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Sample records for telomere-specific nucleosome mobility

  1. Influence of histone tails and H4 tail acetylations on nucleosome-nucleosome interactions.

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    Liu, Ying; Lu, Chenning; Yang, Ye; Fan, Yanping; Yang, Renliang; Liu, Chuan-Fa; Korolev, Nikolay; Nordenskiöld, Lars

    2011-12-16

    Nucleosome-nucleosome interaction plays a fundamental role in chromatin folding and self-association. The cation-induced condensation of nucleosome core particles (NCPs) displays properties similar to those of chromatin fibers, with important contributions from the N-terminal histone tails. We study the self-association induced by addition of cations [Mg(2+), Ca(2+), cobalt(III)hexammine(3+), spermidine(3+) and spermine(4)(+)] for NCPs reconstituted with wild-type unmodified histones and with globular tailless histones and for NCPs with the H4 histone tail having lysine (K) acetylations or lysine-to-glutamine mutations at positions K5, K8, K12 and K16. In addition, the histone construct with the single H4K16 acetylation was investigated. Acetylated histones were prepared by a semisynthetic native chemical ligation method. The aggregation behavior of NCPs shows a general cation-dependent behavior similar to that of the self-association of nucleosome arrays. Unlike nucleosome array self-association, NCP aggregation is sensitive to position and nature of the H4 tail modification. The tetra-acetylation in the H4 tail significantly weakens the nucleosome-nucleosome interaction, while the H4 K→Q tetra-mutation displays a more modest effect. The single H4K16 acetylation also weakens the self-association of NCPs, which reflects the specific role of H4K16 in the nucleosome-nucleosome stacking. Tailless NCPs can aggregate in the presence of oligocations, which indicates that attraction also occurs by tail-independent nucleosome-nucleosome stacking and DNA-DNA attraction in the presence of cations. The experimental data were compared with the results of coarse-grained computer modeling for NCP solutions with explicit presence of mobile ions. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. High-mobility group nucleosome-binding domain 2 protein inhibits the invasion of Klebsiella pneumoniae into mouse lungs in vivo.

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    Zheng, Shuang; Ren, Laibin; Li, Heng; Shen, Xiaofei; Yang, Xiaolong; Li, Na; Wang, Xinyuan; Guo, Xiaojuan; Wang, Xiaoying; Huang, Ning

    2015-07-01

    Since bacterial invasion into host cells is a critical step in the infection process and the predominance of multiple-antibiotic-resistant Klebsiella (K.) pneumoniae strains, using molecular agents to interfere with K. pneumoniae invasion is an attractive approach for the prevention of infection and suppress the immune inflammatory response. In previous studies by our group, high-mobility group nucleosome-binding domain 2 (HMGN2) protein was shown to exhibit anti-bacterial activity in vitro. The objective of the present study was to investigate the effects of HMGN2 protein on the invasion of K. pneumoniae 03183 in vivo. The results showed that pre-treatment with 128 µg/ml HMGN2 significantly reduced K. pneumoniae 03183 invasion into mouse lungs and increased the mRNA expression of CXCL1 and LCN2 within 2 h. Immunohistochemical staining showed that F-actin expression was significantly decreased, and fluorescence microscopy and western blot analysis further demonstrated that HMGN2 significantly blocked K. pneumoniae 03183-induced actin polymerization. These changes implied that HMGN2 may provide protection against K. pneumoniae 03183 infection in vivo.

  3. TRF2 controls telomeric nucleosome organization in a cell cycle phase-dependent manner.

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    Alessandra Galati

    Full Text Available Mammalian telomeres stabilize chromosome ends as a result of their assembly into a peculiar form of chromatin comprising a complex of non-histone proteins named shelterin. TRF2, one of the shelterin components, binds to the duplex part of telomeric DNA and is essential to fold the telomeric chromatin into a protective cap. Although most of the human telomeric DNA is organized into tightly spaced nucleosomes, their role in telomere protection and how they interplay with telomere-specific factors in telomere organization is still unclear. In this study we investigated whether TRF2 can regulate nucleosome assembly at telomeres.By means of chromatin immunoprecipitation (ChIP and Micrococcal Nuclease (MNase mapping assay, we found that the density of telomeric nucleosomes in human cells was inversely proportional to the dosage of TRF2 at telomeres. This effect was not observed in the G1 phase of the cell cycle but appeared coincident of late or post-replicative events. Moreover, we showed that TRF2 overexpression altered nucleosome spacing at telomeres increasing internucleosomal distance. By means of an in vitro nucleosome assembly system containing purified histones and remodeling factors, we reproduced the short nucleosome spacing found in telomeric chromatin. Importantly, when in vitro assembly was performed in the presence of purified TRF2, nucleosome spacing on a telomeric DNA template increased, in agreement with in vivo MNase mapping.Our results demonstrate that TRF2 negatively regulates the number of nucleosomes at human telomeres by a cell cycle-dependent mechanism that alters internucleosomal distance. These findings raise the intriguing possibility that telomere protection is mediated, at least in part, by the TRF2-dependent regulation of nucleosome organization.

  4. TRF2 controls telomeric nucleosome organization in a cell cycle phase-dependent manner.

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    Galati, Alessandra; Magdinier, Frédérique; Colasanti, Valentina; Bauwens, Serge; Pinte, Sébastien; Ricordy, Ruggero; Giraud-Panis, Marie-Josèphe; Pusch, Miriam Caroline; Savino, Maria; Cacchione, Stefano; Gilson, Eric

    2012-01-01

    Mammalian telomeres stabilize chromosome ends as a result of their assembly into a peculiar form of chromatin comprising a complex of non-histone proteins named shelterin. TRF2, one of the shelterin components, binds to the duplex part of telomeric DNA and is essential to fold the telomeric chromatin into a protective cap. Although most of the human telomeric DNA is organized into tightly spaced nucleosomes, their role in telomere protection and how they interplay with telomere-specific factors in telomere organization is still unclear. In this study we investigated whether TRF2 can regulate nucleosome assembly at telomeres.By means of chromatin immunoprecipitation (ChIP) and Micrococcal Nuclease (MNase) mapping assay, we found that the density of telomeric nucleosomes in human cells was inversely proportional to the dosage of TRF2 at telomeres. This effect was not observed in the G1 phase of the cell cycle but appeared coincident of late or post-replicative events. Moreover, we showed that TRF2 overexpression altered nucleosome spacing at telomeres increasing internucleosomal distance. By means of an in vitro nucleosome assembly system containing purified histones and remodeling factors, we reproduced the short nucleosome spacing found in telomeric chromatin. Importantly, when in vitro assembly was performed in the presence of purified TRF2, nucleosome spacing on a telomeric DNA template increased, in agreement with in vivo MNase mapping.Our results demonstrate that TRF2 negatively regulates the number of nucleosomes at human telomeres by a cell cycle-dependent mechanism that alters internucleosomal distance. These findings raise the intriguing possibility that telomere protection is mediated, at least in part, by the TRF2-dependent regulation of nucleosome organization.

  5. Nucleosome Positioning and Epigenetics

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    Schwab, David; Bruinsma, Robijn

    2008-03-01

    The role of chromatin structure in gene regulation has recently taken center stage in the field of epigenetics, phenomena that change the phenotype without changing the DNA sequence. Recent work has also shown that nucleosomes, a complex of DNA wrapped around a histone octamer, experience a sequence dependent energy landscape due to the variation in DNA bend stiffness with sequence composition. In this talk, we consider the role nucleosome positioning might play in the formation of heterochromatin, a compact form of DNA generically responsible for gene silencing. In particular, we discuss how different patterns of nucleosome positions, periodic or random, could either facilitate or suppress heterochromatin stability and formation.

  6. Telomere Capping Proteins are Structurally Related to RPA with an additional Telomere-Specific Domain

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    Gelinas, A.; Paschini, M; Reyes, F; Heroux, A; Batey, R; Lundblad, V; Wuttke, D

    2009-01-01

    Telomeres must be capped to preserve chromosomal stability. The conserved Stn1 and Ten1 proteins are required for proper capping of the telomere, although the mechanistic details of how they contribute to telomere maintenance are unclear. Here, we report the crystal structures of the C-terminal domain of the Saccharomyces cerevisiae Stn1 and the Schizosaccharomyces pombe Ten1 proteins. These structures reveal striking similarities to corresponding subunits in the replication protein A complex, further supporting an evolutionary link between telomere maintenance proteins and DNA repair complexes. Our structural and in vivo data of Stn1 identify a new domain that has evolved to support a telomere-specific role in chromosome maintenance. These findings endorse a model of an evolutionarily conserved mechanism of DNA maintenance that has developed as a result of increased chromosomal structural complexity.

  7. Plasmodium falciparum Nucleosomes Exhibit Reduced Stability and Lost Sequence Dependent Nucleosome Positioning.

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    Elisabeth Silberhorn

    2016-12-01

    Full Text Available The packaging and organization of genomic DNA into chromatin represents an additional regulatory layer of gene expression, with specific nucleosome positions that restrict the accessibility of regulatory DNA elements. The mechanisms that position nucleosomes in vivo are thought to depend on the biophysical properties of the histones, sequence patterns, like phased di-nucleotide repeats and the architecture of the histone octamer that folds DNA in 1.65 tight turns. Comparative studies of human and P. falciparum histones reveal that the latter have a strongly reduced ability to recognize internal sequence dependent nucleosome positioning signals. In contrast, the nucleosomes are positioned by AT-repeat sequences flanking nucleosomes in vivo and in vitro. Further, the strong sequence variations in the plasmodium histones, compared to other mammalian histones, do not present adaptations to its AT-rich genome. Human and parasite histones bind with higher affinity to GC-rich DNA and with lower affinity to AT-rich DNA. However, the plasmodium nucleosomes are overall less stable, with increased temperature induced mobility, decreased salt stability of the histones H2A and H2B and considerable reduced binding affinity to GC-rich DNA, as compared with the human nucleosomes. In addition, we show that plasmodium histone octamers form the shortest known nucleosome repeat length (155bp in vitro and in vivo. Our data suggest that the biochemical properties of the parasite histones are distinct from the typical characteristics of other eukaryotic histones and these properties reflect the increased accessibility of the P. falciparum genome.

  8. Plasmodium falciparum Nucleosomes Exhibit Reduced Stability and Lost Sequence Dependent Nucleosome Positioning

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    Silberhorn, Elisabeth; Schwartz, Uwe; Symelka, Anne; de Koning-Ward, Tania; Längst, Gernot

    2016-01-01

    The packaging and organization of genomic DNA into chromatin represents an additional regulatory layer of gene expression, with specific nucleosome positions that restrict the accessibility of regulatory DNA elements. The mechanisms that position nucleosomes in vivo are thought to depend on the biophysical properties of the histones, sequence patterns, like phased di-nucleotide repeats and the architecture of the histone octamer that folds DNA in 1.65 tight turns. Comparative studies of human and P. falciparum histones reveal that the latter have a strongly reduced ability to recognize internal sequence dependent nucleosome positioning signals. In contrast, the nucleosomes are positioned by AT-repeat sequences flanking nucleosomes in vivo and in vitro. Further, the strong sequence variations in the plasmodium histones, compared to other mammalian histones, do not present adaptations to its AT-rich genome. Human and parasite histones bind with higher affinity to GC-rich DNA and with lower affinity to AT-rich DNA. However, the plasmodium nucleosomes are overall less stable, with increased temperature induced mobility, decreased salt stability of the histones H2A and H2B and considerable reduced binding affinity to GC-rich DNA, as compared with the human nucleosomes. In addition, we show that plasmodium histone octamers form the shortest known nucleosome repeat length (155bp) in vitro and in vivo. Our data suggest that the biochemical properties of the parasite histones are distinct from the typical characteristics of other eukaryotic histones and these properties reflect the increased accessibility of the P. falciparum genome. PMID:28033404

  9. Nucleosome Packaging and Nucleosome Positioning of Genomic DNA

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    Lowary, P. T.; Widom, J.

    1997-02-01

    The goals of this study were to assess the extent to which bulk genomic DNA sequences contribute to their own packaging in nucleosomes and to reveal the relationship between nucleosome packaging and positioning. Using a competitive nucleosome reconstitution assay, we found that at least 95% of bulk DNA sequences have an affinity for histone octamer in nucleosomes that is similar to that of randomly synthesized DNA; they contribute little to their own packaging at the level of individual nucleosomes. An equation was developed that relates the measured free energy to the fractional occupancy of specific nucleosome positions. Evidently, the bulk of eukaryotic genomic DNA is also not evolved or constrained for significant sequence-directed nucleosome positioning at the level of individual nucleosomes. Implications for gene regulation in vivo are discussed.

  10. Nucleosome packaging and nucleosome positioning of genomic DNA

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    Lowary, P. T.; Widom, J

    1997-01-01

    The goals of this study were to assess the extent to which bulk genomic DNA sequences contribute to their own packaging in nucleosomes and to reveal the relationship between nucleosome packaging and positioning. Using a competitive nucleosome reconstitution assay, we found that at least 95% of bulk DNA sequences have an affinity for histone octamer in nucleosomes that is similar to that of randomly synthesized DNA; they contribute little to their own packaging at the level of individual nucle...

  11. Histone Acetylation near the Nucleosome Dyad Axis Enhances Nucleosome Disassembly by RSC and SWI/SNF.

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    Chatterjee, Nilanjana; North, Justin A; Dechassa, Mekonnen Lemma; Manohar, Mridula; Prasad, Rashmi; Luger, Karolin; Ottesen, Jennifer J; Poirier, Michael G; Bartholomew, Blaine

    2015-12-01

    Signaling associated with transcription activation occurs through posttranslational modification of histones and is best exemplified by lysine acetylation. Lysines are acetylated in histone tails and the core domain/lateral surface of histone octamers. While acetylated lysines in histone tails are frequently recognized by other factors referred to as "readers," which promote transcription, the mechanistic role of the modifications in the lateral surface of the histone octamer remains unclear. By using X-ray crystallography, we found that acetylated lysines 115 and 122 in histone H3 are solvent accessible, but in biochemical assays they appear not to interact with the bromodomains of SWI/SNF and RSC to enhance recruitment or nucleosome mobilization, as previously shown for acetylated lysines in H3 histone tails. Instead, we found that acetylation of lysines 115 and 122 increases the predisposition of nucleosomes for disassembly by SWI/SNF and RSC up to 7-fold, independent of bromodomains, and only in conjunction with contiguous nucleosomes. Thus, in combination with SWI/SNF and RSC, acetylation of lateral surface lysines in the histone octamer serves as a crucial regulator of nucleosomal dynamics distinct from the histone code readers and writers. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. Binding of NF-κB to nucleosomes: effect of translational positioning, nucleosome remodeling and linker histone H1.

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    Imtiaz Nisar Lone

    Full Text Available NF-κB is a key transcription factor regulating the expression of inflammatory responsive genes. How NF-κB binds to naked DNA templates is well documented, but how it interacts with chromatin is far from being clear. Here we used a combination of UV laser footprinting, hydroxyl footprinting and electrophoretic mobility shift assay to investigate the binding of NF-κB to nucleosomal templates. We show that NF-κB p50 homodimer is able to bind to its recognition sequence, when it is localized at the edge of the core particle, but not when the recognition sequence is at the interior of the nucleosome. Remodeling of the nucleosome by the chromatin remodeling machine RSC was not sufficient to allow binding of NF-κB to its recognition sequence located in the vicinity of the nucleosome dyad, but RSC-induced histone octamer sliding allowed clearly detectable binding of NF-κB with the slid particle. Importantly, nucleosome dilution-driven removal of H2A-H2B dimer led to complete accessibility of the site located close to the dyad to NF-κB. Finally, we found that NF-κB was able to displace histone H1 and prevent its binding to nucleosome. These data provide important insight on the role of chromatin structure in the regulation of transcription of NF-κB dependent genes.

  13. Binding of NF-κB to Nucleosomes: Effect of Translational Positioning, Nucleosome Remodeling and Linker Histone H1

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    Lone, Imtiaz Nisar; Shukla, Manu Shubhdarshan; Charles Richard, John Lalith; Peshev, Zahary Yordanov; Dimitrov, Stefan; Angelov, Dimitar

    2013-01-01

    NF-κB is a key transcription factor regulating the expression of inflammatory responsive genes. How NF-κB binds to naked DNA templates is well documented, but how it interacts with chromatin is far from being clear. Here we used a combination of UV laser footprinting, hydroxyl footprinting and electrophoretic mobility shift assay to investigate the binding of NF-κB to nucleosomal templates. We show that NF-κB p50 homodimer is able to bind to its recognition sequence, when it is localized at the edge of the core particle, but not when the recognition sequence is at the interior of the nucleosome. Remodeling of the nucleosome by the chromatin remodeling machine RSC was not sufficient to allow binding of NF-κB to its recognition sequence located in the vicinity of the nucleosome dyad, but RSC-induced histone octamer sliding allowed clearly detectable binding of NF-κB with the slid particle. Importantly, nucleosome dilution-driven removal of H2A–H2B dimer led to complete accessibility of the site located close to the dyad to NF-κB. Finally, we found that NF-κB was able to displace histone H1 and prevent its binding to nucleosome. These data provide important insight on the role of chromatin structure in the regulation of transcription of NF-κB dependent genes. PMID:24086160

  14. The size of the nucleosome

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    Bohr, Jakob; Olsen, Kasper

    2011-01-01

    The structural origin of the size of the 11 nm nucleosomal disc is addressed. On the nanometer length-scale the organization of DNA as chromatin in the chromosomes involves a coiling of DNA around the histone core of the nucleosome. We suggest that the size of the nucleosome core particle...... is dictated by the fulfillment of two criteria: One is optimizing the volume fraction of the DNA double helix; this requirement for close-packing has its root in optimizing atomic and molecular interactions. The other criterion being that of having a zero strain-twist coupling; being a zero-twist structure......-pairs of the linker-DNA is included the estimate of the size of an ideal nucleosome is in close agreement with the experimental numbers. Interestingly, the size of the nucleosome is shown to be a consequence of intrinsic properties of the DNA double helix....

  15. Consequences of cisplatin binding on nucleosome structure and dynamics.

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    Todd, Ryan C; Lippard, Stephen J

    2010-12-22

    The effects of cisplatin binding to DNA were explored at the nucleosome level to incorporate key features of the eukaryotic nuclear environment. An X-ray crystal structure of a site-specifically platinated nucleosome carrying a 1,3-cis-{Pt(NH₃)₂}²+-d(GpTpG) intrastrand cross-link reveals the details of how this adduct dictates the rotational positioning of DNA in the nucleosome. Results from in vitro nucleosome mobility assays indicate that a single platinum adduct interferes with ATP-independent sliding of DNA around the octamer core. Data from in vitro transcription experiments suggest that RNA polymerases can successfully navigate along cisplatin-damaged DNA templates that contain nucleosomes, but stall when the transcription elongation complex physically contacts a platinum cross-link located on the template strand. These results provide information about the effects of cisplatin binding to nuclear DNA and enhance our understanding of the mechanism of transcription inhibition by platinum anticancer compounds. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Nucleosome Organization in Human Embryonic Stem Cells.

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    Puya G Yazdi

    Full Text Available The fundamental repeating unit of eukaryotic chromatin is the nucleosome. Besides being involved in packaging DNA, nucleosome organization plays an important role in transcriptional regulation and cellular identity. Currently, there is much debate about the major determinants of the nucleosome architecture of a genome and its significance with little being known about its role in stem cells. To address these questions, we performed ultra-deep sequencing of nucleosomal DNA in two human embryonic stem cell lines and integrated our data with numerous epigenomic maps. Our analyses have revealed that the genome is a determinant of nucleosome organization with transcriptionally inactive regions characterized by a "ground state" of nucleosome profiles driven by underlying DNA sequences. DNA sequence preferences are associated with heterogeneous chromatin organization around transcription start sites. Transcription, histone modifications, and DNA methylation alter this "ground state" by having distinct effects on both nucleosome positioning and occupancy. As the transcriptional rate increases, nucleosomes become better positioned. Exons transcribed and included in the final spliced mRNA have distinct nucleosome profiles in comparison to exons not included at exon-exon junctions. Genes marked by the active modification H3K4m3 are characterized by lower nucleosome occupancy before the transcription start site compared to genes marked by the inactive modification H3K27m3, while bivalent domains, genes associated with both marks, lie exactly in the middle. Combinatorial patterns of epigenetic marks (chromatin states are associated with unique nucleosome profiles. Nucleosome organization varies around transcription factor binding in enhancers versus promoters. DNA methylation is associated with increasing nucleosome occupancy and different types of methylations have distinct location preferences within the nucleosome core particle. Finally, computational

  17. The structure of DNA in a nucleosome.

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    Hayes, J J; Tullius, T D; Wolffe, A P

    1990-01-01

    We describe the application of the hydroxyl radical footprinting technique to examine the histone-DNA interactions of a nucleosome that includes part of the 5S ribosomal RNA gene of Xenopus borealis. We establish that two distinct regions of DNA with different helical periodicities exist within the nucleosome and demonstrate a change in the helical periodicity of this DNA upon nucleosome formation. In particular, we find that on average the helical periodicity of DNA in this nucleosome is 10....

  18. Multiple aspects of ATP-dependent nucleosome translocation by RSC and Mi-2 are directed by the underlying DNA sequence.

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    Joke J F A van Vugt

    Full Text Available BACKGROUND: Chromosome structure, DNA metabolic processes and cell type identity can all be affected by changing the positions of nucleosomes along chromosomal DNA, a reaction that is catalysed by SNF2-type ATP-driven chromatin remodelers. Recently it was suggested that in vivo, more than 50% of the nucleosome positions can be predicted simply by DNA sequence, especially within promoter regions. This seemingly contrasts with remodeler induced nucleosome mobility. The ability of remodeling enzymes to mobilise nucleosomes over short DNA distances is well documented. However, the nucleosome translocation processivity along DNA remains elusive. Furthermore, it is unknown what determines the initial direction of movement and how new nucleosome positions are adopted. METHODOLOGY/PRINCIPAL FINDINGS: We have used AFM imaging and high resolution PAGE of mononucleosomes on 600 and 2500 bp DNA molecules to analyze ATP-dependent nucleosome repositioning by native and recombinant SNF2-type enzymes. We report that the underlying DNA sequence can control the initial direction of translocation, translocation distance, as well as the new positions adopted by nucleosomes upon enzymatic mobilization. Within a strong nucleosomal positioning sequence both recombinant Drosophila Mi-2 (CHD-type and native RSC from yeast (SWI/SNF-type repositioned the nucleosome at 10 bp intervals, which are intrinsic to the positioning sequence. Furthermore, RSC-catalyzed nucleosome translocation was noticeably more efficient when beyond the influence of this sequence. Interestingly, under limiting ATP conditions RSC preferred to position the nucleosome with 20 bp intervals within the positioning sequence, suggesting that native RSC preferentially translocates nucleosomes with 15 to 25 bp DNA steps. CONCLUSIONS/SIGNIFICANCE: Nucleosome repositioning thus appears to be influenced by both remodeler intrinsic and DNA sequence specific properties that interplay to define ATPase

  19. Relationship between nucleosome positioning and DNA methylation

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    Chodavarapu, Ramakrishna K.; Feng, Suhua; Bernatavichute, Yana V.; Chen, Pao-Yang; Stroud, Hume; Yu, Yanchun; Hetzel, Jonathan; Kuo, Frank; Kim, Jin; Cokus, Shawn J.; Casero, David; Bernal, Maria; Huijser, Peter; Clark, Amander T.; Krämer, Ute; Merchant, Sabeeha S.; Zhang, Xiaoyu; Jacobsen, Steven E.; Pellegrini, Matteo

    2010-01-01

    Nucleosomes compact and regulate access to DNA in the nucleus, and are composed of approximately 147 bases of DNA wrapped around a histone octamer1, 2. Here we report a genome-wide nucleosome positioning analysis of Arabidopsis thaliana utilizing massively parallel sequencing of mononucleosomes. By combining this data with profiles of DNA methylation at single base resolution, we identified ten base periodicities in the DNA methylation status of nucleosome-bound DNA and found that nucleosomal DNA was more highly methylated than flanking DNA. These results suggest that nucleosome positioning strongly influences DNA methylation patterning throughout the genome and that DNA methyltransferases preferentially target nucleosome-bound DNA. We also observed similar trends in human nucleosomal DNA suggesting that the relationships between nucleosomes and DNA methyltransferases are conserved. Finally, as has been observed in animals, nucleosomes were highly enriched on exons, and preferentially positioned at intron-exon and exon-intron boundaries. RNA Pol II was also enriched on exons relative to introns, consistent with the hypothesis that nucleosome positioning regulates Pol II processivity. DNA methylation is enriched on exons, consistent with the targeting of DNA methylation to nucleosomes, and suggesting a role for DNA methylation in exon definition. PMID:20512117

  20. Physics behind the mechanical nucleosome positioning code.

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    Zuiddam, Martijn; Everaers, Ralf; Schiessel, Helmut

    2017-11-01

    The positions along DNA molecules of nucleosomes, the most abundant DNA-protein complexes in cells, are influenced by the sequence-dependent DNA mechanics and geometry. This leads to the "nucleosome positioning code", a preference of nucleosomes for certain sequence motives. Here we introduce a simplified model of the nucleosome where a coarse-grained DNA molecule is frozen into an idealized superhelical shape. We calculate the exact sequence preferences of our nucleosome model and find it to reproduce qualitatively all the main features known to influence nucleosome positions. Moreover, using well-controlled approximations to this model allows us to come to a detailed understanding of the physics behind the sequence preferences of nucleosomes.

