a need for new anticancer agents which are less toxic for nontransformed cells and selectively kill cancer cells. One of the most promising molecular targets in cancer therapy is poly(ADP-ribose polymerases (PARP. PARP play an essential role in repairing DNA strand breaks. Small molecule inhibitors of these enzymes have been developed and have proved to be extremely toxic for cancer cells that lack the functional BRCA1 and BRCA2 proteins that are involved in homologous recombination, a complex repair mechanism of DNA double strand breaks. Mutations in BRCA1/2 genes are associated with genetically inherited breast and ovarian cancers. Therefore PARP inhibitors may prove to be very effective and selective in the treatment of these cancer types. This review is focused on the function of BRCA1/2 proteins and poly(ADP-ribose polymerases in DNA repair systems, especially in the homologous recombination process. A short history of the studies that led to synthesis of high specificity small molecule PARP inhibitors is also presented, as well as the results of clinical trials concerning the most effective PARP inhibitors in view of their potential application in oncological treatment, particularly breast cancers.

  20. A conserved loop in polynucleotide phosphorylase (PNPase) essential for both RNA and ADP/phosphate binding.

    Science.gov (United States)

    Carzaniga, Thomas; Mazzantini, Elisa; Nardini, Marco; Regonesi, Maria Elena; Greco, Claudio; Briani, Federica; De Gioia, Luca; Dehò, Gianni; Tortora, Paolo

    2014-02-01

    Polynucleotide phosphorylase (PNPase) reversibly catalyzes RNA phosphorolysis and polymerization of nucleoside diphosphates. Its homotrimeric structure forms a central channel where RNA is accommodated. Each protomer core is formed by two paralogous RNase PH domains: PNPase1, whose function is largely unknown, hosts a conserved FFRR loop interacting with RNA, whereas PNPase2 bears the putative catalytic site, ∼20 Å away from the FFRR loop. To date, little is known regarding PNPase catalytic mechanism. We analyzed the kinetic properties of two Escherichia coli PNPase mutants in the FFRR loop (R79A and R80A), which exhibited a dramatic increase in Km for ADP/Pi binding, but not for poly(A), suggesting that the two residues may be essential for binding ADP and Pi. However, both mutants were severely impaired in shifting RNA electrophoretic mobility, implying that the two arginines contribute also to RNA binding. Additional interactions between RNA and other PNPase domains (such as KH and S1) may preserve the enzymatic activity in R79A and R80A mutants. Inspection of enzyme structure showed that PNPase has evolved a long-range acting hydrogen bonding network that connects the FFRR loop with the catalytic site via the F380 residue. This hypothesis was supported by mutation analysis. Phylogenetic analysis of PNPase domains and RNase PH suggests that such network is a unique feature of PNPase1 domain, which coevolved with the paralogous PNPase2 domain.

  1. Analysis of current situation and perspective of digital preservation in the Social Science Data Archives (ADP

    Directory of Open Access Journals (Sweden)

    Irena Vipavc Brvar

    2011-01-01

    Full Text Available Social science data archives have begun to evolve in 1960’s, thus being one of the pioneers of digital preservation. The Slovenian Data Archive (ADP follows and adjusts best practices that have emerged in this specific field. Its tasks comprise acquisition,processing and long term preservation of valuable primary research data, as well as providing accessibility to the content for users. As in other areas of digital preservation much attention is paid to meeting the requirements of long term preservation as specified in the OAIS (Opean Archival Information System standard, and the use of technological solutions, proposed metadata standards, strategies and policies, and best practices. The purpose of this paper is to present procedures in ADP, to compare them with selected solutions of organizations with the similar mission, and to suggest improvements.Examples are taken from partner organizations within the CESSDA data archives association. These organizations (especially those from the USA, Great Britain and the Netherlands have developed solutions in the field of digital preservation to the highest degree. The paper also reflects on agreed guidelines of reliable services for the archives of research data, which arose in the frame of the FP7 project with the same title (i.e. CESSDA PPP. The results of analysis are useful for all are seeking practical solutions to similar problems.

  2. Ageing of atrazine in manure amended soils assessed by bioavailability to Pseudomonas sp. strain ADP.

    Science.gov (United States)

    Glæsner, Nadia; Bælum, Jacob; Strobel, Bjarne W; Jacobsen, Carsten S

    2014-04-01

    Animal manure is applied to agricultural land in areas of high livestock production. In the present study, we evaluated ageing of atrazine in two topsoils with and without addition of manure and in one subsoil. Ageing was assessed as the bioavailability of atrazine to the atrazine mineralizing bacteria Pseudomonas sp. strain ADP. Throughout an ageing period of 90 days bioavailability was investigated at days 1, 10, 32, 60 and 90, where ~10(8) cells g(-1) of the ADP strain was inoculated to the (14)C-atrazine exposed soil and (14)CO2 was collected over 7 days as a measure of mineralized atrazine. Even though the bioavailable residue decreased in all of the three soils as time proceeded, we found that ageing occurred faster in the topsoils rich in organic carbon than in subsoil. For one topsoil rich in organic carbon content, Simmelkær, we observed a higher degree of ageing when treated with manure. Contrarily, sorption experiments showed less sorption to Simmelkær treated with manure than the untreated soil indicating that sorption processes are not the only mechanisms of ageing. The other topsoil low in organic carbon content, Ringe, showed no significant difference in ageing between the manure-treated and untreated soil. The present study illustrates that not simply the organic carbon content influences adsorption and ageing of atrazine in soil but the origin and composition of organic matter plays an important role.

  3. Inhalation of nitric oxide inhibits ADP-induced platelet aggregation and alpha-granule release.

    Science.gov (United States)

    Hagberg, I A; Sølvik, U Ø; Opdahl, H; Roald, H E; Lyberg, T

    1999-01-01

    To gather further information about the effects on blood platelet activation of in vivo exposure to nitric oxide (NO), platelet reactivity was studied in blood from healthy, non-smoking male volunteers before and after 30 min inhalation of 40 ppm NO. Whole blood was stimulated in vitro with adenosine diphosphate or thrombin receptor activation peptide (TRAP-6). In an ex vivo perfusion model, non-anticoagulated blood was exposed to immobilised collagen at arterial blood flow conditions (2600 s(-1)). Blood samples from both the in vitro and ex vivo experiments were stained with fluorochrome-labelled Annexin-V and antibodies against CD42a, CD45, CD49b, CD61, CD62P and fibrinogen, and analysed with a three-colour flow cytometry technique. NO inhalation reduced the platelet activation response to adenosine diphosphate (ADP) stimulation by decreasing platelet-platelet aggregation, alpha-granule release and platelet-leukocyte conjugate formation. TRAP-stimulated platelet activation, collagen-induced platelet activation and thrombus growth was unaffected by NO inhalation. We therefore suggest an ADP receptor inhibitor mode of action of inhaled NO, selective on the newly suggested G protein- and phospholipase C-coupled P2Y1 receptor. Our results demonstrate that blood platelet activation in healthy subjects is modulated by inhalation of NO in therapeutically relevant doses, although the clinical impact of our findings remains unclear. PMID:16801117

  4. Cloning and characterization of ADP-glucose pyrophosphorylase small subunit gene in Cyperus esculentus (yellow nutsedge).

    Science.gov (United States)

    Cheng, C; Hu, J; Zhi, Y; Su, J J; Zhang, X K; Huang, B Q

    2015-01-01

    ADP-glucose pyrophosphorylase (ADPGlcPPase) controls the first committed step of starch synthesis by catalyzing the biosynthesis of ADP-glucose from glucose-phosphate and ATP. It is a tetrameric protein consisting of two small and two large subunits. The small subunits have a catalytic function, while the large subunits regulate the enzyme activity. Cyperus esculentus (yellow nutsedge) is a perennial C4 plant grown from rhizomes and tubers. Previous studies on yellow nutsedge have mostly focused on the morphology and cultivation of tubers, their application in food, and biochemical analyses of the tubers. In this study, the gene encoding the ADPGlcPPase small subunit (CeAGPS) in yellow nutsedge was cloned and characterized. The full-length CeAGPS cDNA sequence contained an 81-bp 5'-untranslated region (UTR), a 188-bp 3'-UTR, and a 1539-bp open reading frame encoding 512-amino acid residues. The genomic sequence of CeAGPS comprises a nine exon-eight intron structure similar to the previously reported cotton and Arabidopsis thaliana AGPS genes. The deduced translation product of the CeAGPS gene contained a well-conserved catalytic domain and regulatory elements typical of plant AGPS. Reverse transcriptase polymerase chain reaction amplification of the target gene in various plant parts using gene-specific primers indicated that the expression of CeAGPS was most abundant in the tuber, and relatively lower in nutsedge roots. PMID:26782478

  5. Role of CD38, a cyclic ADP-ribosylcyclase, in morphine antinociception and tolerance.

    Science.gov (United States)

    Hull, Lynn C; Rabender, Christopher; Gabra, Bichoy H; Zhang, Fan; Li, Pin-Lan; Dewey, William L

    2010-09-01

    Our previous studies have demonstrated that an increase in intracellular levels of Ca(2+) in neurons is an important component of both the antinociception produced by morphine and morphine's tolerance. The present study tested the hypothesis that the Ca(2+) signaling second messenger, cyclic ADP-ribose (cADPR), derived from CD38 activation participates in morphine antinociception and tolerance. We first showed that morphine's antinociceptive potency was increased by the intracerebroventricular injection of CD38 substrate beta-NAD(+) in mice. Furthermore, morphine tolerance was reversed by intracerebroventricular administration of each of three different inhibitors of the CD38-cADPR-ryanodine receptor Ca(2+) signaling pathway. These inhibitors were the ADP-ribosylcyclase inhibitor nicotinamide, cADPR analog 8-bromo-cADPR, and a large dose of ryanodine (>50 muM) that blocks the ryanodine receptor. In CD38 gene knockout [CD38(-/-)] mice, the antinociceptive action of morphine was found to be less potent compared with wild-type (WT) mice, as measured by tail-flick response, hypothermia assay, and observations of straub tail. However, there was no difference in locomotor activation between CD38(-/-) and WT animals. It was also found that less tolerance to morphine developed in CD38(-/-) mice compared with WT animals. These results indicate that cADRP-ryanodine receptor Ca(2+) signaling associated with CD38 plays an important role in morphine tolerance. PMID:20551293