  1. A systematic analysis of nucleosome core particle and nucleosome-nucleosome stacking structure.

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    Korolev, Nikolay; Lyubartsev, Alexander P; Nordenskiöld, Lars

    2018-01-24

    Chromatin condensation is driven by the energetically favourable interaction between nucleosome core particles (NCPs). The close NCP-NCP contact, stacking, is a primary structural element of all condensed states of chromatin in vitro and in vivo. However, the molecular structure of stacked nucleosomes as well as the nature of the interactions involved in its formation have not yet been systematically studied. Here we undertake an investigation of both the structural and physico-chemical features of NCP structure and the NCP-NCP stacking. We introduce an "NCP-centred" set of parameters (NCP-NCP distance, shift, rise, tilt, and others) that allows numerical characterisation of the mutual positions of the NCPs in the stacking and in any other structures formed by the NCP. NCP stacking in more than 140 published NCP crystal structures were analysed. In addition, coarse grained (CG) MD simulations modelling NCP condensation was carried out. The CG model takes into account details of the nucleosome structure and adequately describes the long range electrostatic forces as well as excluded volume effects acting in chromatin. The CG simulations showed good agreement with experimental data and revealed the importance of the H2A and H4 N-terminal tail bridging and screening as well as tail-tail correlations in the stacked nucleosomes.

  2. Nucleosomal promoter variation generates gene expression noise.

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    Brown, Christopher R; Boeger, Hinrich

    2014-12-16

    Gene product molecule numbers fluctuate over time and between cells, confounding deterministic expectations. The molecular origins of this noise of gene expression remain unknown. Recent EM analysis of single PHO5 gene molecules of yeast indicated that promoter molecules stochastically assume alternative nucleosome configurations at steady state, including the fully nucleosomal and nucleosome-free configuration. Given that distinct configurations are unequally conducive to transcription, the nucleosomal variation of promoter molecules may constitute a source of gene expression noise. This notion, however, implies an untested conjecture, namely that the nucleosomal variation arises de novo or intrinsically (i.e., that it cannot be explained as the result of the promoter's deterministic response to variation in its molecular surroundings). Here, we show--by microscopically analyzing the nucleosome configurations of two juxtaposed physically linked PHO5 promoter copies--that the configurational variation, indeed, is intrinsically stochastic and thus, a cause of gene expression noise rather than its effect.

  3. Polymer reptation and nucleosome repositioning.

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    Schiessel, H; Widom, J; Bruinsma, R F; Gelbart, W M

    2001-05-07

    We consider how beads can diffuse along a chain that wraps them, without becoming displaced from the chain; our proposed mechanism is analogous to the reptation of "stored length" in more familiar situations of polymer dynamics. The problem arises in the case of globular aggregates of proteins (histones) that are wound by DNA in the chromosomes of plants and animals; these beads (nucleosomes) are multiply wrapped and yet are able to reposition themselves over long distances, while remaining bound by the DNA chain.

  4. Archaeal Nucleosome Positioning by CTG Repeats

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    Sandman, Kathleen; Reeve, John N.

    1999-01-01

    DNA shape recognition determines the preferred binding sites for sequence-independent DNA binding proteins, and here we document that archaeal histones assemble archaeal nucleosomes in vitro centered preferentially within (CTG)6 and (CTG)8 repeats, close to junctions with flanking mixed-sequence DNA. Archaeal nucleosomes were not positioned by (CTG)4-, (CTG)5-, or (CTG)3AA(CTG)3-containing DNA sequences. The features of CTG repeat-containing sequences that direct eucaryal nucleosome positioni...

  5. On the role of inter-nucleosomal interactions and intrinsic nucleosome dynamics in chromatin function

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    Wladyslaw A. Krajewski

    2016-03-01

    Full Text Available Evidence is emerging that many diseases result from defects in gene functions, which, in turn, depend on the local chromatin environment of a gene. However, it still remains not fully clear how chromatin activity code is ‘translated’ to the particular ‘activating’ or ‘repressing’ chromatin structural transition. Commonly, chromatin remodeling in vitro was studied using mononucleosomes as a model. However, recent data suggest that structural reorganization of a single mononucleosome is not equal to remodeling of a nucleosome particle under multinucleosomal content – such as, interaction of nucleosomes via flexible histone termini could significantly alter the mode (and the resulting products of nucleosome structural transitions. It is becoming evident that a nucleosome array does not constitute just a ‘polymer’ of individual ‘canonical’ nucleosomes due to multiple inter-nucleosomal interactions which affect nucleosome dynamics and structure. It could be hypothesized, that inter-nucleosomal interactions could act in cooperation with nucleosome inherent dynamics to orchestrate DNA-based processes and promote formation and stabilization of highly-dynamic, accessible structure of a nucleosome array. In the proposed paper we would like to discuss the nucleosome dynamics within the chromatin fiber mainly as it pertains to the roles of the structural changes mediated by inter-nucleosomal interactions.

  6. spFRET studies of nucleosome dynamics modulated by histone modifications, histone variants and neighboring nucleosomes

    NARCIS (Netherlands)

    Buning, Ruth

    2015-01-01

    At the basis of the regulation of the genetic code (DNA) in eukaryotes is its organization into nucleosomes. Nucleosomes modulate DNA accessibility through conformational dynamics like DNA breathing - the transient unwrapping of DNA from the nucleosome. Single-pair Fluorescence Resonance Energy

  7. A physical analysis of nucleosome positioning

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    Gerland, Ulrich

    2015-03-01

    The first level of genome packaging in eukaryotic cells involves the formation of dense nucleosome arrays, with DNA coverage near 90% in yeasts. A high nucleosome coverage is essential for cells, e.g. to prevent cryptic transcription, and the local positions of specific nucleosomes can play an important role in gene regulation. It is known that in vivo nucleosome positions are affected by a complex mix of passive and active mechanisms, including sequence-specific histone-DNA binding, nucleosome-nucleosome interactions, ATP-dependent remodeling enzymes, transcription, and DNA replication. Yet, the statistical distribution of nucleosome positions is extremely well described by simple physical models that treat the chromatin fiber as an interacting one-dimensional gas. I will discuss how can we interpret this surprising observation from a mechanistic perspective. I will also discuss the kinetics of the interacting gas model, which is pertinent to the question of how cells achieve the high nucleosome coverage within a short time, e.g. after DNA replication.

  8. Acetylated histone H3 increases nucleosome dissociation

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    Simon, Marek; Manohar, Mridula; Ottesen, Jennifer; Poirier, Michael

    2009-03-01

    Chromatin's basic unit structure is the nucleosome, i.e. genomic DNA wrapped around a particular class of proteins -- histones -- which due to their physical hindrance, block vital biological processes, such as DNA repair, DNA replication, and RNA transcription. Histone post-translational modifications, which are known to exist in vivo, are hypothesized to regulate these biological processes by directly altering DNA-histone interactions and thus nucleosome structure and stability. Using magnetic tweezers technique we studied the acetylation of histone H3 in the dyad region, i.e. at K115 and K122, on reconstituted arrays of nucleosomes under constant external force. Based on the measured increase in the probability of dissociation of modified nucleosomes, we infer that this double modification could facilitate histone chaperone mediated nucleosome disassembly in vivo.

  9. Characterization of Dnmt1 Binding and DNA Methylation on Nucleosomes and Nucleosomal Arrays.

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    Anna Schrader

    Full Text Available The packaging of DNA into nucleosomes and the organisation into higher order structures of chromatin limits the access of sequence specific DNA binding factors to DNA. In cells, DNA methylation is preferentially occuring in the linker region of nucleosomes, suggesting a structural impact of chromatin on DNA methylation. These observations raise the question whether DNA methyltransferases are capable to recognize the nucleosomal substrates and to modify the packaged DNA. Here, we performed a detailed analysis of nucleosome binding and nucleosomal DNA methylation by the maintenance DNA methyltransferase Dnmt1. Our binding studies show that Dnmt1 has a DNA length sensing activity, binding cooperatively to DNA, and requiring a minimal DNA length of 20 bp. Dnmt1 needs linker DNA to bind to nucleosomes and most efficiently recognizes nucleosomes with symmetric DNA linkers. Footprinting experiments reveal that Dnmt1 binds to both DNA linkers exiting the nucleosome core. The binding pattern correlates with the efficient methylation of DNA linkers. However, the enzyme lacks the ability to methylate nucleosomal CpG sites on mononucleosomes and nucleosomal arrays, unless chromatin remodeling enzymes create a dynamic chromatin state. In addition, our results show that Dnmt1 functionally interacts with specific chromatin remodeling enzymes to enable complete methylation of hemi-methylated DNA in chromatin.

  10. A positioned +1 nucleosome enhances promoter-proximal pausing

    OpenAIRE

    Jimeno-Gonz?lez, Silvia; Ceballos-Ch?vez, Mar?a; Reyes, Jos? C.

    2015-01-01

    Chromatin distribution is not uniform along the human genome. In most genes there is a promoter-associated nucleosome free region (NFR) followed by an array of nucleosomes towards the gene body in which the first (+1) nucleosome is strongly positioned. The function of this characteristic chromatin distribution in transcription is not fully understood. Here we show in vivo that the +1 nucleosome plays a role in modulating RNA polymerase II (RNAPII) promoter-proximal pausing. When a +1 nucleoso...

  11. Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro.

    Science.gov (United States)

    Hinz, John M; Laughery, Marian F; Wyrick, John J

    2015-12-08

    During Cas9 genome editing in eukaryotic cells, the bacterial Cas9 enzyme cleaves DNA targets within chromatin. To understand how chromatin affects Cas9 targeting, we characterized Cas9 activity on nucleosome substrates in vitro. We find that Cas9 endonuclease activity is strongly inhibited when its target site is located within the nucleosome core. In contrast, the nucleosome structure does not affect Cas9 activity at a target site within the adjacent linker DNA. Analysis of target sites that partially overlap with the nucleosome edge indicates that the accessibility of the protospacer-adjacent motif (PAM) is the critical determinant of Cas9 activity on a nucleosome.

  12. Human nucleosomes: special role of CG dinucleotides and Alu-nucleosomes

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    Trifonov Edward N

    2011-05-01

    Full Text Available Abstract Background The periodical occurrence of dinucleotides with a period of 10.4 bases now is undeniably a hallmark of nucleosome positioning. Whereas many eukaryotic genomes contain visible and even strong signals for periodic distribution of dinucleotides, the human genome is rather featureless in this respect. The exact sequence features in the human genome that govern the nucleosome positioning remain largely unknown. Results When analyzing the human genome sequence with the positional autocorrelation method, we found that only the dinucleotide CG shows the 10.4 base periodicity, which is indicative of the presence of nucleosomes. There is a high occurrence of CG dinucleotides that are either 31 (10.4 × 3 or 62 (10.4 × 6 base pairs apart from one another - a sequence bias known to be characteristic of Alu-sequences. In a similar analysis with repetitive sequences removed, peaks of repeating CG motifs can be seen at positions 10, 21 and 31, the nearest integers of multiples of 10.4. Conclusions Although the CG dinucleotides are dominant, other elements of the standard nucleosome positioning pattern are present in the human genome as well. The positional autocorrelation analysis of the human genome demonstrates that the CG dinucleotide is, indeed, one visible element of the human nucleosome positioning pattern, which appears both in Alu sequences and in sequences without repeats. The dominant role that CG dinucleotides play in organizing human chromatin is to indicate the involvement of human nucleosomes in tuning the regulation of gene expression and chromatin structure, which is very likely due to cytosine-methylation/-demethylation in CG dinucleotides contained in the human nucleosomes. This is further confirmed by the positions of CG-periodical nucleosomes on Alu sequences. Alu repeats appear as monomers, dimers and trimers, harboring two to six nucleosomes in a run. Considering the exceptional role CG dinucleotides play in the

  13. Tension-dependent Free Energies of Nucleosome Unwrapping

    CERN Document Server

    Lequieu, Joshua; Schwartz, David C; de Pablo, Juan J

    2016-01-01

    Nucleosomes form the basic unit of compaction within eukaryotic genomes and their locations represent an important, yet poorly understood, mechanism of genetic regulation. Quantifying the strength of interactions within the nucleosome is a central problem in biophysics and is critical to understanding how nucleosome positions influence gene expression. By comparing to single-molecule experiments, we demonstrate that a coarse-grained molecular model of the nucleosome can reproduce key aspects of nucleosome unwrapping. Using detailed simulations of DNA and histone proteins, we calculate the tension-dependent free energy surface corresponding to the unwrapping process. The model reproduces quantitatively the forces required to unwrap the nucleosome, and reveals the role played by electrostatic interactions during this process. We then demonstrate that histone modifications and DNA sequence can have significant effects on the energies of nucleosome formation. Most notably, we show that histone tails are crucial f...

  14. Dynamic nucleosome organization at hox promoters during zebrafish embryogenesis.

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    Steven E Weicksel

    Full Text Available Nucleosome organization at promoter regions plays an important role in regulating gene activity. Genome-wide studies in yeast, flies, worms, mammalian embryonic stem cells and transformed cell lines have found well-positioned nucleosomes flanking a nucleosome depleted region (NDR at transcription start sites. This nucleosome arrangement depends on DNA sequence (cis-elements as well as DNA binding factors and ATP-dependent chromatin modifiers (trans-factors. However, little is understood about how the nascent embryonic genome positions nucleosomes during development. This is particularly intriguing since the embryonic genome must undergo a broad reprogramming event upon fusion of sperm and oocyte. Using four stages of early embryonic zebrafish development, we map nucleosome positions at the promoter region of 37 zebrafish hox genes. We find that nucleosome arrangement at the hox promoters is a progressive process that takes place over several stages. At stages immediately after fertilization, nucleosomes appear to be largely disordered at hox promoter regions. At stages after activation of the embryonic genome, nucleosomes are detectable at hox promoters, with positions becoming more uniform and more highly occupied. Since the genomic sequence is invariant during embryogenesis, this progressive change in nucleosome arrangement suggests that trans-factors play an important role in organizing nucleosomes during embryogenesis. Separating hox genes into expressed and non-expressed groups shows that expressed promoters have better positioned and occupied nucleosomes, as well as distinct NDRs, than non-expressed promoters. Finally, by blocking the retinoic acid-signaling pathway, we disrupt early hox gene transcription, but observe no effect on nucleosome positions, suggesting that active hox transcription is not a driving force behind the arrangement of nucleosomes at the promoters of hox genes during early development.

  15. Stabilization of the promoter nucleosomes in nucleosome-free regions by the yeast Cyc8–Tup1 corepressor

    Science.gov (United States)

    Chen, Kaifu; Wilson, Marenda A.; Hirsch, Calley; Watson, Anjanette; Liang, Shoudan; Lu, Yue; Li, Wei; Dent, Sharon Y.R.

    2013-01-01

    The yeast Cyc8 (also known as Ssn6)–Tup1 complex regulates gene expression through a variety of mechanisms, including positioning of nucleosomes over promoters of some target genes to limit accessibility to the transcription machinery. To further define the functions of Cyc8–Tup1 in gene regulation and chromatin remodeling, we performed genome-wide profiling of changes in nucleosome organization and gene expression that occur upon loss of CYC8 or TUP1 and observed extensive nucleosome alterations in both promoters and gene bodies of derepressed genes. Our improved nucleosome profiling and analysis approaches revealed low-occupancy promoter nucleosomes (P nucleosomes) at locations previously defined as nucleosome-free regions. In the absence of CYC8 or TUP1, this P nucleosome is frequently lost, whereas nucleosomes are gained at −1 and +1 positions, accompanying up-regulation of downstream genes. Our analysis of public ChIP-seq data revealed that Cyc8 and Tup1 preferentially bind TATA-containing promoters, which are also enriched in genes derepressed upon loss of CYC8 or TUP1. These results suggest that stabilization of the P nucleosome on TATA-containing promoters may be a central feature of the repressive chromatin architecture created by the Cyc8–Tup1 corepressor, and that releasing the P nucleosome contributes to gene activation. PMID:23124522

  16. Nucleosome-coupled expression differences in closely-related species

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    Gebbia Marinella

    2011-09-01

    Full Text Available Abstract Background Genome-wide nucleosome occupancy is negatively related to the average level of transcription factor motif binding based on studies in yeast and several other model organisms. The degree to which nucleosome-motif interactions relate to phenotypic changes across species is, however, unknown. Results We address this challenge by generating nucleosome positioning and cell cycle expression data for Saccharomyces bayanus and show that differences in nucleosome occupancy reflect cell cycle expression divergence between two yeast species, S. bayanus and S. cerevisiae. Specifically, genes with nucleosome-depleted MBP1 motifs upstream of their coding sequence show periodic expression during the cell cycle, whereas genes with nucleosome-shielded motifs do not. In addition, conserved cell cycle regulatory motifs across these two species are more nucleosome-depleted compared to those that are not conserved, suggesting that the degree of conservation of regulatory sites varies, and is reflected by nucleosome occupancy patterns. Finally, many changes in cell cycle gene expression patterns across species can be correlated to changes in nucleosome occupancy on motifs (rather than to the presence or absence of motifs. Conclusions Our observations suggest that alteration of nucleosome occupancy is a previously uncharacterized feature related to the divergence of cell cycle expression between species.

  17. Exploring the Link between Nucleosome Occupancy and DNA Methylation

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    Cecilia Lövkvist

    2018-01-01

    Full Text Available Near promoters, both nucleosomes and CpG sites form characteristic spatial patterns. Previously, nucleosome depleted regions were observed upstream of transcription start sites and nucleosome occupancy was reported to correlate both with CpG density and the level of CpG methylation. Several studies imply a causal link where CpG methylation might induce nucleosome formation, whereas others argue the opposite, i.e., that nucleosome occupancy might influence CpG methylation. Correlations are indeed evident between nucleosomes, CpG density and CpG methylation—at least near promoter sites. It is however less established whether there is an immediate causal relation between nucleosome occupancy and the presence of CpG sites—or if nucleosome occupancy could be influenced by other factors. In this work, we test for such causality in human genomes by analyzing the three quantities both near and away from promoter sites. For data from the human genome we compare promoter regions with given CpG densities with genomic regions without promoters but of similar CpG densities. We find the observed correlation between nucleosome occupancy and CpG density, respectively CpG methylation, to be specific to promoter regions. In other regions along the genome nucleosome occupancy is statistically independent of the positioning of CpGs or their methylation levels. Anti-correlation between CpG density and methylation level is however similarly strong in both regions. On promoters, nucleosome occupancy is more strongly affected by the level of gene expression than CpG density or CpG methylation—calling into question any direct causal relation between nucleosome occupancy and CpG organization. Rather, our results suggest that for organisms with cytosine methylation nucleosome occupancy might be primarily linked to gene expression, with no strong impact on methylation.

  18. Nucleosome breathing and remodeling constrain CRISPR-Cas9 function.

    Science.gov (United States)

    Isaac, R Stefan; Jiang, Fuguo; Doudna, Jennifer A; Lim, Wendell A; Narlikar, Geeta J; Almeida, Ricardo

    2016-04-28

    The CRISPR-Cas9 bacterial surveillance system has become a versatile tool for genome editing and gene regulation in eukaryotic cells, yet how CRISPR-Cas9 contends with the barriers presented by eukaryotic chromatin is poorly understood. Here we investigate how the smallest unit of chromatin, a nucleosome, constrains the activity of the CRISPR-Cas9 system. We find that nucleosomes assembled on native DNA sequences are permissive to Cas9 action. However, the accessibility of nucleosomal DNA to Cas9 is variable over several orders of magnitude depending on dynamic properties of the DNA sequence and the distance of the PAM site from the nucleosome dyad. We further find that chromatin remodeling enzymes stimulate Cas9 activity on nucleosomal templates. Our findings imply that the spontaneous breathing of nucleosomal DNA together with the action of chromatin remodelers allow Cas9 to effectively act on chromatin in vivo.

  19. Enhancement of the nucleosomal pattern in sequences of lower complexity

    DEFF Research Database (Denmark)

    Bolshoy, Alexander; Shapiro, Kevin; Trifonov, Edward N.

    1997-01-01

    of a nucleosome database separated according to linguistic complexity. The pattern extracted from the subset of the simpler nucleosome sequences not only possesses all major attributes of the known nucleosomal pattern, but is substantially stronger with respect to amplitude in comparison with the total database......Intuitively, the complexity of a given DNA sequence is related to the number of various superimposed biological messages it contains. Here we assess the expectation that in nucleosome DNA sequences of lower linguistic complexity, the nucleosome DNA positioning pattern would be more pronounced than...... in those of higher linguistic complexity. The nucleosome DNA positioning pattern is one of the weakest (highly degenerate) sequence patterns. It has been extracted recently by specially designed multiple alignment procedures. We applied the most sensitive of these procedures to nearly equal subsets...

  20. Nucleosome DNA sequence structure of isochores

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    Trifonov Edward N

    2011-04-01

    Full Text Available Abstract Background Significant differences in G+C content between different isochore types suggest that the nucleosome positioning patterns in DNA of the isochores should be different as well. Results Extraction of the patterns from the isochore DNA sequences by Shannon N-gram extension reveals that while the general motif YRRRRRYYYYYR is characteristic for all isochore types, the dominant positioning patterns of the isochores vary between TAAAAATTTTTA and CGGGGGCCCCCG due to the large differences in G+C composition. This is observed in human, mouse and chicken isochores, demonstrating that the variations of the positioning patterns are largely G+C dependent rather than species-specific. The species-specificity of nucleosome positioning patterns is revealed by dinucleotide periodicity analyses in isochore sequences. While human sequences are showing CG periodicity, chicken isochores display AG (CT periodicity. Mouse isochores show very weak CG periodicity only. Conclusions Nucleosome positioning pattern as revealed by Shannon N-gram extension is strongly dependent on G+C content and different in different isochores. Species-specificity of the pattern is subtle. It is reflected in the choice of preferentially periodical dinucleotides.

  1. Genome-wide chromatin mapping with size resolution reveals a dynamic sub-nucleosomal landscape in Arabidopsis.

    Science.gov (United States)

    Pass, Daniel Antony; Sornay, Emily; Marchbank, Angela; Crawford, Margaret R; Paszkiewicz, Konrad; Kent, Nicholas A; Murray, James A H

    2017-09-01

    All eukaryotic genomes are packaged as chromatin, with DNA interlaced with both regularly patterned nucleosomes and sub-nucleosomal-sized protein structures such as mobile and labile transcription factors (TF) and initiation complexes, together forming a dynamic chromatin landscape. Whilst details of nucleosome position in Arabidopsis have been previously analysed, there is less understanding of their relationship to more dynamic sub-nucleosomal particles (subNSPs) defined as protected regions shorter than the ~150bp typical of nucleosomes. The genome-wide profile of these subNSPs has not been previously analysed in plants and this study investigates the relationship of dynamic bound particles with transcriptional control. Here we combine differential micrococcal nuclease (MNase) digestion and a modified paired-end sequencing protocol to reveal the chromatin structure landscape of Arabidopsis cells across a wide particle size range. Linking this data to RNAseq expression analysis provides detailed insight into the relationship of identified DNA-bound particles with transcriptional activity. The use of differential digestion reveals sensitive positions, including a labile -1 nucleosome positioned upstream of the transcription start site (TSS) of active genes. We investigated the response of the chromatin landscape to changes in environmental conditions using light and dark growth, given the large transcriptional changes resulting from this simple alteration. The resulting shifts in the suites of expressed and repressed genes show little correspondence to changes in nucleosome positioning, but led to significant alterations in the profile of subNSPs upstream of TSS both globally and locally. We examined previously mapped positions for the TFs PIF3, PIF4 and CCA1, which regulate light responses, and found that changes in subNSPs co-localized with these binding sites. This small particle structure is detected only under low levels of MNase digestion and is lost on more

  2. Nucleosomes Selectively Inhibit Cas9 Off-target Activity at a Site Located at the Nucleosome Edge.

    Science.gov (United States)

    Hinz, John M; Laughery, Marian F; Wyrick, John J

    2016-11-25

    Nucleosomes affect Cas9 binding and activity at on-target sites, but their impact at off-target sites is unknown. To investigate how nucleosomes affect Cas9 cleavage at off-target sites in vitro, we used a single guide RNA (sgRNA) that has been previously shown to efficiently direct Cas9 cleavage at the edge of the strongly positioned 601 nucleosome. Our data indicate that single mismatches between the sgRNA and DNA target have relatively little effect on Cas9 cleavage of naked DNA substrates, but strongly inhibit cleavage of nucleosome substrates, particularly when the mismatch is in the sgRNA "seed" region. These findings indicate that nucleosomes may enhance Cas9 specificity by inhibiting cleavage of off-target sites at the nucleosome edge. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  3. Creation of a novel telomere-cutting endonuclease based on the EN domain of telomere-specific non-long terminal repeat retrotransposon, TRAS1

    Directory of Open Access Journals (Sweden)

    Yoshitake Kazutoshi

    2010-04-01

    Full Text Available Abstract Background The ends of chromosomes, termed telomeres consist of repetitive DNA. The telomeric sequences shorten with cell division and, when telomeres are critically abbreviated, cells stop proliferating. However, in cancer cells, by the expression of telomerase which elongates telomeres, the cells can continue proliferating. Many approaches for telomere shortening have been pursued in the past, but to our knowledge, cutting telomeres in vivo has not so far been demonstrated. In addition, there is lack of information on the cellular effects of telomere shortening in human cells. Results Here, we created novel chimeric endonucleases to cut telomeres by fusing the endonuclease domain (TRAS1EN of the silkworm's telomere specific non-long terminal repeat retrotransposon TRAS1 to the human telomere-binding protein, TRF1. An in vitro assay demonstrated that the TRAS1EN-TRF1 chimeric endonucleases (T-EN and EN-T cut the human (TTAGGGn repeats specifically. The concentration of TRAS1EN-TRF1 chimeric endonucleases necessary for the cleavage of (TTAGGGn repeats was about 40-fold lower than that of TRAS1EN alone. When TRAS1EN-TRF1 endonucleases were introduced into human U2OS cancer cells using adenovirus vectors, the enzymes localized at telomeres of nuclei, cleaved and shortened the telomeric DNA by double-strand breaks. When human U2OS and HFL-1 fibroblast cells were infected with EN-T recombinant adenovirus, their cellular proliferation was suppressed for about 2 weeks after infection. In contrast, the TRAS1EN mutant (H258A chimeric endonuclease fused with TRF1 (ENmut-T did not show the suppression effect. The EN-T recombinant adenovirus induced telomere shortening in U2OS cells, activated the p53-dependent pathway and caused the senescence associated cellular responses, while the ENmut-T construct did not show such effects. Conclusions A novel TRAS1EN-TRF1 chimeric endonuclease (EN-T cuts the human telomeric repeats (TTAGGGn specifically in

  4. Nucleosomes protect DNA from DNA methylation in vivo and in vitro

    Science.gov (United States)

    Felle, Max; Hoffmeister, Helen; Rothammer, Julia; Fuchs, Andreas; Exler, Josef H.; Längst, Gernot

    2011-01-01

    Positioned nucleosomes limit the access of proteins to DNA. However, the impact of nucleosomes on DNA methylation in vitro and in vivo is poorly understood. Here, we performed a detailed analysis of nucleosome binding and nucleosomal DNA methylation by the de novo methyltransferases. We show that compared to linker DNA, nucleosomal DNA is largely devoid of CpG methylation. ATP-dependent chromatin remodelling frees nucleosomal CpG dinucleotides and renders the remodelled nucleosome a 2-fold better substrate for Dnmt3a methyltransferase compared to free DNA. These results reflect the situation in vivo, as quantification of nucleosomal DNA methylation levels in HeLa cells shows a 2-fold decrease of nucleosomal DNA methylation levels compared to linker DNA. Our findings suggest that nucleosomal positions are stably maintained in vivo and nucleosomal occupancy is a major determinant of global DNA methylation patterns in vivo. PMID:21622955

  5. Theoretical analysis of epigenetic cell memory by nucleosome modification

    DEFF Research Database (Denmark)

    Dodd, Ian B; Micheelsen, Mille A; Sneppen, Kim

    2007-01-01

    strong bistability that is resistant both to high noise due to random gain or loss of nucleosome modifications and to random partitioning upon DNA replication. However, robust bistability required: (1) cooperativity, the activity of more than one modified nucleosome, in the modification reactions and (2...

  6. Analysis of Nucleosome Transcription Using Single-Particle FRET

    Science.gov (United States)

    Feofanov, Alexey V.; Kudryashova, Kseniya S.; Chertkov, Oleg V.; Nikitin, Dmitry V.; Pestov, Nikolai A.; Kulaeva, Olga I.; Studitsky, Vasily M.; Kirpichnikov, Mikhail P.

    Many biological reactions including transcription of a gene are too complex and heterogeneous to be understood by studying ensembles of interacting molecules. In these cases analysis of single complexes can clarify structural and dynamic aspects of these processes. Here we report that single-particle Förster resonance energy transfer (spFRET ) microscopy is applicable to investigation of transcription through nucleosomes by an RNA polymerase . Mononucleosomes that support transcription were assembled from core histones and short DNA containing the T7A1 promoter and strong 603 nucleosome-positioning sequence. Fluorophores (Cy3 and Cy5) were introduced in the neighboring coils of nucleosome DNA in spatially close positions without disturbance of nucleosomal structure or transcription. Such labeling allows the changes in the Cy3-Cy5 distance caused by DNA uncoiling from the octamer or DNA looping to be monitored as changes in FRET efficiency. spFRET measurements for freely diffusing single nucleosomes were conducted using a laser scanning confocal microscope equipped with avalanche photodiodes. Nucleosome subpopulations that differ in FRET efficiency (i.e. in nucleosome structure) were revealed. RNA polymerase was stalled in distinct positions on the nucleosomal DNA during transcription, and the structures of these complexes were characterized with spFRET microscopy.

  7. Hydroxymethylation of DNA influences nucleosomal conformation and stability in vitro.

    Science.gov (United States)

    Mendonca, Agnes; Chang, En Hyung; Liu, Wenjie; Yuan, Chongli

    2014-11-01

    Hydroxymethylation of DNA at the C5 position of cytosine (5hmC) is recognized as an important epigenetic mark. The molecular role of 5hmC in gene regulation, however, is not well understood. We studied the effects of 5-hydroxymethylation (5hmC) on nucleosome properties in vitro using a combination of biochemical and fluorescence assays. Competitive reconstitution was used to evaluate the effect of 5hmC on nucleosome formation. The effects of 5hmC on nucleosome compactness and stability were characterized using FRET assays. These findings have also been compared with another important epigenetic mark, the cytosine methylation (5mC) of DNA. We observed that hydroxymethylation increases the binding affinity of DNA for the histone octamer. The formed nucleosome exhibits slightly different conformations based on the sequence and epigenetic context of DNA. DNA hydroxymethylation decreases the stability of formed nucleosomes in salt-induced dissociation processes. DNA containing 5hmC is more likely to be incorporated into nucleosomes. Once formed, the 5hmC nucleosomes might be in an open and transcriptionally active state due to the weakened interaction of hydroxymethylated DNA with the H2A-H2B dimers. Our results reveal the effect of 5hmC on regulating nucleosome compactness and stability in vitro. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Uncovering the forces between nucleosomes using DNA origami.

    Science.gov (United States)

    Funke, Jonas J; Ketterer, Philip; Lieleg, Corinna; Schunter, Sarah; Korber, Philipp; Dietz, Hendrik

    2016-11-01

    Revealing the energy landscape for nucleosome association may contribute to the understanding of higher-order chromatin structures and their impact on genome regulation. We accomplish this in a direct measurement by integrating two nucleosomes into a DNA origami-based force spectrometer, which enabled subnanometer-resolution measurements of nucleosome-nucleosome distance frequencies via single-particle electron microscopy imaging. From the data, we derived the Boltzmann-weighted distance-dependent energy landscape for nucleosome pair interactions. We find a shallow but long-range (~6 nm) attractive nucleosome pair potential with a minimum of -1.6 kcal/mol close to direct contact distances. The relative nucleosome orientation had little influence, but histone H4 acetylation or removal of histone tails drastically decreased the interaction strength. Because of the weak and shallow pair potential, higher-order nucleosome assemblies will be compliant and experience dynamic shape fluctuations in the absence of additional cofactors. Our results contribute to a more accurate description of chromatin and our force spectrometer provides a powerful tool for the direct and high-resolution study of molecular interactions using imaging techniques.

  9. Sequence signatures of nucleosome positioning in Caenorhabditis elegans.

    Science.gov (United States)

    Chen, Kaifu; Wang, Lei; Yang, Meng; Liu, Jiucheng; Xin, Chengqi; Hu, Songnian; Yu, Jun

    2010-06-01

    Our recent investigation in the protist Trichomonas vaginalis suggested a DNA sequence periodicity with a unit length of 120.9 nt, which represents a sequence signature for nucleosome positioning. We now extended our observation in higher eukaryotes and identified a similar periodicity of 175 nt in length in Caenorhabditis elegans. In the process of defining the sequence compositional characteristics, we found that the 10.5-nt periodicity, the sequence signature of DNA double helix, may not be sufficient for cross-nucleosome positioning but provides essential guiding rails to facilitate positioning. We further dissected nucleosome-protected sequences and identified a strong positive purine (AG) gradient from the 5'-end to the 3'-end, and also learnt that the nucleosome-enriched regions are GC-rich as compared to the nucleosome-free sequences as purine content is positively correlated with GC content. Sequence characterization allowed us to develop a hidden Markov model (HMM) algorithm for decoding nucleosome positioning computationally, and based on a set of training data from the fifth chromosome of C. elegans, our algorithm predicted 60%-70% of the well-positioned nucleosomes, which is 15%-20% higher than random positioning. We concluded that nucleosomes are not randomly positioned on DNA sequences and yet bind to different genome regions with variable stability, well-positioned nucleosomes leave sequence signatures on DNA, and statistical positioning of nucleosomes across genome can be decoded computationally based on these sequence signatures. Copyright 2010 Beijing Genomics Institute. Published by Elsevier Ltd. All rights reserved.

  10. Dynamics of Nucleosome Positioning Maturation following Genomic Replication

    Directory of Open Access Journals (Sweden)

    Pauline Vasseur

    2016-09-01

    Full Text Available Chromatin is thought to carry epigenetic information from one generation to the next, although it is unclear how such information survives the disruptions of nucleosomal architecture occurring during genomic replication. Here, we measure a key aspect of chromatin structure dynamics during replication—how rapidly nucleosome positions are established on the newly replicated daughter genomes. By isolating newly synthesized DNA marked with 5-ethynyl-2′-deoxyuridine (EdU, we characterize nucleosome positions on both daughter genomes of S. cerevisiae during chromatin maturation. We find that nucleosomes rapidly adopt their mid-log positions at highly transcribed genes, which is consistent with a role for transcription in positioning nucleosomes in vivo. Additionally, experiments in hir1Δ mutants reveal a role for HIR in nucleosome spacing. We also characterized nucleosome positions on the leading and lagging strands, uncovering differences in chromatin maturation dynamics at hundreds of genes. Our data define the maturation dynamics of newly replicated chromatin and support a role for transcription in sculpting the chromatin template.

  11. ISWI chromatin remodellers sense nucleosome modifications to determine substrate preference

    DEFF Research Database (Denmark)

    Dann, Geoffrey P; Liszczak, Glen P; Bagert, John D

    2017-01-01

    ATP-dependent chromatin remodellers regulate access to genetic information by controlling nucleosome positions in vivo. However, the mechanism by which remodellers discriminate between different nucleosome substrates is poorly understood. Many chromatin remodelling proteins possess conserved...... activity of all ISWI remodellers evaluated. This dependence also extends to CHD and SWI/SNF family remodellers, suggesting that the acidic patch may be generally required for chromatin remodelling. Critically, remodelling activity can be regulated by modifications neighbouring the acidic patch, signifying...... that it may act as a tunable interaction hotspot for ATP-dependent chromatin remodellers and, by extension, many other chromatin effectors that engage this region of the nucleosome surface....

  12. Repertoires of the nucleosome-positioning dinucleotides.

    Directory of Open Access Journals (Sweden)

    Thomas Bettecken

    Full Text Available It is generally accepted that the organization of eukaryotic DNA into chromatin is strongly governed by a code inherent in the genomic DNA sequence. This code, as well as other codes, is superposed on the triplets coding for amino acids. The history of the chromatin code started three decades ago with the discovery of the periodic appearance of certain dinucleotides, with AA/TT and RR/YY giving the strongest signals, all with a period of 10.4 bases. Every base-pair stack in the DNA duplex has specific deformation properties, thus favoring DNA bending in a specific direction. The appearance of the corresponding dinucleotide at the distance 10.4 xn bases will facilitate DNA bending in that direction, which corresponds to the minimum energy of DNA folding in the nucleosome. We have analyzed the periodic appearances of all 16 dinucleotides in the genomes of thirteen different eukaryotic organisms. Our data show that a large variety of dinucleotides (if not all are, apparently, contributing to the nucleosome positioning code. The choice of the periodical dinucleotides differs considerably from one organism to another. Among other 10.4 base periodicities, a strong and very regular 10.4 base signal was observed for CG dinucleotides in the genome of the honey bee A. mellifera. Also, the dinucleotide CG appears as the only periodical component in the human genome. This observation seems especially relevant since CpG methylation is well known to modulate chromatin packing and regularity. Thus, the selection of the dinucleotides contributing to the chromatin code is species specific, and may differ from region to region, depending on the sequence context.

  13. DNase I footprinting of the nucleosome in whole nuclei.

    Science.gov (United States)

    Staynov, D Z

    2008-07-18

    DNase I was used to footprint the 147 bp DNA fragment of the nucleosome in whole chicken erythrocyte nuclei. It was found that the higher-order structure imposes an additional protection on nucleosomes at sites close to the entry and exit points of the linker DNA, around the dyad axis (site S 0). The observed protection is extended up to 20 bp on either side of S 0. It is partial (approximately 50%) and most probably reflects a full protection of different regions in alternatively oriented nucleosomes. These are the same regions which interact with linker histones. The results strongly support the findings by simulation of DNase I digests of unlabelled oligonucleosome fragments in the 30 nm fibre that in all nucleosomes sites S -5 to S -3 and S +3 to S +5 ara on the outside of the fibre exposed to DNase I.

  14. Nucleosome Positioning and NDR Structure at RNA Polymerase III Promoters

    DEFF Research Database (Denmark)

    Helbo, Alexandra Søgaard; Lay, Fides D; Jones, Peter A

    2017-01-01

    Chromatin is structurally involved in the transcriptional regulation of all genes. While the nucleosome positioning at RNA polymerase II (pol II) promoters has been extensively studied, less is known about the chromatin structure at pol III promoters in human cells. We use a high......-resolution analysis to show substantial differences in chromatin structure of pol II and pol III promoters, and between subtypes of pol III genes. Notably, the nucleosome depleted region at the transcription start site of pol III genes extends past the termination sequences, resulting in nucleosome free gene bodies...... the first high-resolution map of nucleosome positioning and occupancy at human pol III promoters at specific loci and genome wide....

  15. Hidden Markov Analysis of Nucleosome Unwrapping Under Force

    National Research Council Canada - National Science Library

    Kruithof, M; van Noort, J

    2009-01-01

    .... Here we measured the force-induced unwrapping of DNA from a single nucleosome and show that hidden Markov analysis, adopted for the nonlinear force-extension of DNA, can readily resolve unwrapping...

  16. The universality of nucleosome organization: from yeast to human

    Science.gov (United States)

    Chereji, Razvan

    The basic units of DNA packaging are called nucleosomes. Their locations on the chromosomes play an essential role in gene regulation. We study nucleosome positioning in yeast, fly, mouse, and human, and build biophysical models in order to explain the genome-wide nucleosome organization. We show that DNA sequence alone is not able to generate the phased arrays of nucleosomes observed in vivo near the transcription start sites. We discuss simple models which can account for the formation of nucleosome depleted regions and nucleosome phasing at the gene promoters. We show that the same principles apply to different organisms. References: [1] RV Chereji, D Tolkunov, G Locke, AV Morozov - Phys. Rev. E 83, 050903 (2011) [2] RV Chereji, AV Morozov - J. Stat. Phys. 144, 379 (2011) [3] RV Chereji, AV Morozov - Proc. Natl. Acad. Sci. U.S.A. 111, 5236 (2014) [4] RV Chereji, T-W Kan, et al. - Nucleic Acids Res. (2015) doi: 10.1093/nar/gkv978 [5] RV Chereji, AV Morozov - Brief. Funct. Genomics 14, 50 (2015) [6] HA Cole, J Ocampo, JR Iben, RV Chereji, DJ Clark - Nucleic Acids Res. 42, 12512 (2014) [7] D Ganguli, RV Chereji, J Iben, HA Cole, DJ Clark - Genome Res. 24, 1637 (2014)

  17. Single-nucleosome mapping of histone modifications in S. cerevisiae.

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    Chih Long Liu

    2005-10-01

    Full Text Available Covalent modification of histone proteins plays a role in virtually every process on eukaryotic DNA, from transcription to DNA repair. Many different residues can be covalently modified, and it has been suggested that these modifications occur in a great number of independent, meaningful combinations. Published low-resolution microarray studies on the combinatorial complexity of histone modification patterns suffer from confounding effects caused by the averaging of modification levels over multiple nucleosomes. To overcome this problem, we used a high-resolution tiled microarray with single-nucleosome resolution to investigate the occurrence of combinations of 12 histone modifications on thousands of nucleosomes in actively growing S. cerevisiae. We found that histone modifications do not occur independently; there are roughly two groups of co-occurring modifications. One group of lysine acetylations shows a sharply defined domain of two hypo-acetylated nucleosomes, adjacent to the transcriptional start site, whose occurrence does not correlate with transcription levels. The other group consists of modifications occurring in gradients through the coding regions of genes in a pattern associated with transcription. We found no evidence for a deterministic code of many discrete states, but instead we saw blended, continuous patterns that distinguish nucleosomes at one location (e.g., promoter nucleosomes from those at another location (e.g., over the 3' ends of coding regions. These results are consistent with the idea of a simple, redundant histone code, in which multiple modifications share the same role.

  18. Role of nucleosome remodeling in neurodevelopmental and intellectual disability disorders

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    Alberto J Lopez

    2015-04-01

    Full Text Available It is becoming increasingly important to understand how epigenetic mechanisms control gene expression during neurodevelopment. Two epigenetic mechanisms that have received considerable attention are DNA methylation and histone acetylation. Human exome sequencing and genome-wide association studies have linked several neurobiological disorders to genes whose products actively regulate DNA methylation and histone acetylation. More recently, a third major epigenetic mechanism, nucleosome remodeling, has been implicated in human developmental and intellectual disability disorders. Nucleosome remodeling is driven primarily through nucleosome remodeling complexes with specialized ATP-dependent enzymes. These enzymes directly interact with DNA or chromatin structure, as well as histone subunits, to restructure the shape and organization of nucleosome positioning to ultimately regulate gene expression. Of particular interest is the neuron-specific Brg1/hBrm Associated Factor (nBAF complex. Mutations in nBAF subunit genes have so far been linked to Coffin-Siris syndrome, Nicolaides-Baraitser syndrome, schizophrenia, and Autism Spectrum Disorder. Together, these human developmental and intellectual disability disorders are powerful examples of the impact of epigenetic modulation on gene expression. This review focuses on the new and emerging role of nucleosome remodeling in neurodevelopmental and intellectual disability disorders and whether nucleosome remodeling affects gene expression required for cognition independently of its role in regulating gene expression required for development.

  19. Multiple distinct stimuli increase measured nucleosome occupancy around human promoters.

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    Chuong D Pham

    Full Text Available Nucleosomes can block access to transcription factors. Thus the precise localization of nucleosomes relative to transcription start sites and other factor binding sites is expected to be a critical component of transcriptional regulation. Recently developed microarray approaches have allowed the rapid mapping of nucleosome positions over hundreds of kilobases (kb of human genomic DNA, although these approaches have not yet been widely used to measure chromatin changes associated with changes in transcription. Here, we use custom tiling microarrays to reveal changes in nucleosome positions and abundance that occur when hormone-bound glucocorticoid receptor (GR binds to sites near target gene promoters in human osteosarcoma cells. The most striking change is an increase in measured nucleosome occupancy at sites spanning ∼1 kb upstream and downstream of transcription start sites, which occurs one hour after addition of hormone, but is lost at 4 hours. Unexpectedly, this increase was seen both on GR-regulated and GR-non-regulated genes. In addition, the human SWI/SNF chromatin remodeling factor (a GR co-activator was found to be important for increased occupancy upon hormone treatment and also for low nucleosome occupancy without hormone. Most surprisingly, similar increases in nucleosome occupancy were also seen on both regulated and non-regulated promoters during differentiation of human myeloid leukemia cells and upon activation of human CD4+ T-cells. These results indicate that dramatic changes in chromatin structure over ∼2 kb of human promoters may occur genomewide and in response to a variety of stimuli, and suggest novel models for transcriptional regulation.

  20. Phase diagram of nucleosome core particles.

    Science.gov (United States)

    Mangenot, S; Leforestier, A; Durand, D; Livolant, F

    2003-11-07

    We present a phase diagram of the nucleosome core particle (NCP) as a function of the monovalent salt concentration and applied osmotic pressure. Above a critical pressure, NCPs stack on top of each other to form columns that further organize into multiple columnar phases. An isotropic (and in some cases a nematic) phase of columns is observed in the moderate pressure range. Under higher pressure conditions, a lamello-columnar phase and an inverse hexagonal phase form under low salt conditions, whereas a 2D hexagonal phase or a 3D orthorhombic phase is found at higher salt concentration. For intermediate salt concentrations, microphase separation occurs. The richness of the phase diagram originates from the heterogeneous distribution of charges at the surface of the NCP, which makes the particles extremely sensitive to small ionic variations of their environment, with consequences on their interactions and supramolecular organization. We discuss how the polymorphism of NCP supramolecular organization may be involved in chromatin changes in the cellular context.

  1. Mobilities

    DEFF Research Database (Denmark)

    to social networks, personal identities, and our relationship to the built environment. The omnipresence of mobilities within everyday life, high politics, technology, and tourism (to mention but a few) all point to a key insight harnessed by the ‘mobilities turn’. Namely that mobilities is much more than...... and environmental degradation. The spaces and territories marked by mobilities as well as the sites marked by the bypassing of such are explored. Moreover, the architectural and technological dimensions to infrastructures and sites of mobilities will be included as well as the issues of power, social exclusion......, consumption, surveillance and mobilities history to mention some of the many themes covered by this reference work. This new title will focus on the academic contributions to this understanding by primarily focusing on works and publications in the aftermath of the seminal book and landmark text ‘Sociology...

  2. Structure of the CENP-A nucleosome and its implications for centromeric chromatin architecture.

    Science.gov (United States)

    Tachiwana, Hiroaki; Kurumizaka, Hitoshi

    2011-01-01

    Centromeres are dictated by the epigenetic inheritance of the centromeric nucleosome containing the centromere-specific histone H3 variant, CENP-A. The structure of the CENP-A nucleosome has been considered to be the fundamental architecture of the centromeric chromatin. Controversy exists in the literature regarding the CENP-A nucleosome structures, with octasome, hemisome, compact octasome, hexasome, and tetrasome models being reported. Some of these CENP-A nucleosome models may correspond to transient intermediates for the assembly of the mature CENP-A nucleosome; however, their significances are still unclear. Therefore, the structure of the mature CENP-A nucleosome has been eagerly awaited. We reconstituted the human CENP-A nucleosome with its cognate centromeric DNA fragment, and determined its crystal structure. In this review, we describe the structure and the physical properties of the CENP-A nucleosome, and discuss their implications for centromeric chromatin architecture.