  6. P2Y12-ADP receptor antagonists: Days of future and past.

    Science.gov (United States)

    Laine, Marc; Paganelli, Franck; Bonello, Laurent

    2016-05-26

    Antiplatelet therapy is the cornerstone of the therapeutic arsenal in coronary artery disease. Thanks to a better understanding in physiology, pharmacology and pharmacogenomics huge progress were made in the field of platelet reactivity inhibition thus allowing the expansion of percutaneous coronary intervention. Stent implantation requires the combination of two antiplatelet agents acting in a synergistic way. Asprin inhibit the cyclo-oxygenase pathway of platelet activation while clopidogrel is a P2Y12 adenosine diphosphate (ADP)-receptor antagonist. This dual antiplatelet therapy has dramatically improved the prognosis of stented patients. However, due to pharmacological limitations of clopidogrel (interindividual variability in its biological efficacy, slow onset of action, mild platelet reactivity inhibition) ischemic recurrences remained high following stent implantation especially in acute coronary syndrome patients. Thus, more potent P2Y12-ADP receptor inhibitors were developped including prasugrel, ticagrelor and more recently cangrelor to overcome these pitfalls. These new agents reduced the rate of thrombotic events in acute coronary syndrome patients at the cost of an increased bleeding risk. The abundance in antiplatelet agents allow us to tailor our strategy based on the thrombotic/bleeding profile of each patient. Recently, the ACCOAST trial cast a doubt on the benefit of pre treatment in non-ST segment elevation acute coronary syndrome. The aim of the present review is to summarize the results of the main studies dealing with antiplatelet therapy in stented/acute coronary syndromes patients. PMID:27231519

  7. Phosphorylation of Thr18 and Ser20 of p53 in Ad-p53 – induced apoptosis

    OpenAIRE

    Nakamizo, Akira; Amano, Toshiyuko; Zhang, Wei; Zhang, Xin-qiao; Ramdas, Latha; Liu, Ta-Jen; Bekele, B. Nebiyou; Shono, Tadahisa; Sasaki, Tomio; Benedict, William F.; Sawaya, Raymond; Lang, Frederick F.

    2008-01-01

    The p53 protein plays a critical role in inducing cell cycle arrest or apoptosis. Because p53 is inactivated in human gliomas, restoring p53 function is a major focus of glioma therapy. The most clinically tested strategy for replacing p53 has been adenoviral-mediated p53 gene therapy (Ad-p53). In addition to their therapeutic implications, investigations into Ad-p53 provide model systems for understanding p53’s ability to induce cell cycle arrest versus apoptosis, particularly because wild-t...

  8. Crystallization and preliminary X-ray diffraction analysis of the amidase domain of allophanate hydrolase from Pseudomonas sp. strain ADP

    International Nuclear Information System (INIS)

    The amidase domain of the allophanate hydrolase AtzF from Pseudomonas sp. strain ADP has been crystallized and preliminary X-ray diffraction data have been collected. The allophanate hydrolase from Pseudomonas sp. strain ADP was expressed and purified, and a tryptic digest fragment was subsequently identified, expressed and purified. This 50 kDa construct retained amidase activity and was crystallized. The crystals diffracted to 2.5 Å resolution and adopted space group P21, with unit-cell parameters a = 82.4, b = 179.2, c = 112.6 Å, β = 106.6°

  9. Poly(ADP-ribose)--a unique natural polymer structural features, biological role and approaches to the chemical synthesis.

    Science.gov (United States)

    Drenichev, Mikhail S; Mikhailov, Sergey N

    2015-01-01

    Poly(ADP-ribose) (PAR) is a natural polymer, taking part in numerous important cellular processes. Several enzymes are involved in biosynthesis and degradation of PAR. One of them, poly(ADP-ribose)polymerase-1 (PARP-1) is considered to be a perspective target for the design of new drugs, affecting PAR metabolism. The structure of PAR was established by enzymatic hydrolysis and further analysis of the products, but total chemical synthesis of PAR hasn't been described yet. Several approaches have been developed on the way to chemical synthesis of this unique biopolymer.

  10. Postnatal Age Influences Hypoglycemia-induced Poly(ADP-ribose) Polymerase-1 Activation in the Brain Regions of Rats

    OpenAIRE

    Rao, Raghavendra; Sperr, Dustin; Ennis, Kathleen; Tran, Phu

    2009-01-01

    Poly(ADP-ribose) polymerase-1 (PARP-1) overactivation plays a significant role in hypoglycemia-induced brain injury in adult rats. To determine the influence of postnatal age on PARP-1 activation, developing and adult male rats were subjected to acute hypoglycemia of equivalent severity and duration. The expression of PARP-1 and its downstream effectors, apoptosis inducing factor (Aifm1), caspase 3 (Casp3), NF-κB (Nfkb1) and bcl-2 (Bcl2), and cellular poly(ADP-ribose) (PAR) polymer expression...

  11. Prediction of the Occurrence of the ADP-binding βαβ-fold in Proteins, Using an Amino Acid Sequence Fingerprint

    NARCIS (Netherlands)

    Wierenga, Rik K.; Terpstra, Peter; Hol, Wim G.J.

    1986-01-01

    An amino acid sequence "fingerprint” has been derived that can be used to test if a particular sequence will fold into a βαβ-unit with ADP-binding properties. It was deduced from a careful analysis of the known three-dimensional structures of ADP-binding βαβ-folds. This fingerprint is in fact a set

  12. GNOM-LIKE 2, Encoding an Adenosine Diphosphate-Ribosylation Factor-Guanine Nucleotide Exchange Factor Protein Homologous to GNOM and GNL1, is Essential for Pollen Germination in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Dong-Jie Jia; Xi Cao; Wei Wang; Xiao-Yun Tan; Xue-Qin Zhang; Li-Qun Chen; De Ye

    2009-01-01

    In flowering plants, male gametes are delivered to female gametophytes by pollen tubes. Although it is important for sexual plant reproduction, little is known about the genetic mechanism that controls pollen germination and pollen tube growth. Here we report the identification and characterization of two novel mutants, gnom-like 2.1 (gnl2-1) and gn12-2 in Arabidopsis thaliana, in which the pollen grains failed to germinate in vitro and in vivo. GNL2 encodes a protein homologous to the adenosine diphosphate-ribosylation factor-guanine nucleotide exchange factors, GNOM and GNL1 that are involved in endosomal recycling and endoplasmic reticulum-Golgi vesicular trafficking, it was prolifically expressed in pollen grains and pollen tubes. The results of the present study suggest that GNL2 plays an important role in pollen germination.

  13. A divergent ADP/ATP carrier in the hydrogenosomes of Trichomonas gallinae argues for an independent origin of these organelles.

    NARCIS (Netherlands)

    Tjaden, J.; Haferkamp, I.; Boxma, B.; Tielens, A.G.; Huynen, M.A.; Hackstein, J.H.P.

    2004-01-01

    The evolution of mitochondrial ADP and ATP exchanging proteins (AACs) highlights a key event in the evolution of the eukaryotic cell, as ATP exporting carriers were indispensable in establishing the role of mitochondria as ATP-generating cellular organelles. Hydrogenosomes, i.e. ATP- and hydrogen-ge

  14. 31P saturation transfer spectroscopy predicts differential intracellular macromolecular association of ATP and ADP in skeletal muscle.

    NARCIS (Netherlands)

    Nabuurs, C.I.H.C.; Huijbregts, B.; Wieringa, B.; Hilbers, C.W.; Heerschap, A.

    2010-01-01

    The kinetics of phosphoryl exchange involving ATP and ADP have been investigated successfully by in vivo (31)P magnetic resonance spectroscopy using magnetization transfer. However, magnetization transfer effects seen on the signals of ATP also could arise from intramolecular cross-relaxation. This

  15. 3'-O-(5-fluoro-2,4-dinitrophenyl)ADP ether and ATP ether. Affinity reagents for labeling ATPases.

    Science.gov (United States)

    Chuan, H; Wang, J H

    1988-09-15

    The affinity reagents 3'-O-(5-fluoro-2,4-dinitrophenyl)ADP ether (FDNP-ADP) and 3'-O-(5-fluoro-2,4-dinitrophenyl)ATP ether (FDNP-ATP) were synthesized and characterized. FDNP[14C]ADP was found to label the active site of mitochondrial F1-ATPase slowly at room temperature but with high specificity. F1 was effectively protected from the labeling reagent by ATP or ADP. An average number of 1.3 covalent label per F1 is sufficient for 100% inhibition of the ATPase. About 73% of the radioactive label was found covalently attached to beta subunits, 9% on alpha, practically none on gamma, delta, and epsilon. Cleavage of the labeled enzyme by pepsin and sequencing of the major radioactive peptide showed that the labeled amino acid residue in beta subunit was Lys beta 162. These results show that Lys beta 162 is indeed at the active site of F1 as assumed in the recently proposed models (Fry, D. C., Kuby, S. A., and Mildvan, A. S. (1986) Proc. Natl. Acad. Sci. U. S. A. 83, 907-911; Duncan, I. M., Parsonage, D., and Senior, A. E. (1986) FEBS Lett. 208, 1-6).