  3. Structural Mechanisms of Nucleosome Recognition by Linker Histones.

    Science.gov (United States)

    Zhou, Bing-Rui; Jiang, Jiansheng; Feng, Hanqiao; Ghirlando, Rodolfo; Xiao, T Sam; Bai, Yawen

    2015-08-20

    Linker histones bind to the nucleosome and regulate the structure of chromatin and gene expression. Despite more than three decades of effort, the structural basis of nucleosome recognition by linker histones remains elusive. Here, we report the crystal structure of the globular domain of chicken linker histone H5 in complex with the nucleosome at 3.5 Å resolution, which is validated using nuclear magnetic resonance spectroscopy. The globular domain sits on the dyad of the nucleosome and interacts with both DNA linkers. Our structure integrates results from mutation analyses and previous cross-linking and fluorescence recovery after photobleach experiments, and it helps resolve the long debate on structural mechanisms of nucleosome recognition by linker histones. The on-dyad binding mode of the H5 globular domain is different from the recently reported off-dyad binding mode of Drosophila linker histone H1. We demonstrate that linker histones with different binding modes could fold chromatin to form distinct higher-order structures. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. The nucleosome: orchestrating DNA damage signaling and repair within chromatin.

    Science.gov (United States)

    Agarwal, Poonam; Miller, Kyle M

    2016-10-01

    DNA damage occurs within the chromatin environment, which ultimately participates in regulating DNA damage response (DDR) pathways and repair of the lesion. DNA damage activates a cascade of signaling events that extensively modulates chromatin structure and organization to coordinate DDR factor recruitment to the break and repair, whilst also promoting the maintenance of normal chromatin functions within the damaged region. For example, DDR pathways must avoid conflicts between other DNA-based processes that function within the context of chromatin, including transcription and replication. The molecular mechanisms governing the recognition, target specificity, and recruitment of DDR factors and enzymes to the fundamental repeating unit of chromatin, i.e., the nucleosome, are poorly understood. Here we present our current view of how chromatin recognition by DDR factors is achieved at the level of the nucleosome. Emerging evidence suggests that the nucleosome surface, including the nucleosome acidic patch, promotes the binding and activity of several DNA damage factors on chromatin. Thus, in addition to interactions with damaged DNA and histone modifications, nucleosome recognition by DDR factors plays a key role in orchestrating the requisite chromatin response to maintain both genome and epigenome integrity.

  5. Chemical map of Schizosaccharomyces pombe reveals species-specific features in nucleosome positioning

    Science.gov (United States)

    Moyle-Heyrman, Georgette; Zaichuk, Tetiana; Xi, Liqun; Zhang, Quanwei; Uhlenbeck, Olke C.; Holmgren, Robert; Widom, Jonathan; Wang, Ji-Ping

    2013-01-01

    Using a recently developed chemical approach, we have generated a genome-wide map of nucleosomes in vivo in Schizosaccharomyces pombe (S. pombe) at base pair resolution. The shorter linker length previously identified in S. pombe is due to a preponderance of nucleosomes separated by ∼4/5 bp, placing nucleosomes on opposite faces of the DNA. The periodic dinucleotide feature thought to position nucleosomes is equally strong in exons as in introns, demonstrating that nucleosome positioning information can be superimposed on coding information. Unlike the case in Saccharomyces cerevisiae, A/T-rich sequences are enriched in S. pombe nucleosomes, particularly at ±20 bp around the dyad. This difference in nucleosome binding preference gives rise to a major distinction downstream of the transcription start site, where nucleosome phasing is highly predictable by A/T frequency in S. pombe but not in S. cerevisiae, suggesting that the genomes and DNA binding preferences of nucleosomes have coevolved in different species. The poly (dA-dT) tracts affect but do not deplete nucleosomes in S. pombe, and they prefer special rotational positions within the nucleosome, with longer tracts enriched in the 10- to 30-bp region from the dyad. S. pombe does not have a well-defined nucleosome-depleted region immediately upstream of most transcription start sites; instead, the −1 nucleosome is positioned with the expected spacing relative to the +1 nucleosome, and its occupancy is negatively correlated with gene expression. Although there is generally very good agreement between nucleosome maps generated by chemical cleavage and micrococcal nuclease digestion, the chemical map shows consistently higher nucleosome occupancy on DNA with high A/T content. PMID:24277842

  6. Nature of the Nucleosomal Barrier to RNA Polymerase II | Center for Cancer Research

    Science.gov (United States)

    In the cell, RNA polymerase II (pol II) efficiently transcribes DNA packaged into nucleosomes, but in vitro encounters with the nucleosomes induce catalytic inactivation (arrest) of the pol II core enzyme. To determine potential mechanisms making nucleosomes transparent to transcription in vivo, we analyzed the nature of the nucleosome-induced arrest. We found that the arrests have been detected mostly at positions of strong intrinsic pause sites of DNA.

  7. Reactivity in ELISA with DNA-loaded nucleosomes in patients with proliferative lupus nephritis

    NARCIS (Netherlands)

    Dieker, J.W.; Schlumberger, W.; McHugh, N.; Hamann, P.; Vlag, J. van der; Berden, J.H.M.

    2015-01-01

    Autoantibodies against nucleosomes are considered a hallmark of systemic lupus erythematosus (SLE). We compared in patients with proliferative lupus nephritis the diagnostic usefulness of a dsDNA-loaded nucleosome ELISA (anti-dsDNA-NcX) with ELISAs in which dsDNA or nucleosomes alone were coated.

  8. The Chd1 Chromatin Remodeler Shifts Nucleosomal DNA Bidirectionally as a Monomer

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    Qiu, Yupeng; Levendosky, Robert F.; Chakravarthy, Srinivas; Patel, Ashok; Bowman, Gregory D.; Myong, Sua

    2017-10-01

    Chromatin remodelers catalyze dynamic packaging of the genome by carrying out nucleosome assembly/disassembly, histone exchange, and nucleosome repositioning. Remodeling results in evenly spaced nucleosomes, which requires probing both sides of the nucleosome, yet the way remodelers organize sliding activity to achieve this task is not understood. Here, we show that the monomeric Chd1 remodeler shifts DNA back and forth by dynamically alternating between different segments of the nucleosome. During sliding, Chd1 generates unstable remodeling intermediates that spontaneously relax to a pre-remodeled position. We demonstrate that nucleosome sliding is tightly controlled by two regulatory domains: the DNA-binding domain, which interferes with sliding when its range is limited by a truncated linking segment, and the chromodomains, which play a key role in substrate discrimination. We propose that active interplay of the ATPase motor with the regulatory domains may promote dynamic nucleosome structures uniquely suited for histone exchange and chromatin reorganization during transcription.

  9. Dynamics of nucleosome assembly and effects of DNA methylation.

    Science.gov (United States)

    Lee, Ju Yeon; Lee, Jaehyoun; Yue, Hongjun; Lee, Tae-Hee

    2015-02-13

    The nucleosome is the fundamental packing unit of the eukaryotic genome, and CpG methylation is an epigenetic modification associated with gene repression and silencing. We investigated nucleosome assembly mediated by histone chaperone Nap1 and the effects of CpG methylation based on three-color single molecule FRET measurements, which enabled direct monitoring of histone binding in the context of DNA wrapping. According to our observation, (H3-H4)2 tetramer incorporation must precede H2A-H2B dimer binding, which is independent of DNA termini wrapping. Upon CpG methylation, (H3-H4)2 tetramer incorporation and DNA termini wrapping are facilitated, whereas proper incorporation of H2A-H2B dimers is inhibited. We suggest that these changes are due to rigidified DNA and increased random binding of histones to DNA. According to the results, CpG methylation expedites nucleosome assembly in the presence of abundant DNA and histones, which may help facilitate gene packaging in chromatin. The results also indicate that the slowest steps in nucleosome assembly are DNA termini wrapping and tetramer positioning, both of which are affected heavily by changes in the physical properties of DNA. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  10. Propagation of thrombosis by neutrophils and extracellular nucleosome networks.

    Science.gov (United States)

    Pfeiler, Susanne; Stark, Konstantin; Massberg, Steffen; Engelmann, Bernd

    2017-02-01

    Neutrophils, early mediators of the innate immune defense, are recruited to developing thrombi in different types of thrombosis. They amplify intravascular coagulation by stimulating the tissue factor-dependent extrinsic pathway via inactivation of endogenous anticoagulants, enhancing factor XII activation or decreasing plasmin generation. Neutrophil-dependent prothrombotic mechanisms are supported by the externalization of decondensed nucleosomes and granule proteins that together form neutrophil extracellular traps. These traps, either in intact or fragmented form, are causally involved in various forms of experimental thrombosis as first indicated by their role in the enhancement of both microvascular thrombosis during bacterial infection and carotid artery thrombosis. Neutrophil extracellular traps can be induced by interactions of neutrophils with activated platelets; vice versa, these traps enhance adhesion of platelets via von Willebrand factor. Neutrophil-induced microvascular thrombus formation can restrict the dissemination and survival of blood-borne bacteria and thereby sustain intravascular immunity. Dysregulation of this innate immune pathway may support sepsis-associated coagulopathies. Notably, neutrophils and extracellular nucleosomes, together with platelets, critically promote fibrin formation during flow restriction-induced deep vein thrombosis. Neutrophil extracellular traps/extracellular nucleosomes are increased in thrombi and in the blood of patients with different vaso-occlusive pathologies and could be therapeutically targeted for the prevention of thrombosis. Thus, during infections and in response to blood vessel damage, neutrophils and externalized nucleosomes are major promoters of intravascular blood coagulation and thrombosis. Copyright© Ferrata Storti Foundation.

  11. Transcription and Remodeling Produce Asymmetrically Unwrapped Nucleosomal Intermediates.

    Science.gov (United States)

    Ramachandran, Srinivas; Ahmad, Kami; Henikoff, Steven

    2017-12-21

    Nucleosomes are disrupted during transcription and other active processes, but the structural intermediates during nucleosome disruption in vivo are unknown. To identify intermediates, we mapped subnucleosomal protections in Drosophila cells using Micrococcal Nuclease followed by sequencing. At the first nucleosome position downstream of the transcription start site, we identified unwrapped intermediates, including hexasomes that lack either proximal or distal contacts. Inhibiting topoisomerases or depleting histone chaperones increased unwrapping, whereas inhibiting release of paused RNAPII or reducing RNAPII elongation decreased unwrapping. Our results indicate that positive torsion generated by elongating RNAPII causes transient loss of histone-DNA contacts. Using this mapping approach, we found that nucleosomes flanking human CTCF insulation sites are similarly disrupted. We also identified diagnostic subnucleosomal particle remnants in cell-free human DNA data as a relic of transcribed genes from apoptosing cells. Thus identification of subnucleosomal fragments from nuclease protection data represents a general strategy for structural epigenomics. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Comparative analysis of methods for genome-wide nucleosome cartography.

    Science.gov (United States)

    Quintales, Luis; Vázquez, Enrique; Antequera, Francisco

    2015-07-01

    Nucleosomes contribute to compacting the genome into the nucleus and regulate the physical access of regulatory proteins to DNA either directly or through the epigenetic modifications of the histone tails. Precise mapping of nucleosome positioning across the genome is, therefore, essential to understanding the genome regulation. In recent years, several experimental protocols have been developed for this purpose that include the enzymatic digestion, chemical cleavage or immunoprecipitation of chromatin followed by next-generation sequencing of the resulting DNA fragments. Here, we compare the performance and resolution of these methods from the initial biochemical steps through the alignment of the millions of short-sequence reads to a reference genome to the final computational analysis to generate genome-wide maps of nucleosome occupancy. Because of the lack of a unified protocol to process data sets obtained through the different approaches, we have developed a new computational tool (NUCwave), which facilitates their analysis, comparison and assessment and will enable researchers to choose the most suitable method for any particular purpose. NUCwave is freely available at http://nucleosome.usal.es/nucwave along with a step-by-step protocol for its use. © The Author 2014. Published by Oxford University Press. For Permissions, please email: journals.permissions@oup.com.

  13. Structural features based genome-wide characterization and prediction of nucleosome organization

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    Gan Yanglan

    2012-03-01

    Full Text Available Abstract Background Nucleosome distribution along chromatin dictates genomic DNA accessibility and thus profoundly influences gene expression. However, the underlying mechanism of nucleosome formation remains elusive. Here, taking a structural perspective, we systematically explored nucleosome formation potential of genomic sequences and the effect on chromatin organization and gene expression in S. cerevisiae. Results We analyzed twelve structural features related to flexibility, curvature and energy of DNA sequences. The results showed that some structural features such as DNA denaturation, DNA-bending stiffness, Stacking energy, Z-DNA, Propeller twist and free energy, were highly correlated with in vitro and in vivo nucleosome occupancy. Specifically, they can be classified into two classes, one positively and the other negatively correlated with nucleosome occupancy. These two kinds of structural features facilitated nucleosome binding in centromere regions and repressed nucleosome formation in the promoter regions of protein-coding genes to mediate transcriptional regulation. Based on these analyses, we integrated all twelve structural features in a model to predict more accurately nucleosome occupancy in vivo than the existing methods that mainly depend on sequence compositional features. Furthermore, we developed a novel approach, named DLaNe, that located nucleosomes by detecting peaks of structural profiles, and built a meta predictor to integrate information from different structural features. As a comparison, we also constructed a hidden Markov model (HMM to locate nucleosomes based on the profiles of these structural features. The result showed that the meta DLaNe and HMM-based method performed better than the existing methods, demonstrating the power of these structural features in predicting nucleosome positions. Conclusions Our analysis revealed that DNA structures significantly contribute to nucleosome organization and influence

  14. Interactions between isolated nucleosome core particles: A tail-bridging effect?

    Science.gov (United States)

    Mangenot, S.; Raspaud, E.; Tribet, C.; Belloni, L.; Livolant, F.

    2002-03-01

    Interactions between isolated nucleosome core particles are studied as a function of the monovalent salt concentration by osmometry and by electrophoretic mobility measurements. The data are compared to the measurements performed on the protein-free DNA fragments and also analysed using the conventional theoretical approach. At low salt, an electrostatic screening effect accounts for the variation of the second virial coefficient whereas the simple hard-core contribution becomes predominant at high salt. In the intermediate range, an attraction occurs. In the light of previous results (Mangenot et al. , Biophys. J. 82, 345 (2002)), we show that the flexible basic proteic tails are responsible for this attraction. A tail-bridging effect is discussed.

  15. Using DNA mechanics to predict intrinsic and extrinsic nucleosome positioning signals

    Science.gov (United States)

    Morozov, Alexandre

    2008-03-01

    In eukaryotic genomes, nucleosomes function to compact DNA and to regulate access to it both by simple physical occlusion and by providing the substrate for numerous covalent epigenetic tags. While nucleosome positions in vitro are determined by sequence alone, in vivo competition with other DNA-binding factors and action of chromatin remodeling enzymes play a role that needs to be quantified. We developed a biophysical, DNA mechanics-based model for the sequence dependence of DNA bending energies, and validated it against a collection of in vitro free energies of nucleosome formation and a nucleosome crystal structure; we also successfully designed both strong and poor histone binding sequences ab initio. For in vivo data from S.cerevisiae, the strongest positioning signal came from the competition with other factors rather than intrinsic nucleosome sequence preferences. Based on sequence alone, our model predicts that functional transcription factor binding sites tend to be covered by nucleosomes, yet are uncovered in vivo because functional sites cluster within a single nucleosome footprint and thus make transcription factors bind cooperatively. Similarly a weak enhancement of nucleosome binding in the TATA region becomes a strong depletion when the TATA-binding protein is included, in quantitative agreement with experiment. Our model distinguishes multiple ways in which genomic sequence influences nucleosome positions, and thus provides alternative explanations for several genome-wide experimental findings. In the future our approach will be used to rationally alter gene expression levels in model systems through redesign of nucleosome occupancy profiles.

  16. An in vitro-identified high-affinity nucleosome-positioning signal is capable of transiently positioning a nucleosome in vivo

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    Gracey Lia E

    2010-07-01

    Full Text Available Abstract Background The physiological function of eukaryotic DNA occurs in the context of nucleosomal arrays that can expose or obscure defined segments of the genome. Certain DNA sequences are capable of strongly positioning a nucleosome in vitro, suggesting the possibility that favorable intrinsic signals might reproducibly structure chromatin segments. As high-throughput sequencing analyses of nucleosome coverage in vitro and in vivo have become possible, a vigorous debate has arisen over the degree to which intrinsic DNA:nucleosome affinities orchestrate the in vivo positions of nucleosomes, thereby controlling physical accessibility of specific sequences in DNA. Results We describe here the in vivo consequences of placing a synthetic high-affinity nucleosome-positioning signal, the 601 sequence, into a DNA plasmid vector in mice. Strikingly, the 601 sequence was sufficient to position nucleosomes during an early phase after introduction of the DNA into the mice (when the plasmid vector transgene was active. This positioning capability was transient, with a loss of strong positioning at a later time point when the transgenes had become silent. Conclusions These results demonstrate an ability of DNA sequences selected solely for nucleosome affinity to organize chromatin in vivo, and the ability of other mechanisms to overcome these interactions in a dynamic nuclear environment.

  17. An advanced coarse-grained nucleosome core particle model for computer simulations of nucleosome-nucleosome interactions under varying ionic conditions.

    Directory of Open Access Journals (Sweden)

    Yanping Fan

    Full Text Available In the eukaryotic cell nucleus, DNA exists as chromatin, a compact but dynamic complex with histone proteins. The first level of DNA organization is the linear array of nucleosome core particles (NCPs. The NCP is a well-defined complex of 147 bp DNA with an octamer of histones. Interactions between NCPs are of paramount importance for higher levels of chromatin compaction. The polyelectrolyte nature of the NCP implies that nucleosome-nucleosome interactions must exhibit a great influence from both the ionic environment as well as the positively charged and highly flexible N-terminal histone tails, protruding out from the NCP. The large size of the system precludes a modelling analysis of chromatin at an all-atom level and calls for coarse-grained approximations. Here, a model of the NCP that include the globular histone core and the flexible histone tails described by one particle per each amino acid and taking into account their net charge is proposed. DNA wrapped around the histone core was approximated at the level of two base pairs represented by one bead (bases and sugar plus four beads of charged phosphate groups. Computer simulations, using a Langevin thermostat, in a dielectric continuum with explicit monovalent (K(+, divalent (Mg(2+ or trivalent (Co(NH(3(6 (3+ cations were performed for systems with one or ten NCPs. Increase of the counterion charge results in a switch from repulsive NCP-NCP interaction in the presence of K(+, to partial aggregation with Mg(2+ and to strong mutual attraction of all 10 NCPs in the presence of CoHex(3+. The new model reproduced experimental results and the structure of the NCP-NCP contacts is in agreement with available data. Cation screening, ion-ion correlations and tail bridging contribute to the NCP-NCP attraction and the new NCP model accounts for these interactions.

  18. [Structure and dynamic of the nucleosome core particle].

    Science.gov (United States)

    Bertin, Aurélie; Mangenot, Stéphanie

    2008-01-01

    In eukaryotic cell, a few meters of DNA are compacted in nuclear compartment of a few microns. This high level of compaction is an important way to regulate gene expression. In the present paper, we present a description of the organization of DNA into its first level of compaction: the nucleosome core particle. The structure of the nucleosome has been described at an atomic resolution more than 10 years ago, where DNA is wrapped around an octamer of histones. Post-translational modifications affecting histone tails have been shown to regulate the chromatin degree of compaction and thus the gene expression and regulation. The structure of the NCP is far from being frozen and is highly dynamic. Remodeling factors can induce DNA sliding around the histones, DNA transaction processes such as transcription and replication.

  19. BRCA 1-Mediated Histone Monoubiquitylation: Effect on Nucleosome Dynamics

    Science.gov (United States)

    2008-02-01

    nucleosome positioning sequence from sea urchin 5s rRNA gene; perform and analyze the ubiquitylation reaction (month 4). Done Task 2: Use single... sea urchin 5S rDNA, and oligonucleosomes were reconstituted on the same sequence repeated in tandem (208-12) (Simpson et al., 1985). Isolation of...Center, Columbus, OH 43210), who provided BRCA1/BARD1 protein complex. b. Acquire and perform quality tests on the other components of the

  20. Translation efficiency in yeasts correlates with nucleosome formation in promoters.

    Science.gov (United States)

    Matushkin, Yu G; Levitsky, V G; Orlov, Yu L; Likhoshvai, V A; Kolchanov, N A

    2013-01-01

    Elongation efficiency index (EEI) was suggested earlier to estimate gene expression efficiency by nucleotide context of coding sequence in unicellular organisms. We have analyzed association between EEI and nucleosome formation potential (NFP) in 5' regulatory regions upstream translation initiation site (TIS) from two yeast species. Theoretical estimations of NFP based on DNA sequence were obtained by Recon method. Experimental estimation of nucleosome occupancy was obtained by high-throughput sequencing data of nucleosomal DNA in Saccharomyces cerevisiae . For the sample of all genes correlation coefficient was calculated between two vectors: vector of NFP values for fixed position relative to TIS and vector of EEI values. Profiles of correlation coefficients of NFP and EEI were counted in (-600; +600) regions relative to TIS for gene sequences extracted from GenBank. We found regions of strong negative dependence between NFP and EEI for all genes as well as for 10% highly expressed genes in Schizosaccharomyces pombe (10% of EEI-highest genes). At the same time, we found positive dependence between NFP and EEI for all genes and for low expressed genes in S. cerevisiae (10% of EEI-lowest genes). The association between NFP and EEI could be explained by evolutionary selection of context characteristics of nucleotide sequences for gene expression optimization.

  1. Cracking the chromatin code: Precise rule of nucleosome positioning

    Science.gov (United States)

    Trifonov, Edward N.

    2011-03-01

    Various aspects of packaging DNA in eukaryotic cells are outlined in physical rather than biological terms. The informational and physical nature of packaging instructions encoded in DNA sequences is discussed with the emphasis on signal processing difficulties - very low signal-to-noise ratio and high degeneracy of the nucleosome positioning signal. As the author has been contributing to the field from its very onset in 1980, the review is mostly focused at the works of the author and his colleagues. The leading concept of the overview is the role of deformational properties of DNA in the nucleosome positioning. The target of the studies is to derive the DNA bendability matrix describing where along the DNA various dinucleotide elements should be positioned, to facilitate its bending in the nucleosome. Three different approaches are described leading to derivation of the DNA deformability sequence pattern, which is a simplified linear presentation of the bendability matrix. All three approaches converge to the same unique sequence motif CGRAAATTTYCG or, in binary form, YRRRRRYYYYYR, both representing the chromatin code.

  2. A role for tuned levels of nucleosome remodeler subunit ACF1 during Drosophila oogenesis.

    Science.gov (United States)

    Börner, Kenneth; Jain, Dhawal; Vazquez-Pianzola, Paula; Vengadasalam, Sandra; Steffen, Natascha; Fyodorov, Dmitry V; Tomancak, Pavel; Konev, Alexander; Suter, Beat; Becker, Peter B

    2016-03-15

    The Chromatin Accessibility Complex (CHRAC) consists of the ATPase ISWI, the large ACF1 subunit and a pair of small histone-like proteins, CHRAC-14/16. CHRAC is a prototypical nucleosome sliding factor that mobilizes nucleosomes to improve the regularity and integrity of the chromatin fiber. This may facilitate the formation of repressive chromatin. Expression of the signature subunit ACF1 is restricted during embryonic development, but remains high in primordial germ cells. Therefore, we explored roles for ACF1 during Drosophila oogenesis. ACF1 is expressed in somatic and germline cells, with notable enrichment in germline stem cells and oocytes. The asymmetrical localization of ACF1 to these cells depends on the transport of the Acf1 mRNA by the Bicaudal-D/Egalitarian complex. Loss of ACF1 function in the novel Acf1(7) allele leads to defective egg chambers and their elimination through apoptosis. In addition, we find a variety of unusual 16-cell cyst packaging phenotypes in the previously known Acf1(1) allele, with a striking prevalence of egg chambers with two functional oocytes at opposite poles. Surprisingly, we found that the Acf1(1) deletion--despite disruption of the Acf1 reading frame--expresses low levels of a PHD-bromodomain module from the C-terminus of ACF1 that becomes enriched in oocytes. Expression of this module from the Acf1 genomic locus leads to packaging defects in the absence of functional ACF1, suggesting competitive interactions with unknown target molecules. Remarkably, a two-fold overexpression of CHRAC (ACF1 and CHRAC-16) leads to increased apoptosis and packaging defects. Evidently, finely tuned CHRAC levels are required for proper oogenesis. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. The nucleosome landscape of Plasmodium falciparum reveals chromatin architecture and dynamics of regulatory sequences

    Science.gov (United States)

    Kensche, Philip Reiner; Hoeijmakers, Wieteke Anna Maria; Toenhake, Christa Geeke; Bras, Maaike; Chappell, Lia; Berriman, Matthew; Bártfai, Richárd

    2016-01-01

    In eukaryotes, the chromatin architecture has a pivotal role in regulating all DNA-associated processes and it is central to the control of gene expression. For Plasmodium falciparum, a causative agent of human malaria, the nucleosome positioning profile of regulatory regions deserves particular attention because of their extreme AT-content. With the aid of a highly controlled MNase-seq procedure we reveal how positioning of nucleosomes provides a structural and regulatory framework to the transcriptional unit by demarcating landmark sites (transcription/translation start and end sites). In addition, our analysis provides strong indications for the function of positioned nucleosomes in splice site recognition. Transcription start sites (TSSs) are bordered by a small nucleosome-depleted region, but lack the stereotypic downstream nucleosome arrays, highlighting a key difference in chromatin organization compared to model organisms. Furthermore, we observe transcription-coupled eviction of nucleosomes on strong TSSs during intraerythrocytic development and demonstrate that nucleosome positioning and dynamics can be predictive for the functionality of regulatory DNA elements. Collectively, the strong nucleosome positioning over splice sites and surrounding putative transcription factor binding sites highlights the regulatory capacity of the nucleosome landscape in this deadly human pathogen. PMID:26578577

  4. A multi-layer method to study genome-scale positions of nucleosomes.

    Science.gov (United States)

    Di Gesù, Vito; Lo Bosco, Giosuè; Pinello, Luca; Yuan, Guo-Cheng; Corona, Davide F V

    2009-02-01

    The basic unit of eukaryotic chromatin is the nucleosome, consisting of about 150 bp of DNA wrapped around a protein core made of histone proteins. Nucleosomes position is modulated in vivo to regulate fundamental nuclear processes. To measure nucleosome positions on a genomic scale both theoretical and experimental approaches have been recently reported. We have developed a new method, Multi-Layer Model (MLM), for the analysis of nucleosome position data obtained with microarray-based approach. The MLM is a feature extraction method in which the input data is processed by a classifier to distinguish between several kinds of patterns. We applied our method to simulated-synthetic and experimental nucleosome position data and found that besides a high nucleosome recognition and a strong agreement with standard statistical methods, the MLM can identify distinct classes of nucleosomes, making it an important tool for the genome wide analysis of nucleosome position and function. In conclusion, the MLM allows a better representation of nucleosome position data and a significant reduction in computational time.

  5. H4 Tails Potentially Produce the Diversity in the Orientation of Two Nucleosomes.

    Science.gov (United States)

    Ishida, Hisashi; Kono, Hidetoshi

    2017-09-05

    Histone tails play an important role in internucleosomal interaction and chromatin compaction. To understand how the H4 tails are involved in the internucleosomal interaction, an adaptively biased molecular dynamics simulation of 63 models of two stacked nucleosomes, each with the H4 tails in different locations, was carried out. This simulation generated a variety of orientations of the separated nucleosomes depending on the formation of the H4 tail bridge between the H4 tails and the DNA of the neighboring nucleosomes. For the models that showed distinctive orientations of the two nucleosomes, the free energies of the separation of the nucleosomes were further investigated using umbrella sampling simulations. The attractive force between the nucleosomes was estimated from the free energies; the force when two H4 tail bridges formed varied from 36 to 63 pN, depending on the formation of the H4 tail-bridge and the interfacial interaction, whereas the force reduced to 15-18 pN after either one of the H4 tail bridges had broken, regardless of the conformation of the H4 tail. Additional simulations of the nucleosomes show that when the H4 tail was truncated, the force between the nucleosomes became repulsive (from-3 to -7 pN). We concluded that the H4 tails potentially produce the diversity in the orientation of the two nucleosomes, which would contribute to the polymorphism of the chromatin structure. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. The nucleosome landscape of Plasmodium falciparum reveals chromatin architecture and dynamics of regulatory sequences.

    Science.gov (United States)

    Kensche, Philip Reiner; Hoeijmakers, Wieteke Anna Maria; Toenhake, Christa Geeke; Bras, Maaike; Chappell, Lia; Berriman, Matthew; Bártfai, Richárd

    2016-03-18

    In eukaryotes, the chromatin architecture has a pivotal role in regulating all DNA-associated processes and it is central to the control of gene expression. For Plasmodium falciparum, a causative agent of human malaria, the nucleosome positioning profile of regulatory regions deserves particular attention because of their extreme AT-content. With the aid of a highly controlled MNase-seq procedure we reveal how positioning of nucleosomes provides a structural and regulatory framework to the transcriptional unit by demarcating landmark sites (transcription/translation start and end sites). In addition, our analysis provides strong indications for the function of positioned nucleosomes in splice site recognition. Transcription start sites (TSSs) are bordered by a small nucleosome-depleted region, but lack the stereotypic downstream nucleosome arrays, highlighting a key difference in chromatin organization compared to model organisms. Furthermore, we observe transcription-coupled eviction of nucleosomes on strong TSSs during intraerythrocytic development and demonstrate that nucleosome positioning and dynamics can be predictive for the functionality of regulatory DNA elements. Collectively, the strong nucleosome positioning over splice sites and surrounding putative transcription factor binding sites highlights the regulatory capacity of the nucleosome landscape in this deadly human pathogen. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  7. The role of histone H4 biotinylation in the structure of nucleosomes.

    Directory of Open Access Journals (Sweden)

    Nina A Filenko

    2011-01-01

    Full Text Available Post-translational modifications of histones play important roles in regulating nucleosome structure and gene transcription. It has been shown that biotinylation of histone H4 at lysine-12 in histone H4 (K12Bio-H4 is associated with repression of a number of genes. We hypothesized that biotinylation modifies the physical structure of nucleosomes, and that biotin-induced conformational changes contribute to gene silencing associated with histone biotinylation.To test this hypothesis we used atomic force microscopy to directly analyze structures of nucleosomes formed with biotin-modified and non-modified H4. The analysis of the AFM images revealed a 13% increase in the length of DNA wrapped around the histone core in nucleosomes with biotinylated H4. This statistically significant (p<0.001 difference between native and biotinylated nucleosomes corresponds to adding approximately 20 bp to the classical 147 bp length of nucleosomal DNA.The increase in nucleosomal DNA length is predicted to stabilize the association of DNA with histones and therefore to prevent nucleosomes from unwrapping. This provides a mechanistic explanation for the gene silencing associated with K12Bio-H4. The proposed single-molecule AFM approach will be instrumental for studying the effects of various epigenetic modifications of nucleosomes, in addition to biotinylation.

  8. Stable complex formation of CENP-B with the CENP-A nucleosome

    Science.gov (United States)

    Fujita, Risa; Otake, Koichiro; Arimura, Yasuhiro; Horikoshi, Naoki; Miya, Yuta; Shiga, Tatsuya; Osakabe, Akihisa; Tachiwana, Hiroaki; Ohzeki, Jun-ichirou; Larionov, Vladimir; Masumoto, Hiroshi; Kurumizaka, Hitoshi

    2015-01-01

    CENP-A and CENP-B are major components of centromeric chromatin. CENP-A is the histone H3 variant, which forms the centromere-specific nucleosome. CENP-B specifically binds to the CENP-B box DNA sequence on the centromere-specific repetitive DNA. In the present study, we found that the CENP-A nucleosome more stably retains human CENP-B than the H3.1 nucleosome in vitro. Specifically, CENP-B forms a stable complex with the CENP-A nucleosome, when the CENP-B box sequence is located at the proximal edge of the nucleosome. Surprisingly, the CENP-B binding was weaker when the CENP-B box sequence was located in the distal linker region of the nucleosome. This difference in CENP-B binding, depending on the CENP-B box location, was not observed with the H3.1 nucleosome. Consistently, we found that the DNA-binding domain of CENP-B specifically interacted with the CENP-A-H4 complex, but not with the H3.1-H4 complex, in vitro. These results suggested that CENP-B forms a more stable complex with the CENP-A nucleosome through specific interactions with CENP-A, if the CENP-B box is located proximal to the CENP-A nucleosome. Our in vivo assay also revealed that CENP-B binding in the vicinity of the CENP-A nucleosome substantially stabilizes the CENP-A nucleosome on alphoid DNA in human cells. PMID:25916850

  9. Linker histone H1 and H3K56 acetylation are antagonistic regulators of nucleosome dynamics.

    Science.gov (United States)

    Bernier, Morgan; Luo, Yi; Nwokelo, Kingsley C; Goodwin, Michelle; Dreher, Sarah J; Zhang, Pei; Parthun, Mark R; Fondufe-Mittendorf, Yvonne; Ottesen, Jennifer J; Poirier, Michael G

    2015-12-09

    H1 linker histones are highly abundant proteins that compact nucleosomes and chromatin to regulate DNA accessibility and transcription. However, the mechanisms that target H1 regulation to specific regions of eukaryotic genomes are unknown. Here we report fluorescence measurements of human H1 regulation of nucleosome dynamics and transcription factor (TF) binding within nucleosomes. H1 does not block TF binding, instead it suppresses nucleosome unwrapping to reduce DNA accessibility within H1-bound nucleosomes. We then investigated H1 regulation by H3K56 and H3K122 acetylation, two transcriptional activating histone post translational modifications (PTMs). Only H3K56 acetylation, which increases nucleosome unwrapping, abolishes H1.0 reduction of TF binding. These findings show that nucleosomes remain dynamic, while H1 is bound and H1 dissociation is not required for TF binding within the nucleosome. Furthermore, our H3K56 acetylation measurements suggest that a single-histone PTM can define regions of the genome that are not regulated by H1.

  10. Nucleosomes are context-specific, H2A.Z-modulated barriers to RNA polymerase.

    Science.gov (United States)

    Weber, Christopher M; Ramachandran, Srinivas; Henikoff, Steven

    2014-03-06

    Nucleosomes are barriers to transcription in vitro; however, their effects on RNA polymerase in vivo are unknown. Here we describe a simple and general strategy to comprehensively map the positions of elongating and arrested RNA polymerase II (RNAPII) at nucleotide resolution. We find that the entry site of the first (+1) nucleosome is a barrier to RNAPII for essentially all genes, including those undergoing regulated pausing farther upstream. In contrast to the +1 nucleosome, gene body nucleosomes are low barriers and cause RNAPII stalling both at the entry site and near the dyad axis. The extent of the +1 nucleosome barrier correlates with nucleosome occupancy but anticorrelates with enrichment of histone variant H2A.Z. Importantly, depletion of H2A.Z from a nucleosome position results in a higher barrier to RNAPII. Our results suggest that nucleosomes present significant, context-specific barriers to RNAPII in vivo that can be tuned by the incorporation of H2A.Z. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Asymmetric breathing motions of nucleosomal DNA and the role of histone tails.

    Science.gov (United States)

    Chakraborty, Kaushik; Loverde, Sharon M

    2017-08-14

    The most important packing unit of DNA in the eukaryotic cell is the nucleosome. It undergoes large-scale structural re-arrangements during different cell cycles. For example, the disassembly of the nucleosome is one of the key steps for DNA replication, whereas reassembly occurs after replication. Thus, conformational dynamics of the nucleosome is crucial for different DNA metabolic processes. We perform three different sets of atomistic molecular dynamics simulations of the nucleosome core particle at varying degrees of salt conditions for a total of 0.7 μs simulation time. We find that the conformational dynamics of the nucleosomal DNA tails are oppositely correlated from each other during the initial breathing motions. Furthermore, the strength of the interaction of the nucleosomal DNA tail with the neighboring H2A histone tail modulates the conformational state of the nucleosomal DNA tail. With increasing salt concentration, the degree of asymmetry in the conformation of the nucleosomal DNA tails decreases as both tails tend to unwrap. This direct correlation between the asymmetric breathing motions of the DNA tails and the H2A histone tails, and its decrease at higher salt concentrations, may play a significant role in the molecular pathway of unwrapping.

  12. Contributions of histone H3 nucleosome core surface mutations to chromatin structures, silencing and DNA repair.

    Directory of Open Access Journals (Sweden)

    Michel Fink

    Full Text Available Histone H3 mutations in residues that cluster in a discrete region on the nucleosome surface around lysine 79 of H3 affect H3-K79 methylation, impair transcriptional silencing in subtelomeric chromatin, and reveal distinct contributions of histone H3 to various DNA-damage response and repair pathways. These residues might act by recruitment of silencing and DNA-damage response factors. Alternatively, their location on the nucleosome surface suggests a possible involvement in nucleosome positioning, stability and nucleosome interactions. Here, we show that the yeast H3 mutants hht2-T80A, hht2-K79E, hht2-L70S, and hht2-E73D show normal nucleosome positioning and stability in minichromosomes. However, loss of silencing in a subtelomeric URA3 gene correlates with a shift of the promoter nucleosome, while nucleosome positions and stability in the coding region are maintained. Moreover, the H3 mutants show normal repair of UV lesions by photolyase and nucleotide excision repair in minichromosomes and slightly enhanced repair in the subtelomeric region. Thus, these results support a role of those residues in the recruitment of silencing proteins and argue against a general role in nucleosome organization.

  13. Differential Nucleosome Occupancies across Oct4-Sox2 Binding Sites in Murine Embryonic Stem Cells.

    Directory of Open Access Journals (Sweden)

    Amy Sebeson

    Full Text Available The binding sequence for any transcription factor can be found millions of times within a genome, yet only a small fraction of these sequences encode functional transcription factor binding sites. One of the reasons for this dichotomy is that many other factors, such as nucleosomes, compete for binding. To study how the competition between nucleosomes and transcription factors helps determine a functional transcription factor site from a predicted transcription factor site, we compared experimentally-generated in vitro nucleosome occupancy with in vivo nucleosome occupancy and transcription factor binding in murine embryonic stem cells. Using a solution hybridization enrichment technique, we generated a high-resolution nucleosome map from targeted regions of the genome containing predicted sites and functional sites of Oct4/Sox2 regulation. We found that at Pax6 and Nes, which are bivalently poised in stem cells, functional Oct4 and Sox2 sites show high amounts of in vivo nucleosome displacement compared to in vitro. Oct4 and Sox2, which are active, show no significant displacement of in vivo nucleosomes at functional sites, similar to nonfunctional Oct4/Sox2 binding. This study highlights a complex interplay between Oct4 and Sox2 transcription factors and nucleosomes among different target genes, which may result in distinct patterns of stem cell gene regulation.

  14. Asymmetric breathing motions of nucleosomal DNA and the role of histone tails

    Science.gov (United States)

    Chakraborty, Kaushik; Loverde, Sharon M.

    2017-08-01

    The most important packing unit of DNA in the eukaryotic cell is the nucleosome. It undergoes large-scale structural re-arrangements during different cell cycles. For example, the disassembly of the nucleosome is one of the key steps for DNA replication, whereas reassembly occurs after replication. Thus, conformational dynamics of the nucleosome is crucial for different DNA metabolic processes. We perform three different sets of atomistic molecular dynamics simulations of the nucleosome core particle at varying degrees of salt conditions for a total of 0.7 μs simulation time. We find that the conformational dynamics of the nucleosomal DNA tails are oppositely correlated from each other during the initial breathing motions. Furthermore, the strength of the interaction of the nucleosomal DNA tail with the neighboring H2A histone tail modulates the conformational state of the nucleosomal DNA tail. With increasing salt concentration, the degree of asymmetry in the conformation of the nucleosomal DNA tails decreases as both tails tend to unwrap. This direct correlation between the asymmetric breathing motions of the DNA tails and the H2A histone tails, and its decrease at higher salt concentrations, may play a significant role in the molecular pathway of unwrapping.

  15. Dynamic Nucleosome Movement Provides Structural Information of Topological Chromatin Domains in Living Human Cells.

    Science.gov (United States)

    Shinkai, Soya; Nozaki, Tadasu; Maeshima, Kazuhiro; Togashi, Yuichi

    2016-10-01

    The mammalian genome is organized into submegabase-sized chromatin domains (CDs) including topologically associating domains, which have been identified using chromosome conformation capture-based methods. Single-nucleosome imaging in living mammalian cells has revealed subdiffusively dynamic nucleosome movement. It is unclear how single nucleosomes within CDs fluctuate and how the CD structure reflects the nucleosome movement. Here, we present a polymer model wherein CDs are characterized by fractal dimensions and the nucleosome fibers fluctuate in a viscoelastic medium with memory. We analytically show that the mean-squared displacement (MSD) of nucleosome fluctuations within CDs is subdiffusive. The diffusion coefficient and the subdiffusive exponent depend on the structural information of CDs. This analytical result enabled us to extract information from the single-nucleosome imaging data for HeLa cells. Our observation that the MSD is lower at the nuclear periphery region than the interior region indicates that CDs in the heterochromatin-rich nuclear periphery region are more compact than those in the euchromatin-rich interior region with respect to the fractal dimensions as well as the size. Finally, we evaluated that the average size of CDs is in the range of 100-500 nm and that the relaxation time of nucleosome movement within CDs is a few seconds. Our results provide physical and dynamic insights into the genome architecture in living cells.

  16. Nanopores suggest a negligible influence of CpG methylation on nucleosome packaging and stability.

    Science.gov (United States)

    Langecker, Martin; Ivankin, Andrey; Carson, Spencer; Kinney, Shannon R M; Simmel, Friedrich C; Wanunu, Meni

    2015-01-14

    Nucleosomes are the fundamental repeating units of chromatin, and dynamic regulation of their positioning along DNA governs gene accessibility in eukaryotes. Although epigenetic factors have been shown to influence nucleosome structure and dynamics, the impact of DNA methylation on nucleosome packaging remains controversial. Further, all measurements to date have been carried out under zero-force conditions. In this paper, we present the first automated force measurements that probe the impact of CpG DNA methylation on nucleosome stability. In solid-state nanopore force spectroscopy, a nucleosomal DNA tail is captured into a pore and pulled on with a time-varying electrophoretic force until unraveling is detected. This is automatically repeated for hundreds of nucleosomes, yielding statistics of nucleosome lifetime vs electrophoretic force. The force geometry, which is similar to displacement forces exerted by DNA polymerases and helicases, reveals that nucleosome stability is sensitive to DNA sequence yet insensitive to CpG methylation. Our label-free method provides high-throughput data that favorably compares with other force spectroscopy experiments and is suitable for studying a variety of DNA-protein complexes.

  17. Characterization of the Nucleosome Landscape by Micrococcal Nuclease-Sequencing (MNase-seq).

    Science.gov (United States)

    Hoeijmakers, Wieteke Anna Maria; Bártfai, Richárd

    2018-01-01

    MNase-seq allows the genome-wide examination of the nucleosome landscape by determination of nucleosome positioning and occupancy. Typically, native or formaldehyde fixed chromatin is subjected to digestion by micrococcal nuclease (MNase), which degrades linker DNA and yields mainly mono-nucleosomes. The resulting material can be processed directly or can be subjected to an optional chromatin immunoprecipitation step (MNase-ChIP-seq). De-crosslinked and purified DNA is then subjected to next-generation sequencing. The protocol presented here has been tailored for the analysis of nucleosome landscape in the malaria parasite, Plasmodium falciparum, but most steps are directly applicable to other cell types. We also discuss general considerations for experimental design and computational analysis, which are crucial for accurate investigation of the nucleosome landscape.

  18. No need for a power stroke in ISWI-mediated nucleosome sliding

    Science.gov (United States)

    Ludwigsen, Johanna; Klinker, Henrike; Mueller-Planitz, Felix

    2013-01-01

    Nucleosome remodelling enzymes of the ISWI family reposition nucleosomes in eukaryotes. ISWI contains an ATPase and a HAND-SANT-SLIDE (HSS) domain. Conformational changes between these domains have been proposed to be critical for nucleosome repositioning by pulling flanking DNA into the nucleosome. We inserted flexible linkers at strategic sites in ISWI to disrupt this putative power stroke and assess its functional importance by quantitative biochemical assays. Notably, the flexible linkers did not disrupt catalysis. Instead of engaging in a power stroke, the HSS module might therefore assist DNA to ratchet into the nucleosome. Our results clarify the roles had by the domains and suggest that the HSS domain evolved to optimize a rudimentary remodelling engine. PMID:24113208

  19. Stimulation of the Drosophila immune system alters genome-wide nucleosome occupancy

    Directory of Open Access Journals (Sweden)

    Yingxue Ren

    2015-03-01

    Full Text Available In eukaryotes, nucleosomes participate in all DNA-templated events by regulating access to the underlying DNA sequence. However, nucleosome dynamics during a genome response have not been well characterized [1,2]. We stimulated Drosophila S2 cells with heat-killed Gram-negative bacteria Salmonella typhimurium, and mapped genome-wide nucleosome occupancy at high temporal resolution by MNase-seq using Illumina HiSeq 2500. We show widespread nucleosome occupancy change in S2 cells during the immune response, with the significant nucleosomal loss occurring at 4 h after stimulation. Data have been deposited to the Gene Expression Omnibus (GEO database repository with the dataset identifier GSE64507.

  20. RPA binds histone H3-H4 and functions in DNA replication-coupled nucleosome assembly.

    Science.gov (United States)

    Liu, Shaofeng; Xu, Zhiyun; Leng, He; Zheng, Pu; Yang, Jiayi; Chen, Kaifu; Feng, Jianxun; Li, Qing

    2017-01-27

    DNA replication-coupled nucleosome assembly is essential to maintain genome integrity and retain epigenetic information. Multiple involved histone chaperones have been identified, but how nucleosome assembly is coupled to DNA replication remains elusive. Here we show that replication protein A (RPA), an essential replisome component that binds single-stranded DNA, has a role in replication-coupled nucleosome assembly. RPA directly binds free H3-H4. Assays using a synthetic sequence that mimics freshly unwound single-stranded DNA at replication fork showed that RPA promotes DNA-(H3-H4) complex formation immediately adjacent to double-stranded DNA. Further, an RPA mutant defective in H3-H4 binding exhibited attenuated nucleosome assembly on nascent chromatin. Thus, we propose that RPA functions as a platform for targeting histone deposition to replication fork, through which RPA couples nucleosome assembly with ongoing DNA replication. Copyright © 2017, American Association for the Advancement of Science.

  1. Naturally occuring nucleosome positioning signals in human exons and introns

    DEFF Research Database (Denmark)

    Baldi, Pierre; Brunak, Søren; Chauvin, Yves

    1996-01-01

    We describe the structural implications of a periodic pattern found in human exons and introns by hidden Markov models. We show that exons (besides the reading frame) have a specific sequential structure in the form of a pattern with triplet consensus non-T(A/T)G, and a minimal periodicity...... faces inward and is compressed. The in-phase triplets are located adjacent to GCC/GGC triplets known to have the strongest bias in their positioning on the nucleosome. Analysis of mRNA sequences encoding proteins with known tertiary structure exclude the possibility that the pattern is a consequence...

  2. Distinct influences of tandem repeats and retrotransposons on CENH3 nucleosome positioning

    Directory of Open Access Journals (Sweden)

    Gent Jonathan I

    2011-02-01

    Full Text Available Abstract Background Unique structural characteristics of centromere chromatin enable it to support assembly of the kinetochore and its associated tensions. The histone H3 variant CENH3 (centromeric histone H3 is viewed as the key element of centromere chromatin and its interaction with centromere DNA is epigenetic in that its localization to centromeres is not sequence-dependent. Results In order to investigate what influence the DNA sequence exerts on CENH3 chromatin structure, we examined CENH3 nucleosome footprints on maize centromere DNA. We found a predominant average nucleosome spacing pattern of roughly 190-bp intervals, which was also the dominant arrangement for nucleosomes genome-wide. For CENH3-containing nucleosomes, distinct modes of nucleosome positioning were evident within that general spacing constraint. Over arrays of the major ~156-bp centromeric satellite sequence (tandem repeat CentC, nucleosomes were not positioned in register with CentC monomers but in conformity with a striking ~10-bp periodicity of AA/TT dimers within the sequence. In contrast, nucleosomes on a class of centromeric retrotransposon (CRM2 lacked a detectable AA/TT periodicity but exhibited tightly phased positioning. Conclusions These data support a model in which general chromatin factors independent of both DNA sequence and CENH3 enforce roughly uniform centromeric nucleosome spacing while allowing flexibility in the mode in which nucleosomes are positioned. In the case of tandem repeat DNA, the natural bending effects related to AA/TT periodicity produce an energetically-favourable arrangement consistent with conformationally rigid nucleosomes and stable chromatin at centromeres.