  16. Influence of the ADP/ATP ratio, 2-oxoglutarate and divalent ions on Azospirillum brasilense PII protein signalling.

    Science.gov (United States)

    Gerhardt, Edileusa C M; Araújo, Luíza M; Ribeiro, Ronny R; Chubatsu, Leda S; Scarduelli, Marcelo; Rodrigues, Thiago E; Monteiro, Rose A; Pedrosa, Fábio O; Souza, Emanuel M; Huergo, Luciano F

    2012-06-01

    Proteins belonging to the P(II) family coordinate cellular nitrogen metabolism by direct interaction with a variety of enzymes, transcriptional regulators and transporters. The sensing function of P(II) relies on its ability to bind the nitrogen/carbon signalling molecule 2-oxoglutarate (2-OG). In Proteobacteria, P(II) is further subject to reversible uridylylation according to the intracellular levels of glutamine, which reflect the cellular nitrogen status. A number of P(II) proteins have been shown to bind ADP and ATP in a competitive manner, suggesting that P(II) might act as an energy sensor. Here, we analyse the influence of the ADP/ATP ratio, 2-OG levels and divalent metal ions on in vitro uridylylation of the Azospirillum brasilense P(II) proteins GlnB and GlnZ, and on interaction with their targets AmtB, DraG and DraT. The results support the notion that the cellular concentration of 2-OG is a key factor governing occupation of the GlnB and GlnZ nucleotide binding sites by ATP or ADP, with high 2-OG levels favouring the occupation of P(II) by ATP. Both P(II) uridylylation and interaction with target proteins responded to the ADP/ATP ratio within the expected physiological range, supporting the concept that P(II) proteins might act as cellular energy sensors.

  17. ATP- and ADP-dnaA protein, a molecular switch in gene regulation.

    OpenAIRE

    Speck, C.; Weigel, C; Messer, W

    1999-01-01

    DnaA protein functions by binding to asymmetric 9mer DNA sites, the DnaA boxes. ATP-DnaA and ADP-DnaA bind to 9mer DnaA boxes with equal affinity, but only ATP-DnaA protein binds in addition to an as yet unknown 6mer site, the ATP-DnaA box AGATCT, or a close match to it. ATP-DnaA protein binding to ATP-DnaA boxes is restricted to sites located in close proximity to DnaA boxes, suggesting that protein-protein interaction is required for its stabilization. We show that ATP-DnaA represses dnaA t...

  18. Poly(ADP-Ribose) Polymerase in Cervical Cancer Pathogenesis: Mechanism and Potential Role for PARP Inhibitors.

    Science.gov (United States)

    Kotsopoulos, Ioannis C; Kucukmetin, Ali; Mukhopadhyay, Asima; Lunec, John; Curtin, Nicola J

    2016-05-01

    Treatment options for disease recurrence of women treated for locally advanced and advanced cervical cancer are very limited-largely palliative chemotherapy. The low efficacy of the currently available drugs raises the need for new targeted agents. Poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) inhibitors (PARPi) have emerged as a promising class of chemotherapeutic agents in cancers associated with defects in DNA repair. Their therapeutic potential in cervical cancer is currently being evaluated in 3 ongoing clinical trials. Here we review the available information regarding all the aspects of PARP in cervical intraepithelial neoplasia and invasive cervical cancer, from expression and the mechanism of action to the role of the polymorphisms in the pathogenesis of the disease, as well as the potential of the inhibitors. We finally propose a new unifying theory regarding the role of PARPs in the development of cervical carcinomas. PMID:26905326

  19. Regulatory properties of potato-Arabidopsis hybrid ADP-glucose pyrophosphorylase.

    Science.gov (United States)

    Ventriglia, Tiziana; Ballicora, Miguel A; Crevillén, Pedro; Preiss, Jack; Romero, José M

    2007-06-01

    In higher plants, ADP-glucose pyrophosphorylase (ADPGlc-PPase) is a heterotetrameric enzyme comprised of two small and two large subunits. Potato-Arabidopsis hybrid ADPGlc-PPases were generated and their regulatory properties analyzed. We show that ADPGlc-PPase subunits from two different species can interact, producing active enzymes with new regulatory properties. Depending on the subunit combinations, hybrid heterotetramers showed responses to allosteric effectors [3-phosphoglycerate (3-PGA) and Pi] in the micromolar or millimolar range. While hybrid potato small subunit (PSS) and the Arabidopsis large subunit APL1 showed an extremely sensitive response to 3-PGA and Pi, hybrid PSS/Arabidopsis APL2 was very insensitive to them. Intermediate responses were determined for other subunit combinations. PMID:17452341

  20. Structure-activity relationship of cyclic ADP-ribose, an update

    Institute of Scientific and Technical Information of China (English)

    Andreas H.Guse

    2013-01-01

    Cyclic ADP-ribose (cADPR) is a universal Ca2+ mobilizing second messenger in many different cell types and organisms.cADPR activates Ca2+ release from endo/sarcoplasmic reticulum via ryanodine receptors.In addition,Ca2+ entry secondary to Ca2+ depletion is at least one of the mechanisms in which cADPR triggers Ca2+ inflow,too.Analogues of cADPR have been prepared by chemical and chemo-enzymatic routes.Most of the analogues were analyzed for biological activity in intact or permeabilized Jurkat T cells (a human T-lymphoma cell line).As a systematic approach,analogues were grouped according to alterations in the base,the northern ribose,the southern ribose,the pyrophosphate backbone,or in complex modifications,comprising more than one part of the molecule.Biological activity of the analogues is reviewed,with special emphasis on Jurkat T cells.

  1. Poly (ADP-ribose) polymerase-1 gene polymorphism in various Chinese nationalities

    Institute of Scientific and Technical Information of China (English)

    Hairong Liang; Junli Shao; Yuting Gao; Linhua Liu; Juanxiu Dai; Yun He; Huanwen Tang

    2011-01-01

    Poly (ADP-ribose) polymerase-1 (PARP-1) can exacerbate ischemic brain injury and lessen ischemic neuronal death, which may be associated with PARP-1 polymorphisms. The present study investigated human PARP-1 gene polymorphisms in various Chinese nationalities, the results of which could potentially help in the treatment and prevention of neurologic diseases. Genetic polymorphisms of seven exons in the PARP-1 gene, in 898 Chinese Han, Buyi, Shui, Miao, and Zhuang subjects, were investigated by PCR-single-strand conformation polymorphism. A single-strand conformation polymorphism variant in exons 12, 13, 16, and 17 of the PARP-1 gene was identified in 148 people, with two stationary bands showing three degenerative single strands.Results showed that the PARP-1 gene polymorphisms exist in various nationalities, and may act as a biomarker for susceptibility to disease.

  2. The Role of Poly(ADP-ribose Polymerase-1 in Rheumatoid Arthritis

    Directory of Open Access Journals (Sweden)

    Samuel García

    2015-01-01

    Full Text Available Poly(ADP-ribose polymerase-1 (PARP-1 is a nuclear enzyme with a crucial role in the maintenance of genomic stability. In addition to the role of PARP-1 in DNA repair, multiple studies have also demonstrated its involvement in several inflammatory diseases, such as septic shock, asthma, atherosclerosis, and stroke, as well as in cancer. In these diseases, the pharmacological inhibition of PARP-1 has shown a beneficial effect, suggesting that PARP-1 regulates their inflammatory processes. In recent years, we have studied the role of PARP-1 in rheumatoid arthritis, as have other researchers, and the results have shown that PARP-1 has an important function in the development of this disease. This review summarizes current knowledge on the effects of PARP-1 in rheumatoid arthritis.

  3. Substrate binding properties of potato tuber ADP-glucose pyrophosphorylase as determined by isothermal titration calorimetry.

    Science.gov (United States)

    Cakir, Bilal; Tuncel, Aytug; Green, Abigail R; Koper, Kaan; Hwang, Seon-Kap; Okita, Thomas W; Kang, ChulHee

    2015-06-01

    Substrate binding properties of the large (LS) and small (SS) subunits of potato tuber ADP-glucose pyrophosphorylase were investigated by using isothermal titration calorimetry. Our results clearly show that the wild type heterotetramer (S(WT)L(WT)) possesses two distinct types of ATP binding sites, whereas the homotetrameric LS and SS variant forms only exhibited properties of one of the two binding sites. The wild type enzyme also exhibited significantly increased affinity to this substrate compared to the homotetrameric enzyme forms. No stable binding was evident for the second substrate, glucose-1-phosphate, in the presence or absence of ATPγS suggesting that interaction of glucose-1-phosphate is dependent on hydrolysis of ATP and supports the Theorell-Chance bi bi reaction mechanism.

  4. Substrate binding properties of potato tuber ADP-glucose pyrophosphorylase as determined by isothermal titration calorimetry.