  3. In silico evidence for sequence-dependent nucleosome sliding

    Energy Technology Data Exchange (ETDEWEB)

    Lequieu, Joshua; Schwartz, David C.; de Pablo, Juan J.

    2017-10-18

    Nucleosomes represent the basic building block of chromatin and provide an important mechanism by which cellular processes are controlled. The locations of nucleosomes across the genome are not random but instead depend on both the underlying DNA sequence and the dynamic action of other proteins within the nucleus. These processes are central to cellular function, and the molecular details of the interplay between DNA sequence and nudeosome dynamics remain poorly understood. In this work, we investigate this interplay in detail by relying on a molecular model, which permits development of a comprehensive picture of the underlying free energy surfaces and the corresponding dynamics of nudeosome repositioning. The mechanism of nudeosome repositioning is shown to be strongly linked to DNA sequence and directly related to the binding energy of a given DNA sequence to the histone core. It is also demonstrated that chromatin remodelers can override DNA-sequence preferences by exerting torque, and the histone H4 tail is then identified as a key component by which DNA-sequence, histone modifications, and chromatin remodelers could in fact be coupled.

  4. FSAP-mediated nucleosome release from late apoptotic cells is inhibited by autoantibodies present in SLE.

    Science.gov (United States)

    Marsman, Gerben; Stephan, Femke; de Leeuw, Karina; Bulder, Ingrid; Ruinard, Jessica T; de Jong, Jan; Westra, Johanna; Bultink, Irene E M; Voskuyl, Alexandre E; Aarden, Lucien A; Luken, Brenda M; Kallenberg, Cees G M; Zeerleder, Sacha

    2016-03-01

    Inefficient clearance of apoptotic cells and the subsequent exposure of the immune system to nuclear contents are crucially involved in the pathogenesis of systemic lupus erythematosus (SLE). Factor VII-activating protease (FSAP) is activated in serum upon contact with dead cells, and releases nucleosomes from late apoptotic cells into the extracellular environment. We investigated whether FSAP-mediated nucleosome release from late apoptotic cells is affected in SLE patients. Nucleosome release in sera of 27 SLE patients and 30 healthy controls was investigated by incubating late apoptotic Jurkat cells with serum and analyzing the remaining DNA content by flow cytometry. We found that nucleosome release in sera of SLE patients with high disease activity was significantly decreased when compared with that in SLE sera obtained during low disease activity or from healthy individuals. Upon removal of IgG/IgM antibodies from SLE sera, nucleosome release was restored. Similarly, monoclonal antinuclear antibodies inhibited nucleosome release in healthy donor serum or by plasma-purified FSAP. This inhibition was lost when Fab fragments were used, suggesting that antigen cross-linking is involved. In conclusion, FSAP-mediated nucleosome release from late apoptotic cells is greatly impaired in SLE patient sera, possibly hampering the clearance of these cells and thereby propagating inflammation. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Investigation of the interaction between berberine and nucleosomes in solution: Spectroscopic and equilibrium dialysis approach

    Science.gov (United States)

    Rabbani-Chadegani, Azra; Mollaei, Hossein; Sargolzaei, Javad

    2017-02-01

    Berberine is a natural plant alkaloid with high pharmacological potential. Although its interaction with free DNA has been the subject of several reports, to date there is no work concerning the effect of berberine on nucleoprotein structure of DNA, the nucleosomes. The present study focuses on the binding affinity of berberine to nucleosomes and histone H1 employing various spectroscopic techniques, fluorescence, circular dichroism, thermal denaturation as well as equilibrium dialysis. The results showed that the binding of berberine to nucleosomes is positive cooperative with Ka = 5.57 × 103 M- 1. Berberine quenched with the chromophores of protein moiety of nucleosomes and reduced fluorescence emission intensity at 335 nm with Ksv value of 0.135. Binding of berberine to nucleosomes decreased the absorbance at 210 and 260 nm, produced hypochromicity in thermal denaturation profiles and its affinity to nucleoprotein structure of nucleosomes was much higher than to free DNA. Berberine also exhibited high affinity to histone H1 in solution and the binding was positive cooperative with. Ka = 3.61 × 103 M- 1. Moreover berberine decreased fluorescence emission intensity of H1 by quenching with tyrosine residue in its globular core domain. The circular dichroism profiles demonstrated that the binding of drug induced secondary structural changes in both DNA stacking and histone H1. It is concluded that berberine is genotoxic drug, interacts with nucleosomes and in this process histone H1 is involved to exert its anticancer activity.

  6. Unbiased chromatin accessibility profiling by RED-seq uncovers unique features of nucleosome variants in vivo.

    Science.gov (United States)

    Chen, Poshen B; Zhu, Lihua J; Hainer, Sarah J; McCannell, Kurtis N; Fazzio, Thomas G

    2014-12-15

    Differential accessibility of DNA to nuclear proteins underlies the regulation of numerous cellular processes. Although DNA accessibility is primarily determined by the presence or absence of nucleosomes, differences in nucleosome composition or dynamics may also regulate accessibility. Methods for mapping nucleosome positions and occupancies genome-wide (MNase-seq) have uncovered the nucleosome landscapes of many different cell types and organisms. Conversely, methods specialized for the detection of large nucleosome-free regions of chromatin (DNase-seq, FAIRE-seq) have uncovered numerous gene regulatory elements. However, these methods are less successful in measuring the accessibility of DNA sequences within nucelosome arrays. Here we probe the genome-wide accessibility of multiple cell types in an unbiased manner using restriction endonuclease digestion of chromatin coupled to deep sequencing (RED-seq). Using this method, we identified differences in chromatin accessibility between populations of cells, not only in nucleosome-depleted regions of the genome (e.g., enhancers and promoters), but also within the majority of the genome that is packaged into nucleosome arrays. Furthermore, we identified both large differences in chromatin accessibility in distinct cell lineages and subtle but significant changes during differentiation of mouse embryonic stem cells (ESCs). Most significantly, using RED-seq, we identified differences in accessibility among nucleosomes harboring well-studied histone variants, and show that these differences depend on factors required for their deposition. Using an unbiased method to probe chromatin accessibility genome-wide, we uncover unique features of chromatin structure that are not observed using more widely-utilized methods. We demonstrate that different types of nucleosomes within mammalian cells exhibit different degrees of accessibility. These findings provide significant insight into the regulation of DNA accessibility.

  7. Partially Assembled Nucleosome Structures at Atomic Detail.

    Science.gov (United States)

    Rychkov, Georgy N; Ilatovskiy, Andrey V; Nazarov, Igor B; Shvetsov, Alexey V; Lebedev, Dmitry V; Konev, Alexander Y; Isaev-Ivanov, Vladimir V; Onufriev, Alexey V

    2017-02-07

    The evidence is now overwhelming that partially assembled nucleosome states (PANS) are as important as the canonical nucleosome structure for the understanding of how accessibility to genomic DNA is regulated in cells. We use a combination of molecular dynamics simulation and atomic force microscopy to deliver, in atomic detail, structural models of three key PANS: the hexasome (H2A·H2B)·(H3·H4)2, the tetrasome (H3·H4)2, and the disome (H3·H4). Despite fluctuations of the conformation of the free DNA in these structures, regions of protected DNA in close contact with the histone core remain stable, thus establishing the basis for the understanding of the role of PANS in DNA accessibility regulation. On average, the length of protected DNA in each structure is roughly 18 basepairs per histone protein. Atomistically detailed PANS are used to explain experimental observations; specifically, we discuss interpretation of atomic force microscopy, Förster resonance energy transfer, and small-angle x-ray scattering data obtained under conditions when PANS are expected to exist. Further, we suggest an alternative interpretation of a recent genome-wide study of DNA protection in active chromatin of fruit fly, leading to a conclusion that the three PANS are present in actively transcribing regions in a substantial amount. The presence of PANS may not only be a consequence, but also a prerequisite for fast transcription in vivo. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  8. The supercoiling state of DNA determines the handedness of both H3 and CENP-A nucleosomes.

    Science.gov (United States)

    Vlijm, R; Kim, S H; De Zwart, P L; Dalal, Y; Dekker, C

    2017-02-02

    Nucleosomes form the unit structure of the genome in eukaryotes, thereby constituting a fundamental tenet of chromatin biology. In canonical nucleosomes, DNA wraps around the histone octamer in a left-handed toroidal ramp. Here, in single-molecule magnetic tweezers studies of chaperone-assisted nucleosome assembly, we show that the handedness of the DNA wrapping around the nucleosome core is intrinsically ambidextrous, and depends on the pre-assembly supercoiling state of the DNA, i.e., it is not uniquely determined by the octameric histone core. Nucleosomes assembled onto negatively supercoiled DNA are found to exhibit a left-handed conformation, whereas assembly onto positively supercoiled DNA results in right-handed nucleosomes. This intrinsic flexibility to adopt both chiralities is observed both for canonical H3 nucleosomes, and for centromere-specific variant CENP-A nucleosomes. These data support recent advances suggesting an intrinsic adaptability of the nucleosome, and provide insights into how nucleosomes might rapidly re-assemble after cellular processes that generate positive supercoiling in vivo.

  9. Sequence-dependent nucleosome sliding in rotation-coupled and uncoupled modes revealed by molecular simulations.

    Science.gov (United States)

    Niina, Toru; Brandani, Giovanni B; Tan, Cheng; Takada, Shoji

    2017-12-01

    While nucleosome positioning on eukaryotic genome play important roles for genetic regulation, molecular mechanisms of nucleosome positioning and sliding along DNA are not well understood. Here we investigated thermally-activated spontaneous nucleosome sliding mechanisms developing and applying a coarse-grained molecular simulation method that incorporates both long-range electrostatic and short-range hydrogen-bond interactions between histone octamer and DNA. The simulations revealed two distinct sliding modes depending on the nucleosomal DNA sequence. A uniform DNA sequence showed frequent sliding with one base pair step in a rotation-coupled manner, akin to screw-like motions. On the contrary, a strong positioning sequence, the so-called 601 sequence, exhibits rare, abrupt transitions of five and ten base pair steps without rotation. Moreover, we evaluated the importance of hydrogen bond interactions on the sliding mode, finding that strong and weak bonds favor respectively the rotation-coupled and -uncoupled sliding movements.

  10. Nucleosome loss leads to global transcriptional up-regulation and genomic instability during yeast aging.

    Science.gov (United States)

    Hu, Zheng; Chen, Kaifu; Xia, Zheng; Chavez, Myrriah; Pal, Sangita; Seol, Ja-Hwan; Chen, Chin-Chuan; Li, Wei; Tyler, Jessica K

    2014-02-15

    All eukaryotic cells divide a finite number of times, although the mechanistic basis of this replicative aging remains unclear. Replicative aging is accompanied by a reduction in histone protein levels, and this is a cause of aging in budding yeast. Here we show that nucleosome occupancy decreased by 50% across the whole genome during replicative aging using spike-in controlled micrococcal nuclease digestion followed by sequencing. Furthermore, nucleosomes became less well positioned or moved to sequences predicted to better accommodate histone octamers. The loss of histones during aging led to transcriptional induction of all yeast genes. Genes that are normally repressed by promoter nucleosomes were most induced, accompanied by preferential nucleosome loss from their promoters. We also found elevated levels of DNA strand breaks, mitochondrial DNA transfer to the nuclear genome, large-scale chromosomal alterations, translocations, and retrotransposition during aging.

  11. Generation of Native Chromatin Immunoprecipitation Sequencing Libraries for Nucleosome Density Analysis.

    Science.gov (United States)

    Lorzadeh, Alireza; Lopez Gutierrez, Rodrigo; Jackson, Linda; Moksa, Michelle; Hirst, Martin

    2017-12-12

    We present a modified native chromatin immunoprecipitation sequencing (ChIP-seq) experimental protocol compatible with a Gaussian mixture distribution based analysis methodology (nucleosome density ChIP-seq; ndChIP-seq) that enables the generation of combined measurements of micrococcal nuclease (MNase) accessibility with histone modification genome-wide. Nucleosome position and local density, and the posttranslational modification of their histone subunits, act in concert to regulate local transcription states. Combinatorial measurements of nucleosome accessibility with histone modification generated by ndChIP-seq allows for the simultaneous interrogation of these features. The ndChIP-seq methodology is applicable to small numbers of primary cells inaccessible to cross-linking based ChIP-seq protocols. Taken together, ndChIP-seq enables the measurement of histone modification in combination with local nucleosome density to obtain new insights into shared mechanisms that regulate RNA transcription within rare primary cell populations.

  12. Using DNA mechanics to predict in vitro nucleosome positions and formation energies

    National Research Council Canada - National Science Library

    Morozov, Alexandre V; Fortney, Karissa; Gaykalova, Daria A; Studitsky, Vasily M; Widom, Jonathan; Siggia, Eric D

    2009-01-01

    ... exclusion and sequence alone. We have developed a biophysical model, DNABEND, for the sequence dependence of DNA bending energies, and validated it against a collection of in vitro free energies of nucleosome formation and a set...

  13. Genome-wide nucleosome occupancy and DNA methylation profiling of four human cell lines

    Directory of Open Access Journals (Sweden)

    Aaron L. Statham

    2015-03-01

    Full Text Available DNA methylation and nucleosome positioning are two key mechanisms that contribute to the epigenetic control of gene expression. During carcinogenesis, the expression of many genes is altered alongside extensive changes in the epigenome, with repressed genes often being associated with local DNA hypermethylation and gain of nucleosomes at their promoters. However the spectrum of alterations that occur at distal regulatory regions has not been extensively studied. To address this we used Nucleosome Occupancy and Methylation sequencing (NOMe-seq to compare the genome-wide DNA methylation and nucleosome occupancy profiles between normal and cancer cell line models of the breast and prostate. Here we describe the bioinformatic pipeline and methods that we developed for the processing and analysis of the NOMe-seq data published by (Taberlay et al., 2014 [1] and deposited in the Gene Expression Omnibus with accession GSE57498.

  14. Understanding the connection between epigenetic DNA methylation and nucleosome positioning from computer simulations.

    Directory of Open Access Journals (Sweden)

    Guillem Portella

    Full Text Available Cytosine methylation is one of the most important epigenetic marks that regulate the process of gene expression. Here, we have examined the effect of epigenetic DNA methylation on nucleosomal stability using molecular dynamics simulations and elastic deformation models. We found that methylation of CpG steps destabilizes nucleosomes, especially when these are placed in sites where the DNA minor groove faces the histone core. The larger stiffness of methylated CpG steps is a crucial factor behind the decrease in nucleosome stability. Methylation changes the positioning and phasing of the nucleosomal DNA, altering the accessibility of DNA to regulatory proteins, and accordingly gene functionality. Our theoretical calculations highlight a simple physical-based explanation on the foundations of epigenetic signaling.

  15. Soft skills turned into hard facts: nucleosome remodelling at developmental switches.

    Science.gov (United States)

    Chioda, M; Becker, P B

    2010-07-01

    Nucleosome remodelling factors are regulators of DNA accessibility in chromatin and lubricators of all major functions of eukaryotic genomes. Their action is transient and reversible, yet can be decisive for irreversible cell-fate decisions during development. In addition to the well-known local actions of nucleosome remodelling factors during transcription initiation, more global and fundamental roles for remodelling complexes in shaping the epigenome during development are emerging.

  16. Serum nucleosomes during neoadjuvant chemotherapy in patients with cervical cancer. Predictive and prognostic significance

    Directory of Open Access Journals (Sweden)

    Cetina Lucely

    2005-06-01

    Full Text Available Abstract Background It has been shown that free DNA circulates in serum plasma of patients with cancer and that at least part is present in the form of oligo- and monucleosomes, a marker of cell death. Preliminary data has shown a good correlation between decrease of nucleosomes with response and prognosis. Here, we performed pre- and post-chemotherapy determinations of serum nucleosomes with an enzyme-linked immunosorbent assay (ELISA method in a group of patients with cervical cancer receiving neoadjuvant chemotherapy. Methods From December 2000 to June 2001, 41 patients with cervical cancer staged as FIGO stages IB2-IIIB received three 21-day courses of carboplatin and paclitaxel, both administered at day 1; then, patients underwent radical hysterectomy. Nucleosomes were measured the day before (baseline, at day seven of the first course and day seven of the third course of chemotherapy. Values of nucleosomes were analyzed with regard to pathologic response and to time to progression-free and overall survival. Results All patients completed chemotherapy, were evaluable for pathologic response, and had nucleosome levels determined. At a mean follow-up of 23 months (range, 7–26 months, projected progression time and overall survival were 80.3 and 80.4%, respectively. Mean differential values of nucleosomes were lower in the third course as compared with the first course (p >0.001. The decrease in the third course correlated with pathologic response (p = 0.041. Survival analysis showed a statistically significant, better progression-free and survival time in patients who showed lower levels at the third course (p = 0.0243 and p = 0.0260, respectively. Cox regression analysis demonstrated that nucleosome increase in the third course increased risk of death to 6.86 (95% confidence interval [CI 95%], 0.84–56.0. Conclusion Serum nucleosomes may have a predictive role for response and prognostic significance in patients with cervical cancer

  17. Anti-nucleosome antibodies as a disease activity marker in patients with systemic lupus erythematosus.

    Science.gov (United States)

    Suleiman, Suleiman; Kamaliah, Daud; Nadeem, Afzal; Naing, Nyi Nyi; Che Maraina, Che Hussin

    2009-07-01

    To measure the level of anti-nucleosome antibodies in systemic lupus erythematosus (SLE) patients, to determine the sensitivity and the specificity of these antibodies in the diagnosis of the disease and to evaluate the relationship between the levels of anti-nucleosome antibodies, anti-dsDNA (double-stranded DNA) and SLE disease activity. A cross-sectional study was conducted. All patients attended either a medical specialist clinic or were admitted to the medical wards of Hospital Universiti Sains Malaysia with the diagnosis of SLE (n = 90), other connective tissue diseases (n = 45) or were normal controls (n = 90) within the period from July 2004 until September 2005. They were tested for anti-nucleosome antibodies by enzyme-linked immunosorbent assay and anti-DNA antibodies by immunofluorescence. SLE disease activity was evaluated by SLE disease activity index (SLEDAI) score. Out of 90 SLE patients, anti-nucleosome antibodies were positive in 47 (52.2%) patients, whereas these antibodies were positive in three (6.7%) patients with other connective tissue diseases. Anti-dsDNA antibodies were positive in 33 (36.7%) SLE patients, whereas these antibodies were positive in four (8.9%) patients with other connective tissue diseases. Anti-nucleosome antibodies were positive in 40 (97.6%) patients with active SLE, whereas these antibodies were positive in seven (14.3%) patients with inactive SLE. Anti-nucleosome antibodies had a stronger correlation than anti-dsDNA antibodies with SLEDAI score. There was a significant association between anti-nucleosome antibodies and disease activity. Anti-nucleosome antibodies test is highly sensitive and specific for the diagnosis of SLE, especially when the anti-dsDNA antibodies are absent. They are additional disease activity markers in the assessment of SLE disease activity.

  18. A quantitative investigation of linker histone interactions with nucleosomes and chromatin

    Science.gov (United States)

    White, Alison E.; Hieb, Aaron R.; Luger, Karolin

    2016-01-01

    Linker histones such as H1 are abundant basic proteins that bind tightly to nucleosomes, thereby acting as key organizers of chromatin structure. The molecular details of linker histone interactions with the nucleosome, and in particular the contributions of linker DNA and of the basic C-terminal tail of H1, are controversial. Here we combine rigorous solution-state binding assays with native gel electrophoresis and Atomic Force Microscopy, to quantify the interaction of H1 with chromatin. We find that H1 binds nucleosomes and nucleosomal arrays with very tight affinity by recognizing a specific DNA geometry minimally consisting of a solitary nucleosome with a single ~18 base pair DNA linker arm. The association of H1 alters the conformation of trinucleosomes so that only one H1 can bind to the two available linker DNA regions. Neither incorporation of the histone variant H2A.Z, nor the presence of neighboring nucleosomes affects H1 affinity. Our data provide a comprehensive thermodynamic framework for this ubiquitous chromatin architectural protein. PMID:26750377

  19. Inducible nucleosome depletion at OREBP-binding-sites by hypertonic stress.

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    Edith H Y Tong

    Full Text Available BACKGROUND: Osmotic Response Element-Binding Protein (OREBP, also known as TonEBP or NFAT5, is a unique transcription factor. It is hitherto the only known mammalian transcription factor that regulates hypertonic stress-induced gene transcription. In addition, unlike other monomeric members of the NFAT family, OREBP exists as a homodimer and it is the only transcription factor known to bind naked DNA targets by complete encirclement in vitro. Nevertheless, how OREBP interacts with target DNA, also known as ORE/TonE, and how it elicits gene transcription in vivo, remains unknown. METHODOLOGY: Using hypertonic induction of the aldose reductase (AR gene activation as a model, we showed that OREs contained dynamic nucleosomes. Hypertonic stress induced a rapid and reversible loss of nucleosome(s around the OREs. The loss of nucleosome(s was found to be initiated by an OREBP-independent mechanism, but was significantly potentiated in the presence of OREBP. Furthermore, hypertonic induction of AR gene was associated with an OREBP-dependent hyperacetylation of histones that spanned the 5' upstream sequences and at least some exons of the gene. Nevertheless, nucleosome loss was not regulated by the acetylation status of histone. SIGNIFICANCE: Our findings offer novel insights into the mechanism of OREBP-dependent transcriptional regulation and provide a basis for understanding how histone eviction and transcription factor recruitment are coupled.

  20. Charge State of the Globular Histone Core Controls Stability of the Nucleosome

    Science.gov (United States)

    Fenley, Andrew T.; Adams, David A.; Onufriev, Alexey V.

    2010-01-01

    Presented here is a quantitative model of the wrapping and unwrapping of the DNA around the histone core of the nucleosome that suggests a mechanism by which this transition can be controlled: alteration of the charge state of the globular histone core. The mechanism is relevant to several classes of posttranslational modifications such as histone acetylation and phosphorylation; several specific scenarios consistent with recent in vivo experiments are considered. The model integrates a description based on an idealized geometry with one based on the atomistic structure of the nucleosome, and the model consistently accounts for both the electrostatic and nonelectrostatic contributions to the nucleosome free energy. Under physiological conditions, isolated nucleosomes are predicted to be very stable (38 ± 7 kcal/mol). However, a decrease in the charge of the globular histone core by one unit charge, for example due to acetylation of a single lysine residue, can lead to a significant decrease in the strength of association with its DNA. In contrast to the globular histone core, comparable changes in the charge state of the histone tail regions have relatively little effect on the nucleosome's stability. The combination of high stability and sensitivity explains how the nucleosome is able to satisfy the seemingly contradictory requirements for thermodynamic stability while allowing quick access to its DNA informational content when needed by specific cellular processes such as transcription. PMID:20816070

  1. Nucleosome Repositioning: A Novel Mechanism for Nicotine- and Cocaine-Induced Epigenetic Changes.

    Directory of Open Access Journals (Sweden)

    Amber N Brown

    Full Text Available Drugs of abuse modify behavior by altering gene expression in the brain. Gene expression can be regulated by changes in DNA methylation as well as by histone modifications, which alter chromatin structure, DNA compaction and DNA accessibility. In order to better understand the molecular mechanisms directing drug-induced changes in chromatin structure, we examined DNA-nucleosome interactions within promoter regions of 858 genes in human neuroblastoma cells (SH-SY5Y exposed to nicotine or cocaine. Widespread, drug- and time-resolved repositioning of nucleosomes was identified at the transcription start site and promoter region of multiple genes. Nicotine and cocaine produced unique and shared changes in terms of the numbers and types of genes affected, as well as repositioning of nucleosomes at sites which could increase or decrease the probability of gene expression based on DNA accessibility. Half of the drug-induced nucleosome positions approximated a theoretical model of nucleosome occupancy based on physical and chemical characteristics of the DNA sequence, whereas the basal or drug naïve positions were generally DNA sequence independent. Thus we suggest that nucleosome repositioning represents an initial dynamic genome-wide alteration of the transcriptional landscape preceding more selective downstream transcriptional reprogramming, which ultimately characterizes the cell- and tissue-specific responses to drugs of abuse.