    Science.gov (United States)

    Cakir, Bilal; Tuncel, Aytug; Green, Abigail R; Koper, Kaan; Hwang, Seon-Kap; Okita, Thomas W; Kang, ChulHee

    2015-06-01

    Substrate binding properties of the large (LS) and small (SS) subunits of potato tuber ADP-glucose pyrophosphorylase were investigated by using isothermal titration calorimetry. Our results clearly show that the wild type heterotetramer (S(WT)L(WT)) possesses two distinct types of ATP binding sites, whereas the homotetrameric LS and SS variant forms only exhibited properties of one of the two binding sites. The wild type enzyme also exhibited significantly increased affinity to this substrate compared to the homotetrameric enzyme forms. No stable binding was evident for the second substrate, glucose-1-phosphate, in the presence or absence of ATPγS suggesting that interaction of glucose-1-phosphate is dependent on hydrolysis of ATP and supports the Theorell-Chance bi bi reaction mechanism. PMID:25953126

  5. Pyruvate dehydrogenase kinase isoform 2 activity stimulated by speeding up the rate of dissociation of ADP.

    Science.gov (United States)

    Bao, Haiying; Kasten, Shane A; Yan, Xiaohua; Hiromasa, Yasuaki; Roche, Thomas E

    2004-10-26

    Pyruvate dehydrogenase kinase 2 (PDK2) activity is stimulated by NADH and NADH plus acetyl-CoA via the reduction and reductive acetylation of the lipoyl groups of the dihydrolipoyl acetyltransferase (E2) component. Elevated K(+) and Cl(-) were needed for significant stimulation. Stimulation substantially increased both k(cat) and the K(m) for ATP; the fractional stimulation increased with the level of ATP. With an E2 structure lacking the pyruvate dehydrogenase (E1) binding domain, stimulation of PDK2 was retained, the K(m) for E1 decreased, and the equilibrium dissociation constant for ATP increased but remained much lower than the K(m) for ATP. Stimulation of PDK2 activity greatly reduced the fraction of bound ADP. These results fit an ordered reaction mechanism with ATP binding before E1 and stimulation increasing the rate of dissociation of ADP. Conversion of all of the lipoyl groups in the E2 60mer to the oxidized form (E2(ox)) greatly reduced k(cat) and the K(m) of PDK2 for ATP. Retention over an extended period of time of a low portion of reduced lipoyl groups maintains E2 in a state that supported much higher PDK2 activity than short-term (5 min) reduction of a large portion of lipoyl groups of E2(ox), but reduction of E2(ox) produced a larger fold stimulation. Reduction and to a greater extent reductive acetylation increased PDK2 binding to E2; conversion to E2(ox) did not significantly hinder binding. We suggest that passing even limited reducing equivalents among lipoyl groups maintains E2 lipoyl domains in a conformation that aids kinase function. PMID:15491151

  6. ADP-flow velocity profile to interpret hydromorphological features of China's Yangtze Three-Gorges valley

    Institute of Scientific and Technical Information of China (English)

    CHEN Jing; CHEN Zhongyuan; XU Kaiqin; WEI Taoyuan; LI Maotian; WANG Zhanghua; Masataka Watanabe

    2005-01-01

    In late May and early June, 2002, a field investigation was conducted along the Three-Gorges valley of the upper Yangtze catchment by ADP (Acoustic Doppler Profile SONTEK-500). Data obtained when surveying were accompanied with discharge of 1000 m) and shallower water depth (50 m) and U-shaped river-channel morphology. Mapping the river cross-section area at those sites can determine that smaller cross-section area accelerates the flow velocity. From Wanxian to Fengjie, the average flow velocity ranging from 3.0 to 4.5 m/s is in-phase with the water depth. The high-flow velocity is associated with narrower river-channel, where V-shaped gorges valley occurs with small cross-section area. Further downstream from Fengjie to Zigui, the low flow velocity is linked to deep river channel characterized by W-shaped valley morphology of large cross-section area, in general. The average flow velocity is 2.5―3.5 m/s, and maximum can reach 6.0 m/s near Wu-Gorge. Our survey had also detected a slow-flow velocity (mostly 100 m; maximum) in the gorges valley (30―40 m below the present mean sea level). This contrasts to the relative shallow water river-channel above Fengjie, i.e. 20―30 m in general and 50―60 m, maximum at gorges site. The present ADP investigation displays the hydromorphological feature in the Three-Gorges valley, and most importantly, it accumulates invaluable dataset for the post-dam study in the near future.

  7. Vimentin in Bacterial Infections

    Directory of Open Access Journals (Sweden)

    Tim N. Mak

    2016-04-01

    Full Text Available Despite well-studied bacterial strategies to target actin to subvert the host cell cytoskeleton, thus promoting bacterial survival, replication, and dissemination, relatively little is known about the bacterial interaction with other components of the host cell cytoskeleton, including intermediate filaments (IFs. IFs have not only roles in maintaining the structural integrity of the cell, but they are also involved in many cellular processes including cell adhesion, immune signaling, and autophagy, processes that are important in the context of bacterial infections. Here, we summarize the knowledge about the role of IFs in bacterial infections, focusing on the type III IF protein vimentin. Recent studies have revealed the involvement of vimentin in host cell defenses, acting as ligand for several pattern recognition receptors of the innate immune system. Two main aspects of bacteria-vimentin interactions are presented in this review: the role of vimentin in pathogen-binding on the cell surface and subsequent bacterial invasion and the interaction of cytosolic vimentin and intracellular pathogens with regards to innate immune signaling. Mechanistic insight is presented involving distinct bacterial virulence factors that target vimentin to subvert its function in order to change the host cell fate in the course of a bacterial infection.

  8. Nucleotide-Induced Conformational Changes in Escherichia coli DnaA Protein Are Required for Bacterial ORC to Pre-RC Conversion at the Chromosomal Origin.

    Science.gov (United States)

    Saxena, Rahul; Vasudevan, Sona; Patil, Digvijay; Ashoura, Norah; Grimwade, Julia E; Crooke, Elliott

    2015-01-01

    DnaA oligomerizes when bound to origins of chromosomal replication. Structural analysis of a truncated form of DnaA from Aquifex aeolicus has provided insight into crucial conformational differences within the AAA+ domain that are specific to the ATP- versus ADP- bound form of DnaA. In this study molecular docking of ATP and ADP onto Escherichia coli DnaA, modeled on the crystal structure of Aquifex aeolicus DnaA, reveals changes in the orientation of amino acid residues within or near the vicinity of the nucleotide-binding pocket. Upon limited proteolysis with trypsin or chymotrypsin ADP-DnaA, but not ATP-DnaA generated relatively stable proteolytic fragments of various sizes. Examined sites of limited protease susceptibility that differ between ATP-DnaA and ADP-DnaA largely reside in the amino terminal half of DnaA. The concentration of adenine nucleotide needed to induce conformational changes, as detected by these protease susceptibilities of DnaA, coincides with the conversion of an inactive bacterial origin recognition complex (bORC) to a replication efficient pre-replication complex (pre-RC) at the E. coli chromosomal origin of replication (oriC). PMID:26610483

  9. Nucleotide-Induced Conformational Changes in Escherichia coli DnaA Protein Are Required for Bacterial ORC to Pre-RC Conversion at the Chromosomal Origin

    Directory of Open Access Journals (Sweden)

    Rahul Saxena

    2015-11-01

    Full Text Available DnaA oligomerizes when bound to origins of chromosomal replication. Structural analysis of a truncated form of DnaA from Aquifex aeolicus has provided insight into crucial conformational differences within the AAA+ domain that are specific to the ATP- versus ADP- bound form of DnaA. In this study molecular docking of ATP and ADP onto Escherichia coli DnaA, modeled on the crystal structure of Aquifex aeolicus DnaA, reveals changes in the orientation of amino acid residues within or near the vicinity of the nucleotide-binding pocket. Upon limited proteolysis with trypsin or chymotrypsin ADP-DnaA, but not ATP-DnaA generated relatively stable proteolytic fragments of various sizes. Examined sites of limited protease susceptibility that differ between ATP-DnaA and ADP-DnaA largely reside in the amino terminal half of DnaA. The concentration of adenine nucleotide needed to induce conformational changes, as detected by these protease susceptibilities of DnaA, coincides with the conversion of an inactive bacterial origin recognition complex (bORC to a replication efficient pre-replication complex (pre-RC at the E. coli chromosomal origin of replication (oriC.

  10. Interfering with bacterial gossip

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

    2011-01-01

    defense. Antibiotics exhibit a rather limited effect on biofilms. Furthermore, antibiotics have an ‘inherent obsolescence’ because they select for development of resistance. Bacterial infections with origin in bacterial biofilms have become a serious threat in developed countries. Pseudomonas aeruginosa...... that appropriately target bacteria in their relevant habitat with the aim of mitigating their destructive impact on patients. In this review we describe molecular mechanisms involved in “bacterial gossip” (more scientifically referred to as quorum sensing (QS) and c-di-GMP signaling), virulence, biofilm formation...

  11. Bacterial Wound Culture

    Science.gov (United States)

    ... Home Visit Global Sites Search Help? Bacterial Wound Culture Share this page: Was this page helpful? Also known as: Aerobic Wound Culture; Anaerobic Wound Culture Formal name: Culture, wound Related ...

  12. Bacterial surface adaptation

    Science.gov (United States)

    Utada, Andrew

    2014-03-01

    Biofilms are structured multi-cellular communities that are fundamental to the biology and ecology of bacteria. Parasitic bacterial biofilms can cause lethal infections and biofouling, but commensal bacterial biofilms, such as those found in the gut, can break down otherwise indigestible plant polysaccharides and allow us to enjoy vegetables. The first step in biofilm formation, adaptation to life on a surface, requires a working knowledge of low Reynolds number fluid physics, and the coordination of biochemical signaling, polysaccharide production, and molecular motility motors. These crucial early stages of biofilm formation are at present poorly understood. By adapting methods from soft matter physics, we dissect bacterial social behavior at the single cell level for several prototypical bacterial species, including Pseudomonas aeruginosa and Vibrio cholerae.

  13. Bacterial intermediate filaments

    DEFF Research Database (Denmark)

    Charbon, Godefroid; Cabeen, M.; Jacobs-Wagner, C.

    2009-01-01

    Crescentin, which is the founding member of a rapidly growing family of bacterial cytoskeletal proteins, was previously proposed to resemble eukaryotic intermediate filament (IF) proteins based on structural prediction and in vitro polymerization properties. Here, we demonstrate that crescentin...

  14. Bacterial Meningitis in Infants

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2004-04-01

    Full Text Available A retrospective study of 80 infantile patients (ages 30-365 days; 47 male, 33 female with culture-proven bacterial meningitis seen over a 16 year period (1986-2001 is reported from Taiwan.

  15. 8-Bromo-cyclic inosine diphosphoribose: towards a selective cyclic ADP-ribose agonist

    Science.gov (United States)

    Kirchberger, Tanja; Moreau, Christelle; Wagner, Gerd K.; Fliegert, Ralf; Siebrands, Cornelia C.; Nebel, Merle; Schmid, Frederike; Harneit, Angelika; Odoardi, Francesca; Flügel, Alexander; Potter, Barry V. L.; Guse, Andreas H.

    2009-01-01

    cADPR (cyclic ADP-ribose) is a universal Ca2+ mobilizing second messenger. In T-cells cADPR is involved in sustained Ca2+ release and also in Ca2+ entry. Potential mechanisms for the latter include either capacitative Ca2+ entry, secondary to store depletion by cADPR, or direct activation of the non-selective cation channel TRPM2 (transient receptor potential cation channel, subfamily melastatin, member 2). Here we characterize the molecular target of the newly-described membrane-permeant cADPR agonist 8-Br-N1-cIDPR (8-bromo-cyclic IDP-ribose). 8-Br-N1-cIDPR evoked Ca2+ signalling in the human T-lymphoma cell line Jurkat and in primary rat T-lymphocytes. Ca2+ signalling induced by 8-Br-N1-cIDPR consisted of Ca2+ release and Ca2+ entry. Whereas Ca2+ release was sensitive to both the RyR (ryanodine receptor) blocker RuRed (Ruthenium Red) and the cADPR antagonist 8-Br-cADPR (8-bromo-cyclic ADP-ribose), Ca2+ entry was inhibited by the Ca2+ entry blockers Gd3+ (gadolinium ion) and SKF-96365, as well as by 8-Br-cADPR. To unravel a potential role for TRPM2 in sustained Ca2+ entry evoked by 8-Br-N1-cIDPR, TRPM2 was overexpressed in HEK (human embryonic kidney)-293 cells. However, though activation by H2O2 was enhanced dramatically in those cells, Ca2+ signalling induced by 8-Br-N1-cIDPR was almost unaffected. Similarly, direct analysis of TRPM2 currents did not reveal activation or co-activation of TRPM2 by 8-Br-N1-cIDPR. In summary, the sensitivity to the Ca2+ entry blockers Gd3+ and SKF-96365 is in favour of the concept of capacitative Ca2+ entry, secondary to store depletion by 8-Br-N1-cIDPR. Taken together, 8-Br-N1-cIDPR appears to be the first cADPR agonist affecting Ca2+ release and secondary Ca2+ entry, but without effect on TRPM2. PMID:19492987

  16. Bacterial Cyanuric Acid Hydrolase for Water Treatment.

    Science.gov (United States)

    Yeom, Sujin; Mutlu, Baris R; Aksan, Alptekin; Wackett, Lawrence P

    2015-10-01

    Di- and trichloroisocyanuric acids are widely used as water disinfection agents, but cyanuric acid accumulates with repeated additions and must be removed to maintain free hypochlorite for disinfection. This study describes the development of methods for using a cyanuric acid-degrading enzyme contained within nonliving cells that were encapsulated within a porous silica matrix. Initially, three different bacterial cyanuric acid hydrolases were compared: TrzD from Acidovorax citrulli strain 12227, AtzD from Pseudomonas sp. strain ADP, and CAH from Moorella thermoacetica ATCC 39073. Each enzyme was expressed recombinantly in Escherichia coli and tested for cyanuric acid hydrolase activity using freely suspended or encapsulated cell formats. Cyanuric acid hydrolase activities differed by only a 2-fold range when comparing across the different enzymes with a given format. A practical water filtration system is most likely to be used with nonviable cells, and all cells were rendered nonviable by heat treatment at 70°C for 1 h. Only the CAH enzyme from the thermophile M. thermoacetica retained significant activity under those conditions, and so it was tested in a flowthrough system simulating a bioreactive pool filter. Starting with a cyanuric acid concentration of 10,000 μM, more than 70% of the cyanuric acid was degraded in 24 h, it was completely removed in 72 h, and a respike of 10,000 μM cyanuric acid a week later showed identical biodegradation kinetics. An experiment conducted with water obtained from municipal swimming pools showed the efficacy of the process, although cyanuric acid degradation rates decreased by 50% in the presence of 4.5 ppm hypochlorite. In total, these experiments demonstrated significant robustness of cyanuric acid hydrolase and the silica bead materials in remediation.

  17. CRED Acoustic Doppler Profiler (ADP); NWHI, MID; Long: -177.42181, Lat: 28.21826 (WGS84); Sensor Depth: 1.83m; Data Range: 20080926-20090321.

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Data from Coral Reef Ecosystem Division (CRED), NOAA Pacific Island Fisheries Science Center Acoustic Doppler Profilers (ADP) provide a time series of water current...

  18. Effect of Ad-p16 Combined with CDDP or As2O3 on Human Bladder Cancer Cells

    Institute of Scientific and Technical Information of China (English)

    朱朝辉; 邢诗安; 林晨

    2003-01-01

    Summary: To evaluate the therapeutic efficiency of combined use of p16-expressing adenovirus and chemotherapeutic agents CDDP or As2O3 on human bladder cancer cell line E J, the human bladder cancer cell line EJ were transfected with adenovirus-mediated p16 gene (Ad-p16), with administration of cisplatin (CDDP) or arsenic trioxide (As2O3). The cell growth, morphological changes, cell cycle, apoptosis and molecular changes were measured using cell counting, reverse microscopy, flow cytometry, cloning formation, immunocytochemical assays and in vivo therapy experiments to evaluate the therapeutic efficacy of such combined regimen. Ad-p16 transfer and CDDP or As2O3 administration to EJ cells could exert substantially stronger therapeutic effects than the single agent treatment. Especially in in vivo experiments, combined administration of p16 and CDDP or As2O3 induced almost tumor diminish compared to the partial tumor diminish induced by single agent. Moreover,delivery of Ad-p16, or administration of minimal-dose CDDP or As2O3 or combined regimen could induce massive apoptosis of EJ cell. Cell cycle analysis demonstrated that administration of CDDP or As2O3 remarkably arrested EJ cell in G1 prior to apoptotic cell death. When treated with combined regimen, cells were arrested in G1 to a greater extent prior to apoptotic cell death. It is concluded that after introduction into EJ cell, Ad-p16 shows enhanced therapeutic efficacy for EJ cell when used in combination with CDDP or As2O3.

  19. Phylogenetic analysis of the thylakoid ATP/ADP carrier reveals new insights into its function restricted to green plants

    OpenAIRE

    Cornelia eSpetea; Pfeil, Bernard E.; Benoit eSchoefs

    2012-01-01

    ATP is the common energy currency of cellular metabolism in all living organisms. Most of them synthesize ATP in the cytosol or on the mitochondrial inner membrane, whereas land plants, algae and cyanobacteria also produce it on the thylakoid membrane during the light-dependent reactions of photosynthesis. From the site of synthesis, ATP is transported to the site of utilization via intracellular membranes transporters. One major type of ATP transporter is represented by the mitochondrial ADP...

  20. Role of poly(ADP-ribose) polymerase 1 in DNA methylation changes induced by hydroquinone in human bronchial epithelial cell

    Institute of Scientific and Technical Information of China (English)

    沙炎

    2014-01-01

    Objective To investigate the DNA methylation changes induced by hydroquinone(HQ)in human bronchial epithelial cells and to explore the role of poly(ADP-ribose)polymerase-1(PARP-1)in this process.Methods Human bronchial epithelial 16HBE cells and PARP-1-deficient 16HBE cells(16HBE-shPARP-1 cells)were exposed to HQ(10,20,40,60,and 80μmol/L)for 48

  1. The ADP-ribose polymerase Tankyrase regulates adult intestinal stem cell proliferation during homeostasis in Drosophila.

    Science.gov (United States)

    Wang, Zhenghan; Tian, Ai; Benchabane, Hassina; Tacchelly-Benites, Ofelia; Yang, Eungi; Nojima, Hisashi; Ahmed, Yashi

    2016-05-15

    Wnt/β-catenin signaling controls intestinal stem cell (ISC) proliferation, and is aberrantly activated in colorectal cancer. Inhibitors of the ADP-ribose polymerase Tankyrase (Tnks) have become lead therapeutic candidates for Wnt-driven cancers, following the recent discovery that Tnks targets Axin, a negative regulator of Wnt signaling, for proteolysis. Initial reports indicated that Tnks is important for Wnt pathway activation in cultured human cell lines. However, the requirement for Tnks in physiological settings has been less clear, as subsequent studies in mice, fish and flies suggested that Tnks was either entirely dispensable for Wnt-dependent processes in vivo, or alternatively, had tissue-specific roles. Here, using null alleles, we demonstrate that the regulation of Axin by the highly conserved Drosophila Tnks homolog is essential for the control of ISC proliferation. Furthermore, in the adult intestine, where activity of the Wingless pathway is graded and peaks at each compartmental boundary, Tnks is dispensable for signaling in regions where pathway activity is high, but essential where pathway activity is relatively low. Finally, as observed previously for Wingless pathway components, Tnks activity in absorptive enterocytes controls the proliferation of neighboring ISCs non-autonomously by regulating JAK/STAT signaling. These findings reveal the requirement for Tnks in the control of ISC proliferation and suggest an essential role in the amplification of Wnt signaling, with relevance for development, homeostasis and cancer. PMID:27190037

  2. Molecular characterization of multiple cDNA clones for ADP-glucose pyrophosphorylase from Arabidopsis thaliana.

    Science.gov (United States)