  2. Nucleosome occupancy as a novel chromatin parameter for replication origin functions

    Science.gov (United States)

    Rodriguez, Jairo; Lee, Laura; Lynch, Bryony; Tsukiyama, Toshio

    2017-01-01

    Eukaryotic DNA replication initiates from multiple discrete sites in the genome, termed origins of replication (origins). Prior to S phase, multiple origins are poised to initiate replication by recruitment of the pre-replicative complex (pre-RC). For proper replication to occur, origin activation must be tightly regulated. At the population level, each origin has a distinct firing time and frequency of activation within S phase. Many studies have shown that chromatin can strongly influence initiation of DNA replication. However, the chromatin parameters that affect properties of origins have not been thoroughly established. We found that nucleosome occupancy in G1 varies greatly around origins across the S. cerevisiae genome, and nucleosome occupancy around origins significantly correlates with the activation time and efficiency of origins, as well as pre-RC formation. We further demonstrate that nucleosome occupancy around origins in G1 is established during transition from G2/M to G1 in a pre-RC-dependent manner. Importantly, the diminished cell-cycle changes in nucleosome occupancy around origins in the orc1-161 mutant are associated with an abnormal global origin usage profile, suggesting that proper establishment of nucleosome occupancy around origins is a critical step for regulation of global origin activities. Our work thus establishes nucleosome occupancy as a novel and key chromatin parameter for proper origin regulation. PMID:27895110

  3. GAA triplet-repeats cause nucleosome depletion in the human genome.

    Science.gov (United States)

    Zhao, Hongyu; Xing, Yongqiang; Liu, Guoqing; Chen, Ping; Zhao, Xiujuan; Li, Guohong; Cai, Lu

    2015-08-01

    Although there have been many investigations into how trinucleotide repeats affect nucleosome formation and local chromatin structure, the nucleosome positioning of GAA triplet-repeats in the human genome has remained elusive. In this work, the nucleosome occupancy around GAA triplet-repeats across the human genome was computed statistically. The results showed a nucleosome-depleted region in the vicinity of GAA triplet-repeats in activated and resting CD4(+) T cells. Furthermore, the A-tract was frequently adjacent to the upstream region of GAA triplet-repeats and could enhance the depletion surrounding GAA triplet-repeats. In vitro chromatin reconstitution assays with GAA-containing plasmids also demonstrated that the inserted GAA triplet-repeats destabilized the ability of recombinant plasmids to assemble nucleosomes. Our results suggested that GAA triplet-repeats have lower affinity to histones and can change local nucleosome positioning. These findings may be helpful for understanding the mechanism of Friedreich's ataxia, which is associated with GAA triplet-repeats at the chromatin level. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Crystal structure of the nucleosome containing ultraviolet light-induced cyclobutane pyrimidine dimer.

    Science.gov (United States)

    Horikoshi, Naoki; Tachiwana, Hiroaki; Kagawa, Wataru; Osakabe, Akihisa; Matsumoto, Syota; Iwai, Shigenori; Sugasawa, Kaoru; Kurumizaka, Hitoshi

    2016-02-26

    The cyclobutane pyrimidine dimer (CPD) is induced in genomic DNA by ultraviolet (UV) light. In mammals, this photolesion is primarily induced within nucleosomal DNA, and repaired exclusively by the nucleotide excision repair (NER) pathway. However, the mechanism by which the CPD is accommodated within the nucleosome has remained unknown. We now report the crystal structure of a nucleosome containing CPDs. In the nucleosome, the CPD induces only limited local backbone distortion, and the affected bases are accommodated within the duplex. Interestingly, one of the affected thymine bases is located within 3.0 Å from the undamaged complementary adenine base, suggesting the formation of complementary hydrogen bonds in the nucleosome. We also found that UV-DDB, which binds the CPD at the initial stage of the NER pathway, also efficiently binds to the nucleosomal CPD. These results provide important structural and biochemical information for understanding how the CPD is accommodated and recognized in chromatin. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. A bromodomain–DNA interaction facilitates acetylation-dependent bivalent nucleosome recognition by the BET protein BRDT

    Science.gov (United States)

    Miller, Thomas C. R.; Simon, Bernd; Rybin, Vladimir; Grötsch, Helga; Curtet, Sandrine; Khochbin, Saadi; Carlomagno, Teresa; Müller, Christoph W.

    2016-01-01

    Bromodomains are critical components of many chromatin modifying/remodelling proteins and are emerging therapeutic targets, yet how they interact with nucleosomes, rather than acetylated peptides, remains unclear. Using BRDT as a model, we characterized how the BET family of bromodomains interacts with site-specifically acetylated nucleosomes. Here we report that BRDT interacts with nucleosomes through its first (BD1), but not second (BD2) bromodomain, and that acetylated histone recognition by BD1 is complemented by a bromodomain–DNA interaction. Simultaneous DNA and histone recognition enhances BRDT's nucleosome binding affinity and specificity, and its ability to localize to acetylated chromatin in cells. Conservation of DNA binding in bromodomains of BRD2, BRD3 and BRD4, indicates that bivalent nucleosome recognition is a key feature of these bromodomains and possibly others. Our results elucidate the molecular mechanism of BRDT association with nucleosomes and identify structural features of the BET bromodomains that may be targeted for therapeutic inhibition. PMID:27991587

  6. Increased Nucleosomes and Neutrophil Activation Link to Disease Progression in Patients with Scrub Typhus but Not Murine Typhus in Laos.

    Directory of Open Access Journals (Sweden)

    Daniel H Paris

    Full Text Available Cell-mediated immunity is essential in protection against rickettsial illnesses, but the role of neutrophils in these intracellular vasculotropic infections remains unclear. This study analyzed the plasma levels of nucleosomes, FSAP-activation (nucleosome-releasing factor, and neutrophil activation, as evidenced by neutrophil-elastase (ELA complexes, in sympatric Lao patients with scrub typhus and murine typhus. In acute scrub typhus elevated nucleosome levels correlated with lower GCS scores, raised respiratory rate, jaundice and impaired liver function, whereas neutrophil activation correlated with fibrinolysis and high IL-8 plasma levels, a recently identified predictor of severe disease and mortality. Nucleosome and ELA complex levels were associated with a 4.8-fold and 4-fold increased risk of developing severe scrub typhus, beyond cut off values of 1,040 U/ml for nucleosomes and 275 U/ml for ELA complexes respectively. In murine typhus, nucleosome levels associated with pro-inflammatory cytokines and the duration of illness, while ELA complexes correlated strongly with inflammation markers, jaundice and increased respiratory rates. This study found strong correlations between circulating nucleosomes and neutrophil activation in patients with scrub typhus, but not murine typhus, providing indirect evidence that nucleosomes could originate from neutrophil extracellular trap (NET degradation. High circulating plasma nucleosomes and ELA complexes represent independent risk factors for developing severe complications in scrub typhus. As nucleosomes and histones exposed on NETs are highly cytotoxic to endothelial cells and are strongly pro-coagulant, neutrophil-derived nucleosomes could contribute to vascular damage, the pro-coagulant state and exacerbation of disease in scrub typhus, thus indicating a detrimental role of neutrophil activation. The data suggest that increased neutrophil activation relates to disease progression and severe

  7. PuFFIN--a parameter-free method to build nucleosome maps from paired-end reads.

    Science.gov (United States)

    Polishko, Anton; Bunnik, Evelien M; Le Roch, Karine G; Lonardi, Stefano

    2014-01-01

    We introduce a novel method, called PuFFIN, that takes advantage of paired-end short reads to build genome-wide nucleosome maps with larger numbers of detected nucleosomes and higher accuracy than existing tools. In contrast to other approaches that require users to optimize several parameters according to their data (e.g., the maximum allowed nucleosome overlap or legal ranges for the fragment sizes) our algorithm can accurately determine a genome-wide set of non-overlapping nucleosomes without any user-defined parameter. This feature makes PuFFIN significantly easier to use and prevents users from choosing the "wrong" parameters and obtain sub-optimal nucleosome maps. PuFFIN builds genome-wide nucleosome maps using a multi-scale (or multi-resolution) approach. Our algorithm relies on a set of nucleosome "landscape" functions at different resolution levels: each function represents the likelihood of each genomic location to be occupied by a nucleosome for a particular value of the smoothing parameter. After a set of candidate nucleosomes is computed for each function, PuFFIN produces a consensus set that satisfies non-overlapping constraints and maximizes the number of nucleosomes. We report comprehensive experimental results that compares PuFFIN with recently published tools (NOrMAL, TEMPLATE FILTERING, and NucPosSimulator) on several synthetic datasets as well as real data for S. cerevisiae and P. falciparum. Experimental results show that our approach produces more accurate nucleosome maps with a higher number of non-overlapping nucleosomes than other tools.

  8. Increasing Nucleosome Occupancy Is Correlated with an Increasing Mutation Rate so Long as DNA Repair Machinery Is Intact.

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    Puya G Yazdi

    Full Text Available Deciphering the multitude of epigenomic and genomic factors that influence the mutation rate is an area of great interest in modern biology. Recently, chromatin has been shown to play a part in this process. To elucidate this relationship further, we integrated our own ultra-deep sequenced human nucleosomal DNA data set with a host of published human genomic and cancer genomic data sets. Our results revealed, that differences in nucleosome occupancy are associated with changes in base-specific mutation rates. Increasing nucleosome occupancy is associated with an increasing transition to transversion ratio and an increased germline mutation rate within the human genome. Additionally, cancer single nucleotide variants and microindels are enriched within nucleosomes and both the coding and non-coding cancer mutation rate increases with increasing nucleosome occupancy. There is an enrichment of cancer indels at the theoretical start (74 bp and end (115 bp of linker DNA between two nucleosomes. We then hypothesized that increasing nucleosome occupancy decreases access to DNA by DNA repair machinery and could account for the increasing mutation rate. Such a relationship should not exist in DNA repair knockouts, and we thus repeated our analysis in DNA repair machinery knockouts to test our hypothesis. Indeed, our results revealed no correlation between increasing nucleosome occupancy and increasing mutation rate in DNA repair knockouts. Our findings emphasize the linkage of the genome and epigenome through the nucleosome whose properties can affect genome evolution and genetic aberrations such as cancer.

  9. Friedreich's ataxia GAA.TTC duplex and GAA.GAA.TTC triplex structures exclude nucleosome assembly.

    Science.gov (United States)

    Ruan, Haihe; Wang, Yuh-Hwa

    2008-11-07

    Both chromatin structure and formation of triplex DNA at expanded GAA TTC repeats have been shown to regulate the FXN gene silencing, which causes Friedreich's ataxia. Recent studies have suggested that the presence of heterochromatin at the long expanded GAA TTC repeats, which is enriched in hypoacetylated histones, deters the transcription of the FXN gene. However, neither direct histone binding nor the effect of histone acetylation on the GAA TTC duplex or the GAA GAA TTC triplex has been measured in vitro. In this study, GAA TTC repeating DNAs derived from the human FXN gene, and the GAA GAA TTC triplex, were examined for their ability to assemble single nucleosomes and nucleosome arrays. Competitive nucleosome reconstitution assays demonstrated that the GAA TTC duplex excludes nucleosomes (53% decrease compared to the pUC control DNA) and that the GAA GAA TTC triplex further lowers the nucleosome assembly efficiency (82% decrease compared to the duplex DNA). The difference in assembly efficiency is amplified more significantly when hypoacetylated histones are used, compared to assembly with hyperacetylated histones. By analyzing the formation of nucleosome arrays on GAA TTC-containing plasmids, the triplex structure was shown to destabilize the ability of adjacent sequences to assemble nucleosomes. These results provide the first direct binding measurements for the GAA TTC duplex and the GAA GAA TTC triplex, and on the effect of histone acetylation, towards dissecting the role of chromatin structure in silencing of the FXN gene. These findings suggest that these sequences could profoundly alter local chromatin structure, and the discrepancy between in vivo and in vitro results supports recent studies showing that, in addition to DNA sequences, other factors such as epigenetic marks could be involved in the mechanism for inhibition of FXN gene expression.

  10. Transport of nucleosome core particles in semidilute DNA solutions.

    Science.gov (United States)

    Mangenot, Stéphanie; Keller, Simon; Rädler, Joachim

    2003-09-01

    We studied the diffusion of native and trypsinized nucleosome core particles (NCPs), in aqueous solution and in concentrated DNA solutions (0.25-100 mg/ml) using fluorescence correlation spectroscopy (FCS). The highest DNA concentrations studied mimic the DNA density inside the cell nucleus. The diffusion coefficient of freely diffusing NCPs depends on the presence or absence of histone tails and is affected by the salt concentration due to the relaxation effect of counterions. NCPs placed in a network of long DNA molecules (30-50 kbp) reveal anomalous diffusion. We demonstrate that NCPs diffusion is in agreement with known particle transport in entangled macromolecular solutions as long as the histone tails are folded onto the particles. In contrast, when these tails are unfolded, the reversible adsorption of NCPs onto the DNA network has to be taken into account. This is confirmed by the fact that removal of the tails leads to reduction of the interaction between NCPs and the DNA network. The findings suggest that histone tail bridging plays an important role in chromatin dynamics.

  11. Structure of centromere chromatin: from nucleosome to chromosomal architecture.

    Science.gov (United States)

    Schalch, Thomas; Steiner, Florian A

    2017-08-01

    The centromere is essential for the segregation of chromosomes, as it serves as attachment site for microtubules to mediate chromosome segregation during mitosis and meiosis. In most organisms, the centromere is restricted to one chromosomal region that appears as primary constriction on the condensed chromosome and is partitioned into two chromatin domains: The centromere core is characterized by the centromere-specific histone H3 variant CENP-A (also called cenH3) and is required for specifying the centromere and for building the kinetochore complex during mitosis. This core region is generally flanked by pericentric heterochromatin, characterized by nucleosomes containing H3 methylated on lysine 9 (H3K9me) that are bound by heterochromatin proteins. During mitosis, these two domains together form a three-dimensional structure that exposes CENP-A-containing chromatin to the surface for interaction with the kinetochore and microtubules. At the same time, this structure supports the tension generated during the segregation of sister chromatids to opposite poles. In this review, we discuss recent insight into the characteristics of the centromere, from the specialized chromatin structures at the centromere core and the pericentromere to the three-dimensional organization of these regions that make up the functional centromere.

  12. Molecular Shapes From Rotational Diffusion: Dye Molecules, Proteins And Nucleosomes

    Science.gov (United States)

    Small, Enoch W.; Libertini, Louis J.; Small, Jeanne R.

    1988-06-01

    This paper examines the shape information available from measurements of fluorescence anisotropy decays. We report anisotropy decays for three systems and examine the kinds of information which may be obtained. Results for the dye rose bengal (~1000 dal-tons) suggest that an approximation used in rotational diffusion theory, that the solvent molecules are very small compared to the solute, is valid even for this relatively small molecule. Second, we examine the effects of calcium concentration on rotational diffusion of the protein calmodulin (~17,000 daltons) derivatized by crosslinking to form a dityrosine fluorophore. In the presence of sufficiently high calcium ion concentration, this crosslinked calmodulin shows a single exponential anisotropy decay which indicates rotational diffusion consistent with the extended dumbbell structure found for calmodulin by x-ray crystal-lography. At low calcium concentrations, th e crosslinked calmodulin rotates considerably faster, suggesting a much more compact shape; also, this anisotropy decay includes considerably shorter correlation times which are interpreted as arising from a segmental flexibility not evident for crosslinked calmodulin at high calcium concentration. Finally, we examine a transition which is observed at very low salt concentrations for nucleosome core particles, relatively large complexes (~200,000 daltons) derived from chromatin and comprised of eight histone molecules and 145 base pairs of DNA. The anisotropy decay of ethidium bound to the DNA indicates that core particles exposed to low ionic strength are considerably elongated relative to the shape at higher ionic strength.

  13. Histone h3 glutathionylation in proliferating mammalian cells destabilizes nucleosomal structure.

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    García-Giménez, José Luis; Òlaso, Gloria; Hake, Sandra B; Bönisch, Clemens; Wiedemann, Sonja M; Markovic, Jelena; Dasí, Francisco; Gimeno, Amparo; Pérez-Quilis, Carme; Palacios, Oscar; Capdevila, Mercè; Viña, José; Pallardó, Federico V

    2013-10-20

    Here we report that chromatin, the complex and dynamic eukaryotic DNA packaging structure, is able to sense cellular redox changes. Histone H3, the only nucleosomal protein that possesses cysteine(s), can be modified by glutathione (GSH). Using Biotin labeled glutathione ethyl ester (BioGEE) treatment of nucleosomes in vitro, we show that GSH, the most abundant antioxidant in mammals, binds to histone H3. BioGEE treatment of NIH3T3 cells indicates that glutathionylation of H3 is maximal in fast proliferating cells, correlating well with enhanced levels of H3 glutathionylation in different tumor cell lines. Furthermore, glutathionylation of H3 in vivo decreases in livers from aged SAMP8 and C57BL/6J mice. We demonstrate biochemically and by mass spectrometry that histone variants H3.2/H3.3 are glutathionylated on their cysteine residue 110. Furthermore, circular dichroism, thermal denaturation of reconstituted nucleosomes, and molecular modeling indicate that glutathionylation of histone H3 produces structural changes affecting nucleosomal stability. We characterize the implications of histone H3 glutathionylation in cell physiology and the modulation of core histone proteins structure affected by this modification. Histone H3 senses cellular redox changes through glutathionylation of Cys, which increases during cell proliferation and decreases during aging. Glutathionylation of histone H3 affects nucleosome stability structure leading to a more open chromatin structure.

  14. Crystal structure of the PRC1 ubiquitylation module bound to the nucleosome.

    Science.gov (United States)

    McGinty, Robert K; Henrici, Ryan C; Tan, Song

    2014-10-30

    The Polycomb group of epigenetic enzymes represses expression of developmentally regulated genes in many eukaryotes. This group includes the Polycomb repressive complex 1 (PRC1), which ubiquitylates nucleosomal histone H2A Lys 119 using its E3 ubiquitin ligase subunits, Ring1B and Bmi1, together with an E2 ubiquitin-conjugating enzyme, UbcH5c. However, the molecular mechanism of nucleosome substrate recognition by PRC1 or other chromatin enzymes is unclear. Here we present the crystal structure of the human Ring1B-Bmi1-UbcH5c E3-E2 complex (the PRC1 ubiquitylation module) bound to its nucleosome core particle substrate. The structure shows how a chromatin enzyme achieves substrate specificity by interacting with several nucleosome surfaces spatially distinct from the site of catalysis. Our structure further reveals an unexpected role for the ubiquitin E2 enzyme in substrate recognition, and provides insight into how the related histone H2A E3 ligase, BRCA1, interacts with and ubiquitylates the nucleosome.

  15. Nucleosome assembly protein-1 is a linker histone chaperone in Xenopus eggs.

    Science.gov (United States)

    Shintomi, Keishi; Iwabuchi, Mari; Saeki, Hideaki; Ura, Kiyoe; Kishimoto, Takeo; Ohsumi, Keita

    2005-06-07

    In eukaryotic cells, genomic DNA is primarily packaged into nucleosomes through sequential ordered binding of the core and linker histone proteins. The acidic proteins termed histone chaperones are known to bind to core histones to neutralize their positive charges, thereby facilitating their proper deposition onto DNA to assemble the core of nucleosomes. For linker histones, however, little has been known about the regulatory mechanism for deposition of linker histones onto the linker DNA. Here we report that, in Xenopus eggs, the linker histone is associated with the Xenopus homologue of nucleosome assembly protein-1 (NAP-1), which is known to be a chaperone for the core histones H2A and H2B in Drosophila and mammalian cells [Ito, T., Bulger, M., Kobayashi, R. & Kadonaga, J. T. (1996) Mol. Cell Biol. 16, 3112-3124; Chang, L., Loranger, S. S., Mizzen, C., Ernst, S. G., Allis, C. D. & Annunziato, A. T. (1997) Biochemistry 36, 469-480]. We show that NAP-1 acts as the chaperone for the linker histone in both sperm chromatin remodeling into nucleosomes and linker histone binding to nucleosome core dimers. In the presence of NAP-1, the linker histone is properly deposited onto linker DNA at physiological ionic strength, without formation of nonspecific aggregates. These results strongly suggest that NAP-1 functions as a chaperone for the linker histone in Xenopus eggs.

  16. Nucleosome Assembly Dynamics Involve Spontaneous Fluctuations in the Handedness of Tetrasomes

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    Rifka Vlijm

    2015-01-01

    Full Text Available DNA wrapping around histone octamers generates nucleosomes, the basic compaction unit of eukaryotic chromatin. Nucleosome stability is carefully tuned to maintain DNA accessibility in transcription, replication, and repair. Using freely orbiting magnetic tweezers, which measure the twist and length of single DNA molecules, we monitor the real-time loading of tetramers or complete histone octamers onto DNA by Nucleosome Assembly Protein-1 (NAP1. Remarkably, we find that tetrasomes exhibit spontaneous flipping between a preferentially occupied left-handed state (ΔLk = −0.73 and a right-handed state (ΔLk = +1.0, separated by a free energy difference of 2.3 kBT (1.5 kcal/mol. This flipping occurs without concomitant changes in DNA end-to-end length. The application of weak positive torque converts left-handed tetrasomes into right-handed tetrasomes, whereas nucleosomes display more gradual conformational changes. Our findings reveal unexpected dynamical rearrangements of the nucleosomal structure, suggesting that chromatin can serve as a “twist reservoir,” offering a mechanistic explanation for the regulation of DNA supercoiling in chromatin.

  17. Distinct Roles of Histone H3 and H2A Tails in Nucleosome Stability

    Science.gov (United States)

    Li, Zhenhai; Kono, Hidetoshi

    2016-01-01

    Nucleosome breathing potentially increases the DNA exposure, which in turn recruits DNA-binding protein and regulates gene transcription. Numerous studies have shown the critical roles of N-terminal tails of histones H3 and H4 in gene expression; however, few studies have focused on the H2A C-terminal tail. Here we present thorough computational studies on a single nucleosome particle showing the linker DNA closing and opening, which is thought to be nucleosome breathing. With our simulation, the H2A C-terminal and H3 N-terminal tails were found to modulate the nucleosome conformation differently. The H2A C-terminal tail regulates nucleosome conformation by binding to linker DNA at different locations, whereas the H3 N-terminal tail regulates linker DNA by binding to it in different patterns. Further MD simulation on tail truncated structures corroborates this analysis. These findings replenish our understanding of the histone tail regulation mechanism on atomic level. PMID:27527579

  18. The dynamics of individual nucleosomes controls the chromatin condensation pathway: direct atomic force microscopy visualization of variant chromatin.

    Science.gov (United States)

    Montel, Fabien; Menoni, Hervé; Castelnovo, Martin; Bednar, Jan; Dimitrov, Stefan; Angelov, Dimitar; Faivre-Moskalenko, Cendrine

    2009-07-22

    Chromatin organization and dynamics is studied at scales ranging from single nucleosome to nucleosomal array by using a unique combination of biochemical assays, single molecule imaging technique, and numerical modeling. We show that a subtle modification in the nucleosome structure induced by the histone variant H2A.Bbd drastically modifies the higher order organization of the nucleosomal arrays. Importantly, as directly visualized by atomic force microscopy, conventional H2A nucleosomal arrays exhibit specific local organization, in contrast to H2A.Bbd arrays, which show "beads on a string" structure. The combination of systematic image analysis and theoretical modeling allows a quantitative description relating the observed gross structural changes of the arrays to their local organization. Our results suggest strongly that higher-order organization of H1-free nucleosomal arrays is determined mainly by the fluctuation properties of individual nucleosomes. Moreover, numerical simulations suggest the existence of attractive interactions between nucleosomes to provide the degree of compaction observed for conventional chromatin fibers.

  19. Time between onset of apoptosis and release of nucleosomes from apoptotic cells: putative implications for systemic lupus erythematosus

    NARCIS (Netherlands)

    van Nieuwenhuijze, A. E. M.; van Lopik, T.; Smeenk, R. J. T.; Aarden, L. A.