    Villand, P; Olsen, O A; Kleczkowski, L A

    1993-12-01

    PCR amplification of cDNA prepared from poly(A)+ RNA from aerial parts of Arabidopsis thaliana, using degenerate nucleotide primers based on conserved regions between the large and small subunits of ADP-glucose pyrophosphorylase (AGP), yielded four different cDNAs of ca. 550 nucleotides each. Based on derived amino acid sequences, the identities between the clones varied from 49 to 69%. Sequence comparison to previously published cDNAs for AGP from various species and tissues has revealed that three of the amplified cDNAs (ApL1, ApL2 and ApL3) correspond to the large subunit of AGP, and one cDNA (ApS) encodes the small subunit of AGP. Both ApL1 and ApS were subsequently found to be present in a cDNA library made from Arabidopsis leaves. All four PCR products are encoded by single genes, as found by genomic Southern analysis. PMID:8292792

  3. Docking study and binding free energy calculation of poly (ADP-ribose) polymerase inhibitors.

    Science.gov (United States)

    Ohno, Kazuki; Mitsui, Takashi; Tanida, Yoshiaki; Matsuura, Azuma; Fujitani, Hideaki; Niimi, Tatsuya; Orita, Masaya

    2011-02-01

    Recently, the massively parallel computation of absolute binding free energy with a well-equilibrated system (MP-CAFEE) has been developed. The present study aimed to determine whether the MP-CAFEE method is useful for drug discovery research. In the drug discovery process, it is important for computational chemists to predict the binding affinity accurately without detailed structural information for protein/ligand complex. We investigated the absolute binding free energies for Poly (ADP-ribose) polymerase-1 (PARP-1)/inhibitor complexes, using the MP-CAFEE method. Although each docking model was used as an input structure, it was found that the absolute binding free energies calculated by MP-CAFEE are well consistent with the experimental ones. The accuracy of this method is much higher than that using molecular mechanics Poisson-Boltzmann/surface area (MM/PBSA). Although the simulation time is quite extensive, the reliable predictor of binding free energies would be a useful tool for drug discovery projects. PMID:20480380

  4. Inhibition of poly(ADP-ribose)polymerase-1 and DNA repair by uranium.

    Science.gov (United States)

    Cooper, Karen L; Dashner, Erica J; Tsosie, Ranalda; Cho, Young Mi; Lewis, Johnnye; Hudson, Laurie G

    2016-01-15

    Uranium has radiological and non-radiological effects within biological systems and there is increasing evidence for genotoxic and carcinogenic properties attributable to uranium through its heavy metal properties. In this study, we report that low concentrations of uranium (as uranyl acetate; uranium exacerbates DNA damage and cytotoxicity induced by hydrogen peroxide, suggesting that uranium may inhibit DNA repair processes. Concentrations of uranyl acetate in the low micromolar range inhibited the zinc finger DNA repair protein poly(ADP-ribose) polymerase (PARP)-1 and caused zinc loss from PARP-1 protein. Uranyl acetate exposure also led to zinc loss from the zinc finger DNA repair proteins Xeroderma Pigmentosum, Complementation Group A (XPA) and aprataxin (APTX). In keeping with the observed inhibition of zinc finger function of DNA repair proteins, exposure to uranyl acetate enhanced retention of induced DNA damage. Co-incubation of uranyl acetate with zinc largely overcame the impact of uranium on PARP-1 activity and DNA damage. These findings present evidence that low concentrations of uranium can inhibit DNA repair through disruption of zinc finger domains of specific target DNA repair proteins. This may provide a mechanistic basis to account for the published observations that uranium exposure is associated with DNA repair deficiency in exposed human populations.

  5. Poly (ADP-Ribose Polymerase Mediates Diabetes-Induced Retinal Neuropathy

    Directory of Open Access Journals (Sweden)

    Ghulam Mohammad

    2013-01-01

    Full Text Available Retinal neuropathy is an early event in the development of diabetic retinopathy. One of the potential enzymes that are activated by oxidative stress in the diabetic retina is poly (ADP-ribose polymerase (PARP. We investigated the effect of the PARP inhibitor 1,5-isoquinolinediol on the expression of the neurodegeneration mediators and markers in the retinas of diabetic rats. After two weeks of streptozotocin-induced diabetes, rats were treated with 1,5-isoquinolinediol (3 mg/kg/day. After 4 weeks of diabetes, the retinas were harvested and the levels of reactive oxygen species (ROS were determined fluorometrically and the expressions of PARP, phosporylated-ERK1/2, BDNF, synaptophysin, glutamine synthetase (GS, and caspase-3 were determined by Western blot analysis. Retinal levels of ROS, PARP-1/2, phosphorylated ERK1/2, and cleaved caspase-3 were significantly increased, whereas the expressions of BDNF synaptophysin and GS were significantly decreased in the retinas of diabetic rats, compared to nondiabetic rats. Administration of 1,5-isoquinolinediol did not affect the metabolic status of the diabetic rats, but it significantly attenuated diabetes-induced upregulation of PARP, ROS, ERK1/2 phosphorylation, and cleaved caspase-3 and downregulation of BDNF, synaptophysin, and GS. These findings suggest a beneficial effect of the PARP inhibitor in increasing neurotrophic support and ameliorating early retinal neuropathy induced by diabetes.

  6. Sucrose regulation of ADP-glucose pyrophosphorylase subunit genes transcript levels in leaves and fruits

    Science.gov (United States)

    Li, Xiangyang; Xing, Jinpeng; Gianfagna, Thomas J.; Janes, Harry W.

    2002-01-01

    ADP-glucose pyrophosphorylase (AGPase, EC2.7.7.27) is a key regulatory enzyme in starch biosynthesis. The enzyme is a heterotetramer with two S and two B subunits. In tomato, there are three multiple forms of the S subunit gene. Agp S1, S2 and B are highly expressed in fruit from 10 to 25 days after anthesis. Agp S3 is only weakly expressed in fruit. Sucrose significantly elevates expression of Agp S1, S2 and B in both leaves and fruits. Agp S1 exhibits the highest degree of regulation by sucrose. In fact, sucrose may be required for Agp S1 expression. For excised leaves incubated in water, no transcripts for Agp S1 could be detected in the absence of sucrose, whereas it took up to 16 h in water before transcripts were no longer detectable for Agp S2 and B. Neither Agp S3 nor the tubulin gene is affected by sucrose, demonstrating that this response is specifically regulated by a carbohydrate metabolic signal, and is not due to a general increase in metabolism caused by sucrose treatment. Truncated versions of the promoter for Agp S1 indicate that a specific region 1.3-3.0 kb upstream from the transcription site is responsible for sucrose sensitivity. This region of the S1 promoter contains several cis-acting elements present in the promoters of other genes that are also regulated by sucrose. c2002 Elsevier Science Ireland Ltd. All rights reserved.

  7. Poly (ADP-ribose polymerase 1 protein expression in normal and neoplastic prostatic tissue

    Directory of Open Access Journals (Sweden)

    M. Salemi

    2013-04-01

    Full Text Available A genetic background has been implicated in the development of prostate cancer. Protein microarrays have enabled the identification of proteins, some of which associated with apoptosis, that may play a role in the development of such a tumor. Inhibition of apoptosis is a co-factor that contributes to the onset and progression of prostate cancer, though the molecular mechanisms are not entirely understood. Poly (ADP-ribose polymerase 1 (PARP-1 gene is required for translocation of the apoptosis-inducing factor (AIF from the mitochondria to the nucleus. Hence, it is involved in programmed cell death. Different PARP-1 gene expression has been observed in various tumors such as glioblastoma, lung, ovarian, endometrial, and skin cancers. We evaluated the expression of PARP-1 protein in prostatic cancer and normal prostate tissues by immunohistochemistry in 40 men with prostate cancer and in 37 normal men. Positive nuclear PARP-1 staining was found in all samples (normal prostate and prostate cancer tissues. No cytoplasmic staining was observed in any sample. PARP-1-positive cells resulted significantly higher in patients with prostate carcinoma compared with controls (P<0.001. PARP-1 over-expression in prostate cancer tissue compared with normal prostate suggests a greater activity of PARP-1 in these tumors. These findings suggest that PARP-1 expression in prostate cancer is an attempt to trigger apoptosis in this type of tumor similarly to what reported in other cancers.

  8. Green Tea Polyphenols Control Dysregulated Glutamate Dehydrogenase in Transgenic Mice by Hijacking the ADP Activation Site

    Energy Technology Data Exchange (ETDEWEB)

    Li, Changhong; Li, Ming; Chen, Pan; Narayan, Srinivas; Matschinsky, Franz M.; Bennett, Michael J.; Stanley, Charles A.; Smith, Thomas J. (CH-PA); (UPENN); (Danforth)

    2012-05-09

    Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of L-glutamate and, in animals, is extensively regulated by a number of metabolites. Gain of function mutations in GDH that abrogate GTP inhibition cause the hyperinsulinism/hyperammonemia syndrome (HHS), resulting in increased pancreatic {beta}-cell responsiveness to leucine and susceptibility to hypoglycemia following high protein meals. We have previously shown that two of the polyphenols from green tea (epigallocatechin gallate (EGCG) and epicatechin gallate (ECG)) inhibit GDH in vitro and that EGCG blocks GDH-mediated insulin secretion in wild type rat islets. Using structural and site-directed mutagenesis studies, we demonstrate that ECG binds to the same site as the allosteric regulator, ADP. Perifusion assays using pancreatic islets from transgenic mice expressing a human HHS form of GDH demonstrate that the hyperresponse to glutamine caused by dysregulated GDH is blocked by the addition of EGCG. As observed in HHS patients, these transgenic mice are hypersensitive to amino acid feeding, and this is abrogated by oral administration of EGCG prior to challenge. Finally, the low basal blood glucose level in the HHS mouse model is improved upon chronic administration of EGCG. These results suggest that this common natural product or some derivative thereof may prove useful in controlling this genetic disorder. Of broader clinical implication is that other groups have shown that restriction of glutamine catabolism via these GDH inhibitors can be useful in treating various tumors. This HHS transgenic mouse model offers a highly useful means to test these agents in vivo.

  9. PKCα and HMGB1 antagonistically control hydrogen peroxide-induced poly-ADP-ribose formation

    Science.gov (United States)

    Andersson, Anneli; Bluwstein, Andrej; Kumar, Nitin; Teloni, Federico; Traenkle, Jens; Baudis, Michael; Altmeyer, Matthias; Hottiger, Michael O.

    2016-01-01

    Harmful oxidation of proteins, lipids and nucleic acids is observed when reactive oxygen species (ROS) are produced excessively and/or the antioxidant capacity is reduced, causing ‘oxidative stress’. Nuclear poly-ADP-ribose (PAR) formation is thought to be induced in response to oxidative DNA damage and to promote cell death under sustained oxidative stress conditions. However, what exactly triggers PAR induction in response to oxidative stress is incompletely understood. Using reverse phase protein array (RPPA) and in-depth analysis of key stress signaling components, we observed that PAR formation induced by H2O2 was mediated by the PLC/IP3R/Ca2+/PKCα signaling axis. Mechanistically, H2O2-induced PAR formation correlated with Ca2+-dependent DNA damage, which, however, was PKCα-independent. In contrast, PAR formation was completely lost upon knockdown of PKCα, suggesting that DNA damage alone was not sufficient for inducing PAR formation, but required a PKCα-dependent process. Intriguingly, the loss of PAR formation observed upon PKCα depletion was overcome when the chromatin structure-modifying protein HMGB1 was co-depleted with PKCα, suggesting that activation and nuclear translocation of PKCα releases the inhibitory effect of HMGB1 on PAR formation. Together, these results identify PKCα and HMGB1 as important co-regulators involved in H2O2-induced PAR formation, a finding that may have important relevance for oxidative stress-associated pathophysiological conditions. PMID:27198223

  10. Green tea polyphenols control dysregulated glutamate dehydrogenase in transgenic mice by hijacking the ADP activation site.

    Science.gov (United States)

    Li, Changhong; Li, Ming; Chen, Pan; Narayan, Srinivas; Matschinsky, Franz M; Bennett, Michael J; Stanley, Charles A; Smith, Thomas J

    2011-09-30

    Glutamate dehydrogenase (GDH) catalyzes the oxidative deamination of L-glutamate and, in animals, is extensively regulated by a number of metabolites. Gain of function mutations in GDH that abrogate GTP inhibition cause the hyperinsulinism/hyperammonemia syndrome (HHS), resulting in increased pancreatic β-cell responsiveness to leucine and susceptibility to hypoglycemia following high protein meals. We have previously shown that two of the polyphenols from green tea (epigallocatechin gallate (EGCG) and epicatechin gallate (ECG)) inhibit GDH in vitro and that EGCG blocks GDH-mediated insulin secretion in wild type rat islets. Using structural and site-directed mutagenesis studies, we demonstrate that ECG binds to the same site as the allosteric regulator, ADP. Perifusion assays using pancreatic islets from transgenic mice expressing a human HHS form of GDH demonstrate that the hyperresponse to glutamine caused by dysregulated GDH is blocked by the addition of EGCG. As observed in HHS patients, these transgenic mice are hypersensitive to amino acid feeding, and this is abrogated by oral administration of EGCG prior to challenge. Finally, the low basal blood glucose level in the HHS mouse model is improved upon chronic administration of EGCG. These results suggest that this common natural product or some derivative thereof may prove useful in controlling this genetic disorder. Of broader clinical implication is that other groups have shown that restriction of glutamine catabolism via these GDH inhibitors can be useful in treating various tumors. This HHS transgenic mouse model offers a highly useful means to test these agents in vivo.

  11. Differential Role of Poly(ADP-ribose polymerase in D. discoideum growth and development

    Directory of Open Access Journals (Sweden)

    Begum Rasheedunnisa

    2011-03-01

    Full Text Available Abstract Background Poly(ADP-ribose polymerase is evolutionarily conserved as a responder to various forms of stress. Though PARP's role in cell death is well addressed, its role in development and multicellularity is still an enigma. We have previously reported the role of PARP in oxidative stress induced delayed development of D. discoideum. Results In the current study we highlight the involvement of PARP during D. discoideum development. Oxidative stress affects expression of aca and cAR1 thus affecting aggregation. Although parp expression is not affected during oxidative stress but it is involved during normal development as confirmed by our PARP down-regulation studies. Constitutive PARP down-regulation resulted in blocked development while no effect was observed on D. discoideum growth. Interestingly, stage specific PARP down-regulation arrested development at the slug stage. Conclusion These results emphasize that PARP is essential for complex differentiation and its function may be linked to multicellularity. This is the first report where the involvement of PARP during normal multicellular development in D. discoideum, an ancient eukaryote, is established which could be of evolutionary significance. Thus our study adds one more role to the multitasking function of PARP.

  12. Metabotropic glutamate receptor activation and intracellular cyclic ADP-ribose release Ca2+ from the same store in cultured DRG neurones.

    Science.gov (United States)

    Pollock, J; Crawford, J H; Wootton, J F; Seabrook, G R; Scott, R H

    1999-01-01

    The whole cell patch clamp technique has been used to record Ca(2+)-activated cation and chloride conductances evoked by release of Ca2+ from intracellular stores of cultured neonatal dorsal root ganglion neurones. The aim of this study was to investigate metabotropic glutamate receptor (mGluR) mechanisms and evaluate a possible role for cyclic ADP-ribose as an intracellular signalling molecule. Glutamate and the metabotropic glutamate receptor agonist (1S, 3R)-ACPD-evoked transient depolarizations, Ca(2+)-activated inward currents and rises in intracellular Ca2+. The (1S, 3R)-ACPD-activated currents were insensitive to InsP3 signalling inhibitors, heparin and pentosan polysulphate. Intracellular application of ryanodine alone activated currents in this study and proved a difficult tool to use as a potential inhibitor of cyclic ADP-ribose-mediated responses. However, intracellular dantrolene did attenuate both (1S, 3R)-ACPD and cyclic ADP-ribose responses. Intracellular photo-release of cGMP and cyclic ADP-ribose mimicked the responses to mGluR receptor activation. Intracellular application of nicotinamide and W7 inhibited the responses to photo-released cGMP but did not prevent responses to mGluR activation. The cyclic ADP-ribose receptor antagonist 8-amino cyclic ADP-ribose attenuated responses to (1S, 3R)-ACPD, cGMP and cyclic ADP-ribose, but some Ca(2+)-activated inward currents were still observed in the presence of this antagonist. In conclusion, mGluR receptor activation, cGMP and cyclic ADP-ribose release Ca2+ from intracellular stores. Some evidence suggests that pharmacologically related pathways are involved. PMID:10598278

  13. 2´,3´-Dialdehyde of ATP, ADP, and adenosine inhibit HIV-1 reverse transcriptase and HIV-1 replication.

    Science.gov (United States)

    Schachter, Julieta; Valadao, Ana Luiza Chaves; Aguiar, Renato Santana; Barreto-de-Souza, Victor; Rossi, Atila Duque; Arantes, Pablo Ricardo; Verli, Hugo; Quintana, Paula Gabriela; Heise, Norton; Tanuri, Amilcar; Bou-Habib, Dumith Chequer; Persechini, Pedro Muanis

    2014-01-01

    The 2´3´-dialdehyde of ATP or oxidized ATP (oATP) is a compound known for specifically making covalent bonds with the nucleotide-binding site of several ATP-binding enzymes and receptors. We investigated the effects of oATP and other oxidized purines on HIV-1 infection and we found that this compound inhibits HIV-1 and SIV infection by blocking early steps of virus replication. oATP, oxidized ADP (oADP), and oxidized Adenosine (oADO) impact the natural activity of endogenous reverse transcriptase enzyme (RT) in cell free virus particles and are able to inhibit viral replication in different cell types when added to the cell cultures either before or after infection. We used UFLC-UV to show that both oADO and oATP can be detected in the cell after being added in the extracellular medium. oATP also suppresses RT activity and replication of the HIV-1 resistant variants M184V and T215Y. We conclude that oATP, oADP and oADO display anti HIV-1 activity that is at in least in part due to inhibitory activity on HIV-1 RT.

  14. Inhibitory effects and mechanisms of high molecular-weight phlorotannins from Sargassum thunbergii on ADP-induced platelet aggregation

    Institute of Scientific and Technical Information of China (English)

    WEI Yuxi; WANG Changyun; LI Jing; GUO Qi; QI Hongtao

    2009-01-01

    We evaluated the effects of high molecular-weight phlorotannins from Sargassum thunbergii (STP) on ADP-induced platelet aggregation and arachidonic acid (AA) metabolism in New Zealand white rabbits and Wistar rats. The inhibition of STP on platelet aggregation was investigated using a turbidimetric method, and the levels of the terminal products of AA metabolism were measured using the corresponding kits for maleic dialdehyde (MDA), thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-keto-PGF1α) by colorimetry and radioimmunoassay, as appropriate. We found that STP could inhibit ADP-induced platelet aggregation, and the inhibitory ratio was 91.50% at the STP concentration of 4.0 mg/mL. Furthermore, STP markedly affected AA metabolism by decreasing the synthesis of MDA (P<0.01) and increasing the synthesis of 6-keto-PGF1α, thus changing the plasma TXB2/6-keto-PGF1α balance when the platelets were activated (P<0.01). Therefore, STP altered AA metabolism and these findings partly revealed the molecular mechanism by which STP inhibits ADP-induced platelet aggregation.

  15. A study of the expression of p53 in posttransfection cells with rAdp53 gene and inhibitory activity in vitro

    Institute of Scientific and Technical Information of China (English)

    Jianhua Wang; Zongzheng Ji; Xiaoqiang Wang

    2007-01-01

    Objective: To investigate the inhibitory effect and IC50 (50% inhibiting concentration) of the recombinant adenoviral p53 gene (rAdp53) in colorectal cancer cells in vitro and to guide clinical practice. Methods: We evaluated the efficiency (IC50)of the rAdp53 and six kinds of anti-cancer drugs(5-fluorouracil, tegafur, mitomycin c, cisplatin, oxaliplatin, paclitaxel) in human colorectal cancer cell line-174 through the cell culture and MTT chemosensitivity assay to make sure the anti-cancer capability of rAdp53.Expression of p53 protein in transfection cells of colorectal cancer line-174 with rAdp53 was evaluated by immunohistochemical staining. Results: The rAdp53 is a dose- and time-dependent anti-cancer drug, its IC5o is 5.73×1011 VP/ml, but its effect was not obvious when compared with other anti-cancer drugs. In control group, the immunohistochemistry stain was negative. However,rAd-p53 of five different concentrations were all positive in infected colorectal cancer cells with rAd-p53 and the earliest positive result would present 24 hours after infection. Conclusion: The rAdp53 has good anti-cancer efficacy is colorectal cancer cell line174 in vitro. But its anti-cancer efficacy was less than those of the classical chemical medicine mitomycin c, 5-fluorouracil and cisplatin etc., when it was used alone.

  16. The bacterial lipocalins.

    Science.gov (United States)

    Bishop, R E

    2000-10-18

    The lipocalins were once regarded as a eukaryotic protein family, but new members have been recently discovered in bacteria. The first bacterial lipocalin (Blc) was identified in Escherichia coli as an outer membrane lipoprotein expressed under conditions of environmental stress. Blc is distinguished from most lipocalins by the absence of intramolecular disulfide bonds, but the presence of a membrane anchor is shared with two of its closest homologues, apolipoprotein D and lazarillo. Several common features of the membrane-anchored lipocalins suggest that each may play an important role in membrane biogenesis and repair. Additionally, Blc proteins are implicated in the dissemination of antibiotic resistance genes and in the activation of immunity. Recent genome sequencing efforts reveal the existence of at least 20 bacterial lipocalins. The lipocalins appear to have originated in Gram-negative bacteria and were probably transferred horizontally to eukaryotes from the endosymbiotic alpha-proteobacterial ancestor of the mitochondrion. The genome sequences also reveal that some bacterial lipocalins exhibit disulfide bonds and alternative modes of subcellular localization, which include targeting to the periplasmic space, the cytoplasmic membrane, and the cytosol. The relationships between bacterial lipocalin structure and function further illuminate the common biochemistry of bacterial and eukaryotic cells.

  17. Diagnosis of bacterial vaginosis

    Directory of Open Access Journals (Sweden)

    Đukić Slobodanka

    2013-01-01

    Full Text Available Bacterial vaginosis is a common, complex clinical syndrome characterized by alterations in the normal vaginal flora. When symptomatic, it is associated with a malodorous vaginal discharge and on occasion vaginal burning or itching. Under normal conditions, lactobacilli constitute 95% of the bacteria in the vagina. Bacterial vaginosis is associated with severe reduction or absence of the normal H2O2­producing lactobacilli and overgrowth of anaerobic bacteria and Gardnerella vaginalis, Atopobium vaginae, Mycoplasma hominis and Mobiluncus species. Most types of infectious disease are diagnosed by culture, by isolating an antigen or RNA/DNA from the microbe, or by serodiagnosis to determine the presence of antibodies to the microbe. Therefore, demonstration of the presence of an infectious agent is often a necessary criterion for the diagnosis of the disease. This is not the case for bacterial vaginosis, since the ultimate cause of the disease is not yet known. There are a variety of methods for the diagnosis of bacterial vaginosis but no method can at present be regarded as the best. Diagnosing bacterial vaginosis has long been based on the clinical criteria of Amsel, whereby three of four defined criteria must be satisfied. Nugent’s scoring system has been further developed and includes validation of the categories of observable bacteria structures. Up­to­date molecular tests are introduced, and better understanding of vaginal microbiome, a clear definition for bacterial vaginosis, and short­term and long­term fluctuations in vaginal microflora will help to better define molecular tests within the broader clinical context.

  18. Bacterial glycosyltransferase toxins.

    Science.gov (United States)

    Jank, Thomas; Belyi, Yury; Aktories, Klaus

    2015-12-01

    Mono-glycosylation of host proteins is a common mechanism by which bacterial protein toxins manipulate cellular functions of eukaryotic target host cells. Prototypic for this group of glycosyltransferase toxins are Clostridium difficile toxins A and B, which modify guanine nucleotide-binding proteins of the Rho family. However, toxin-induced glycosylation is not restricted to the Clostridia. Various types of bacterial pathogens including Escherichia coli, Yersinia, Photorhabdus and Legionella species produce glycosyltransferase toxins. Recent studies discovered novel unexpected variations in host protein targets and amino acid acceptors of toxin-catalysed glycosylation. These findings open new perspectives in toxin as well as in carbohydrate research.

  19. Sensitization of prostate cancer to ionizing radiation by targeting poly(ADP-robose) polymerase: preclinical studies

    International Nuclear Information System (INIS)

    Poly(ADP-ribose) polymerase (PARP) is a DNA-binding enzyme which plays important roles in the maintenance of genome stability, immediate cellular responses to DNA damage, and apoptosis. A DNA-binding domain of PARP (PARP-DBD) acts as a dominant-negative mutant by binding to DNA strand breaks irreversibly and sensitizing mammalian cells to DNA-damaging agents (1, 2). To direct the expression of human PARP-DBD to prostate we developed recombinant plasmids expressing the PARP-DBD under the control of the 5'-flanking sequences of the human prostate-specific antigen (PSA) gene. In vitro studies revealed that PSA promoter driven expression of the PARP-DBD showed prostate tissue specificity and androgen responsiveness and sensitized LNCaP cells to DNA-damaging agents, such as ionizing radiation and etoposide (3). To assess the efficiency of this strategy in vivo, we developed a cationic liposome-mediated gene delivery of PARP-DBD plasmid in tumor xenografts of PSA producing and androgen sensitive prostate cancer cells (LNCaP and 22Rv1). Tumor bearing mice were treated with intratumoral liposome-complexed PARP-DBD (LE-PARP-DBD), ionizing radiation (IR) or a combination of LE-PARP-DBD and IR. Control groups received blank liposomes or were left untreated. Administration of LE-PARP-DBD resulted in expression of dominant-negative mutant of PARP in tumor cells and enhanced radiation-induced inhibition of tumor growth. These results provide a proof-of- principle for a novel therapeutic strategy to control prostate cancer. The study was supported in part by grants from the U.S. Army Medical Research and Development Command DAMD 17-00-1-0019 and DAMD 17-00-1-0276 (to V.S.). (1) J.Biol.Chem., 265:18721-18724, 1990; (2) Cancer Research, 58: 3495-3498, 1998; (3) Cancer Research, 62: 6879-6883, 2002

  20. Poly(Adp-ribose) synthetase inhibition prevents lipopolysaccharide-induced peroxynitrite mediated damage in diaphragm.

    Science.gov (United States)

    Ozdülger, Ali; Cinel, Ismail; Unlü, Ali; Cinel, Leyla; Mavioglu, Ilhan; Tamer, Lülüfer; Atik, Ugur; Oral, Ugur

    2002-07-01

    Although the precise mechanism by which sepsis causes impairment of respiratory muscle contractility has not been fully elucidated, oxygen-derived free radicals are thought to play an important role. In our experimental study, the effects of poly(ADP-ribose) synthetase (PARS) inhibition on the diaphragmatic Ca(2+)-ATPase, malondialdehyde (MDA), and 3-nitrotyrosine (3-NT) levels and additionally histopathology of the diaphragm in lipopolysaccharide (LPS)-induced endotoxemia are investigated.Thirty-two male Wistar rats, weighing between 180-200 g were randomly divided into four groups. The first group (control; n=8) received saline solution and the second (LPS group; n=8) 10 mgkg(-1) LPS i.p. 3-Aminobenzamide (3-AB) as a PARS inhibitor; was given to the third group (C+3-AB, n=8) 20 min before administration of saline solution while the fourth group (LPS+3-AB, n=8) received 3-AB 20 min before LPS injection. Six hours later, under ketamin/xylasine anesthesia diapraghmatic specimens were obtained and the rats were decapitated. Diaphragmatic specimens were divided into four parts, three for biochemical analyses and one for histopathologic assessment. In the LPS group, tissue Ca(2+)-ATPase levels were found to be decreased and tissue MDA and 3-NT levels were found to be increased (P<0.05). In the LPS+3-AB group, 3-AB pretreatment inhibited the increase in MDA and 3-NT levels and Ca(2+)-ATPase activity remained similar to those in the control group (P<0.05). Histopathologic examination of diaphragm showed edema between muscle fibers only in LPS group. PARS inhibition with 3-AB prevented not only lipid peroxidation but also the decrease of Ca(2+)-ATPase activity in endotoxemia. These results highlights the importance of nitric oxide (NO)-peroxynitrite (ONOO(-))-PARS pathway in preventing free radical mediated injury. PARS inhibitors should further be investigated as a new thearapetic alternative in sepsis treatment.