    2003-01-01

    OBJECTIVE: To investigate the kinetics of nucleosome leakage from apoptotic cells in an in vitro system and extrapolate the results to autoimmune disease, in particular systemic lupus erythematosus. METHODS: A sensitive nucleosome enzyme linked immunosorbent assay (ELISA) was developed, using a

  20. Physical properties of naked DNA influence nucleosome positioning and correlate with transcription start and termination sites in yeast

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    Soler-López Montserrat

    2011-10-01

    Full Text Available Abstract Background In eukaryotic organisms, DNA is packaged into chromatin structure, where most of DNA is wrapped into nucleosomes. DNA compaction and nucleosome positioning have clear functional implications, since they modulate the accessibility of genomic regions to regulatory proteins. Despite the intensive research effort focused in this area, the rules defining nucleosome positioning and the location of DNA regulatory regions still remain elusive. Results Naked (histone-free and nucleosomal DNA from yeast were digested by microccocal nuclease (MNase and sequenced genome-wide. MNase cutting preferences were determined for both naked and nucleosomal DNAs. Integration of their sequencing profiles with DNA conformational descriptors derived from atomistic molecular dynamic simulations enabled us to extract the physical properties of DNA on a genomic scale and to correlate them with chromatin structure and gene regulation. The local structure of DNA around regulatory regions was found to be unusually flexible and to display a unique pattern of nucleosome positioning. Ab initio physical descriptors derived from molecular dynamics were used to develop a computational method that accurately predicts nucleosome enriched and depleted regions. Conclusions Our experimental and computational analyses jointly demonstrate a clear correlation between sequence-dependent physical properties of naked DNA and regulatory signals in the chromatin structure. These results demonstrate that nucleosome positioning around TSS (Transcription Start Site and TTS (Transcription Termination Site (at least in yeast is strongly dependent on DNA physical properties, which can define a basal regulatory mechanism of gene expression.

  1. Nucleosome Binding Alters the Substrate Bonding Environment of Histone H3 Lysine 36 Methyltransferase NSD2.

    Science.gov (United States)

    Poulin, Myles B; Schneck, Jessica L; Matico, Rosalie E; Hou, Wangfang; McDevitt, Patrick J; Holbert, Marc; Schramm, Vern L

    2016-06-01

    Nuclear receptor-binding SET domain protein 2 (NSD2) is a histone H3 lysine 36 (H3K36)-specific methyltransferase enzyme that is overexpressed in a number of cancers, including multiple myeloma. NSD2 binds to S-adenosyl-l-methionine (SAM) and nucleosome substrates to catalyze the transfer of a methyl group from SAM to the ε-amino group of histone H3K36. Equilibrium binding isotope effects and density functional theory calculations indicate that the SAM methyl group is sterically constrained in complex with NSD2, and that this steric constraint is released upon nucleosome binding. Together, these results show that nucleosome binding to NSD2 induces a significant change in the chemical environment of enzyme-bound SAM.

  2. Extra views on structure and dynamics of DNA loops on nucleosomes studied with molecular simulations.

    Science.gov (United States)

    Pasi, Marco; Angelov, Dimitar; Bednar, Jan; Dimitrov, Stefan; Lavery, Richard

    2016-11-01

    It has been shown experimentally that the action of the RSC chromatin remodeler leads to the formation of an irregular, partially remodeled nucleosome, termed a remosome. The remosome contains an extra 30-40 base pairs of DNA compared to a canonical nucleosome. Large-scale molecular simulations have provided information on the probable structure of remosomes and have explained why they remain stable in the absence of RSC. Here we explain how these simulations were carried out and what the resulting remosome models imply in terms of the mechanism of action of RSC. We notably show that local kinks within DNA are key in explaining how extra DNA can be in added to nucleosomes without unduly disturbing DNA-histone binding.

  3. a Multi-Layer Model to Study Genome-Scale Positions of Nucleosomes

    Science.gov (United States)

    di Gesú, Vito; Lo Bosco, Giosué; Pinello, Luca; Corona, Davide; Collesano, Marianna; Yuan, Guo-Cheng

    2007-12-01

    The positioning of nucleosomes along chromatin has been implicated in the regulation of gene expression in eukaryotic cells, because packaging DNA into nucleosomes affects sequence accessibility. In this paper we propose a new model (called MLM) for the identification of nucleosomes and linker regions across DNA, consisting in a thresholding technique based on cut-set conditions. For this purpose we have defined a method to generate synthetic microarray data fully inspired from the approach that has been used by Yuan et al. [3]. Results have shown a good recognition rate on synthetic data, moreover, the MLM shows a good agreement with the recently published method based on Hidden Markov Model when tested on the Saccharomyces cerevisiae chromosomes microarray data.

  4. Role of histone tails in the conformation and interactions of nucleosome core particles.

    Science.gov (United States)

    Bertin, Aurélie; Leforestier, Amélie; Durand, Dominique; Livolant, Françoise

    2004-04-27

    The goal of this work was to test the role of the histone tails in the emergence of attractive interactions between nucleosomes above a critical salt concentration that corresponds to the complete tail extension outside the nucleosome [Mangenot, S., et al (2002) Biophys. J. 82, 345-356; Mangenot, S., et al (2002) Eur. Phys. J. E 7, 221-231]. Small angle X-ray scattering experiments were performed in parallel with intact and trypsin tail-deleted nucleosomes with 146 +/- 3 bp DNA. We varied the monovalent salt concentration from 10 to 300 monovalent salt concentration and followed the evolution of (i) the second virial coefficient that characterizes the interactions between particles and (ii) the conformation of the particle. The attractive interactions do not emerge in the absence of the tails, which validates the proposed hypothesis.

  5. New insights into nucleosome and chromatin structure: an ordered state or a disordered affair?

    Science.gov (United States)

    Luger, Karolin; Dechassa, Mekonnen L.; Tremethick, David J.

    2012-01-01

    The compaction of genomic DNA into chromatin has profound implications for the regulation of key processes such as transcription, replication and DNA repair. Nucleosomes, the repeating building blocks of chromatin, vary in the composition of their histone protein components. This is the result of the incorporation of variant histones and post-translational modifications of histone amino acid side chains. The resulting changes in nucleosome structure, stability and dynamics affect the compaction of nucleosomal arrays into higher-order structures. It is becoming clear that chromatin structures are not nearly as uniform and regular as previously assumed. This implies that chromatin structure must also be viewed in the context of specific biological functions. PMID:22722606

  6. The mechanism of nucleosome traversal by RNA polymerase II: roles for template uncoiling and transcript elongation factors.

    Science.gov (United States)

    Luse, Donal S; Studitsky, Vasily M

    2011-01-01

    RNA polymerase II traverses nucleosomes rapidly and efficiently in the cell but it has not been possible to duplicate this process in the test tube. A single nucleosome has generally been found to provide a strong barrier to transcript elongation in vitro. Recent studies have shown that effective transcript elongation can occur on nucleosomal templates in vitro, but this depends on both facilitated uncoiling of DNA from the octamer surface and the presence of transcription factors that maintain polymerase in the transcriptionally competent state. These findings indicate that the efficiency and rate of transcription through chromatin could be regulated through controlled DNA uncoiling. These studies also demonstrate that nucleosome traversal need not result in nucleosome displacement.

  7. ATP-Dependent Chromatin Remodeling Factors and Their Roles in Affecting Nucleosome Fiber Composition

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    Alexandra Lusser

    2011-10-01

    Full Text Available ATP-dependent chromatin remodeling factors of the SNF2 family are key components of the cellular machineries that shape and regulate chromatin structure and function. Members of this group of proteins have broad and heterogeneous functions ranging from controlling gene activity, facilitating DNA damage repair, promoting homologous recombination to maintaining genomic stability. Several chromatin remodeling factors are critical components of nucleosome assembly processes, and recent reports have identified specific functions of distinct chromatin remodeling factors in the assembly of variant histones into chromatin. In this review we will discuss the specific roles of ATP-dependent chromatin remodeling factors in determining nucleosome composition and, thus, chromatin fiber properties.

  8. Recent insights from single-molecule studies into nucleosome structure and dynamics

    NARCIS (Netherlands)

    Ordu, O.; Lusser, A; Dekker, N.H.

    2016-01-01

    Eukaryotic DNA is tightly packed into a hierarchically ordered structure called chromatin in order to fit into the micron-scaled nucleus. The basic unit of chromatin is the nucleosome, which consists of a short piece of DNA wrapped around a core of eight histone proteins. In addition to their role

  9. Dynamic Conformations of Nucleosome Arrays in Solution from Small-Angle X-ray Scattering

    Energy Technology Data Exchange (ETDEWEB)

    Howell, Steven C. [George Washington Univ., Washington, DC (United States)

    2016-01-31

    We set out to determine quantitative information regarding the dynamic conformation of nucleosome arrays in solution using experimental SAXS. Toward this end, we developed a CG simulation algorithm for dsDNA which rapidly generates ensembles of structures through Metropolis MC sampling of a Markov chain.

  10. Contribution of histone N-terminal tails to the structure and stability of nucleosomes.

    Science.gov (United States)

    Iwasaki, Wakana; Miya, Yuta; Horikoshi, Naoki; Osakabe, Akihisa; Taguchi, Hiroyuki; Tachiwana, Hiroaki; Shibata, Takehiko; Kagawa, Wataru; Kurumizaka, Hitoshi

    2013-01-01

    Histones are the protein components of the nucleosome, which forms the basic architecture of eukaryotic chromatin. Histones H2A, H2B, H3, and H4 are composed of two common regions, the "histone fold" and the "histone tail". Many efforts have been focused on the mechanisms by which the post-translational modifications of histone tails regulate the higher-order chromatin architecture. On the other hand, previous biochemical studies have suggested that histone tails also affect the structure and stability of the nucleosome core particle itself. However, the precise contributions of each histone tail are unclear. In the present study, we determined the crystal structures of four mutant nucleosomes, in which one of the four histones, H2A, H2B, H3, or H4, lacked the N-terminal tail. We found that the deletion of the H2B or H3 N-terminal tail affected histone-DNA interactions and substantially decreased nucleosome stability. These findings provide important information for understanding the complex roles of histone tails in regulating chromatin structure.

  11. In vitro reconstitution and biochemical analyses of the Schizosaccharomyces pombe nucleosome.

    Science.gov (United States)

    Koyama, Masako; Nagakura, Wataru; Tanaka, Hiroki; Kujirai, Tomoya; Chikashige, Yuji; Haraguchi, Tokuko; Hiraoka, Yasushi; Kurumizaka, Hitoshi

    2017-01-22

    Schizosaccharomyces pombe, which has a small genome but shares many physiological functions with higher eukaryotes, is a useful single-cell, model eukaryotic organism. In particular, many features concerning chromatin structure and dynamics, including heterochromatin, centromeres, telomeres, and DNA replication origins, are well conserved between S. pombe and higher eukaryotes. However, the S. pombe nucleosome, the fundamental structural unit of chromatin, has not been reconstituted in vitro. In the present study, we established the method to purify S. pombe histones H2A, H2B, H3, and H4, and successfully reconstituted the S. pombe nucleosome in vitro. Our thermal stability assay and micrococcal nuclease treatment assay revealed that the S. pombe nucleosome is markedly unstable and its DNA ends are quite accessible, as compared to the canonical human nucleosome. These findings are important to understand the mechanisms of epigenetic genomic DNA regulation in fission yeast. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. The p400 ATPase regulates nucleosome stability and chromatin ubiquitination during DNA repair

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    Xu, Ye; Sun, Yingli; Jiang, Xiaofeng; Ayrapetov, Marina K.; Moskwa, Patryk; Yang, Shenghong; Weinstock, David M.

    2010-01-01

    The complexity of chromatin architecture presents a significant barrier to the ability of the DNA repair machinery to access and repair DNA double-strand breaks (DSBs). Consequently, remodeling of the chromatin landscape adjacent to DSBs is vital for efficient DNA repair. Here, we demonstrate that DNA damage destabilizes nucleosomes within chromatin regions that correspond to the γ-H2AX domains surrounding DSBs. This nucleosome destabilization is an active process requiring the ATPase activity of the p400 SWI/SNF ATPase and histone acetylation by the Tip60 acetyltransferase. p400 is recruited to DSBs by a mechanism that is independent of ATM but requires mdc1. Further, the destabilization of nucleosomes by p400 is required for the RNF8-dependent ubiquitination of chromatin, and for the subsequent recruitment of brca1 and 53BP1 to DSBs. These results identify p400 as a novel DNA damage response protein and demonstrate that p400-mediated alterations in nucleosome and chromatin structure promote both chromatin ubiquitination and the accumulation of brca1 and 53BP1 at sites of DNA damage. PMID:20876283

  13. Are liquid crystalline properties of nucleosomes involved in chromosome structure and dynamics?

    Science.gov (United States)

    Livolant, Françoise; Mangenot, Stéphanie; Leforestier, Amélie; Bertin, Aurélie; Frutos, Marta de; Raspaud, Eric; Durand, Dominique

    2006-10-15

    Nucleosome core particles correspond to the structural units of eukaryotic chromatin. They are charged colloids, 101 Angstrom in diameter and 55 Angstrom in length, formed by the coiling of a 146/147 bp DNA fragment (50 nm) around the histone protein octamer. Solutions of these particles can be concentrated, under osmotic pressure, up to the concentrations found in the nuclei of living cells. In the presence of monovalent cations (Na(+)), nucleosomes self-assemble into crystalline or liquid crystalline phases. A lamello-columnar phase is observed at 'low salt' concentrations, while a two-dimensional hexagonal phase and a three-dimensional quasi-hexagonal phase form at 'high salt' concentrations. We followed the formation of these phases from the dilute isotropic solutions to the ordered phases by combining cryoelectron microscopy and X-ray diffraction analyses. The phase diagram is presented as a function of the monovalent salt concentration and applied osmotic pressure. An alternative method to condense nucleosomes is to induce their aggregation upon addition of divalent or multivalent cations (Mg(2+), spermidine(3+) and spermine(4+)). Ordered phases are also found in the aggregates. We also discuss whether these condensed phases of nucleosomes may be relevant from a biological point of view.

  14. Nucleosome exclusion from the interspecies-conserved central AT-rich region of the Ars insulator.

    Science.gov (United States)

    Takagi, Haruna; Inai, Yuta; Watanabe, Shun-ichiro; Tatemoto, Sayuri; Yajima, Mamiko; Akasaka, Koji; Yamamoto, Takashi; Sakamoto, Naoaki

    2012-01-01

    The Ars insulator is a boundary element identified in the upstream region of the arylsulfatase (HpArs) gene in the sea urchin, Hemicentrotus pulcherrimus, and possesses the ability to both block enhancer-promoter communications and protect transgenes from silent chromatin. To understand the molecular mechanism of the Ars insulator, we investigated the correlation between chromatin structure, DNA structure and insulator activity. Nuclease digestion of nuclei isolated from sea urchin embryos revealed the presence of a nuclease-hypersensitive site within the Ars insulator. Analysis of micrococcal nuclease-sensitive sites in the Ars insulator, reconstituted with nucleosomes, showed the exclusion of nucleosomes from the central AT-rich region. Furthermore, the central AT-rich region in naked DNA was sensitive to nucleotide base modification by diethylpyrocarbonate (DEPC). These observations suggest that non-B-DNA structures in the central AT-rich region may inhibit nucleosomal formation, which leads to nuclease hypersensitivity. Furthermore, comparison of nucleotide sequences between the HpArs gene and its ortholog in Strongylocentrotus purpuratus revealed that the central AT-rich region of the Ars insulator is conserved, and this conserved region showed significant enhancer blocking activity. These results suggest that the central AT-rich nucleosome-free region plays an important role in the function of the Ars insulator.

  15. Nucleosome-releasing factor: a new role for factor VII-activating protease (FSAP)

    NARCIS (Netherlands)

    Zeerleder, Sacha; Zwart, Bas; te Velthuis, Henk; Stephan, Femke; Manoe, Rishi; Rensink, Irma; Aarden, Lucien A.

    2008-01-01

    Plasma proteins such as early complement components and IgM are involved in the removal of late apoptotic or secondary necrotic (sn) cells. We have recently described how a plasma protease that could be inhibited by the protease inhibitor aprotinin was essential to remove nucleosomes from sn cells.

  16. TRF1 and TRF2 binding to telomeres is modulated by nucleosomal organization.

    Science.gov (United States)

    Galati, Alessandra; Micheli, Emanuela; Alicata, Claudia; Ingegnere, Tiziano; Cicconi, Alessandro; Pusch, Miriam Caroline; Giraud-Panis, Marie-Josèphe; Gilson, Eric; Cacchione, Stefano

    2015-07-13

    The ends of eukaryotic chromosomes need to be protected from the activation of a DNA damage response that leads the cell to replicative senescence or apoptosis. In mammals, protection is accomplished by a six-factor complex named shelterin, which organizes the terminal TTAGGG repeats in a still ill-defined structure, the telomere. The stable interaction of shelterin with telomeres mainly depends on the binding of two of its components, TRF1 and TRF2, to double-stranded telomeric repeats. Tethering of TRF proteins to telomeres occurs in a chromatin environment characterized by a very compact nucleosomal organization. In this work we show that binding of TRF1 and TRF2 to telomeric sequences is modulated by the histone octamer. By means of in vitro models, we found that TRF2 binding is strongly hampered by the presence of telomeric nucleosomes, whereas TRF1 binds efficiently to telomeric DNA in a nucleosomal context and is able to remodel telomeric nucleosomal arrays. Our results indicate that the different behavior of TRF proteins partly depends on the interaction with histone tails of their divergent N-terminal domains. We propose that the interplay between the histone octamer and TRF proteins plays a role in the steps leading to telomere deprotection. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  17. Chromatin modification by PSC occurs at one PSC per nucleosome and does not require the acidic patch of histone H2A.

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    Stanley M Lo

    Full Text Available Chromatin architecture is regulated through both enzymatic and non-enzymatic activities. For example, the Polycomb Group (PcG proteins maintain developmental gene silencing using an array of chromatin-based mechanisms. The essential Drosophila PcG protein, Posterior Sex Combs (PSC, compacts chromatin and inhibits chromatin remodeling and transcription through a non-enzymatic mechanism involving nucleosome bridging. Nucleosome bridging is achieved through a combination of nucleosome binding and self-interaction. Precisely how PSC interacts with chromatin to bridge nucleosomes is not known and is the subject of this work. We determine the stoichiometry of PSC-chromatin interactions in compact chromatin (in which nucleosomes are bridged using Scanning Transmission Electron Microscopy (STEM. We find that full compaction occurs with one PSC per nucleosome. In addition to compacting chromatin, we show that PSC oligomerizes nucleosome arrays. PSC-mediated oligomerization of chromatin occurs at similar stoichiometry as compaction suggesting it may also involve nucleosome bridging. Interactions between the tail of histone H4 and the acidic patch of histone H2A are important for chromatin folding and oligomerization, and several chromatin proteins bind the histone H2A acidic patch. However, mutation of the acidic patch of histone H2A does not affect PSC's ability to inhibit chromatin remodeling or bridge nucleosomes. In fact, PSC does not require nucleosomes for bridging activity but can bridge naked DNA segments. PSC clusters nucleosomes on sparsely assembled templates, suggesting it interacts preferentially with nucleosomes over bare DNA. This may be due to the ability of PSC to bind free histones. Our data are consistent with a model in which each PSC binds a nucleosome and at least one other PSC to directly bridge nucleosomes and compact chromatin, but also suggest that naked DNA can be included in compacted structures. We discuss how our data

  18. The WD40 Domain of HIRA Is Essential for RI-nucleosome Assembly in Xenopus Egg Extracts.

    Science.gov (United States)

    Zhu, Ruibin; Iwabuchi, Mari; Ohsumi, Keita

    2017-01-01

    Histone chaperones are a group of histone-binding proteins that facilitate the assembly of nucleosomes, the fundamental structural units of chromatin in eukaryotes. In nucleosome assembly, deposition of a histone H3-H4 tetramer onto DNA is the first and critical step, which is mediated by the histone chaperones HIRA and CAF-1. HIRA and CAF-1 are reportedly involved in DNA replication independent (RI) and replication coupled nucleosome assembly, respectively. However, the mechanisms by which they mediate histone deposition remain unclear. In this study, we focused on the mechanism by which HIRA induces RI-nucleosome assembly. We looked for HIRA domains that are required for nucleosome assembly and its localization to chromatin. We used cell-free extracts from Xenopus eggs that carry out RI-nucleosome assembly of plasmid DNA. We confirmed that HIRA formed stable complexes with Asf1, another histone H3-H4 chaperone, and the HIRA-Asf1 complex was solely responsible for RI-nucleosome assembly in egg extracts. We further demonstrated that the HIRA N-terminus containing the WD40 domain, which comprises seven WD40 repeats, and the B domain, to which Asf1 binds, were essential for RI-nucleosome assembly; the three WD40 repeats from the N-terminus were especially critical. Using egg extracts that reproduce nuclear formation accompanying the duplication of chromatin, we also demonstrated that the Hir domain was indispensable for the binding of HIRA to chromatin. Thus, the WD40 and B domains are the core elements for inducing RI-nucleosome assembly. Hir domain regulates the binding to chromatin. Based on these findings, similarities and differences between HIRA and CAF-1 are discussed.

  19. Changes in nucleosome position at transcriptional start sites of specific genes in Zea mays mediator of paramutation1 mutants.

    Science.gov (United States)

    Labonne, Jonathan D J; Dorweiler, Jane E; McGinnis, Karen M

    2013-04-01

    Nucleosomes facilitate compaction of DNA within the confines of the eukaryotic nucleus. This packaging of DNA and histone proteins must accommodate cellular processes, such as transcription and DNA replication. The repositioning of nucleosomes to facilitate cellular processes is likely regulated by several factors. In Zea mays, Mediator of paramutation1 (MOP1) has been demonstrated to be an epigenetic regulator of gene expression. Based on sequence orthology and mutant phenotypes, MOP1 is likely to function in an RNA-dependent pathway to mediate changes to chromatin. High-resolution microarrays were used to assay the distribution of nucleosomes across the transcription start sites (TSSs) of ~400 maize genes in wild type and mutant mop1-1 tissues. Analysis of nucleosome distribution in leaf, immature tassel and ear shoot tissues resulted in the identification of three genes showing consistent differences in nucleosome positioning and occupancy between wild type and mutant mop1-1. These specific changes in nucleosome distribution were located upstream as well as downstream of the TSS. No direct relationship between the specific changes in nucleosome distribution and transcription were observed through quantitative expression analysis in these tissues. In silico prediction suggests that nucleosome positioning is not dictated by intrinsic DNA sequence signals in the TSSs of two of the identified genes, suggesting a role for chromatin remodeling proteins in MOP1-mediated pathways. These results also indicate that MOP1 contributions to nucleosome position may be either separate from changes in gene expression, or cooperative with development and other levels of regulation in coordinating gene expression.

  20. Staphylococcus aureus protease: a probe of exposed, non-basic histone sequences in nucleosomes

    Energy Technology Data Exchange (ETDEWEB)

    Rill, R.L.; Oosterhof, D.K.

    1980-01-01

    The digestion of histones in chicken erythrocyte nucleosome cores and chromatin by Staphylococcus aureus protease was examined. This protease cleaves specifically at acidic residues and prefers glu-X bonds under the conditions used. Only 1 of 24 glutamic and 2 of 13 aspartic acids among all four core histones are located in basic, amino-terminal tails, hence staph. protease is a highly specific probe of exposed non-basic sequences. Staph. protease readily degraded H1, H5, and H3; moderately degraded H2b, and only slightly degraded H2a and H4 in nucleosomes and nucleosome cores. Electrophoresis of core histone fragments from limited digests showed that most glutamic acids were inaccessible, but at least five sites in non-basic sequences were readily cleaved. Tentative assignments of these fragments based on comparisons with products from limited digests of pure histones suggested that most accessible sites in nucleosome cores occur in H3. The most probable sites of H3 cutting are glutamic acids at positions 51, 60, 73, 94, and 97. At least one site in H2b, probably the equivalent of glu-105 in the calf H2b sequence, was accessible. No sites in H2a and H4 appeared highly accessible. H5 was readily cleaved at a site near the amino-terminus. These data substantiate the other evidence that non-basic core histone sequences are located primarily in the nucleosome interior, but that H3 binds to the ends of core DNA and thereby is partly exposed as the upper and lower surfaces of the disk-shaped core.

  1. Thymine dimer formation as a probe of the path of DNA in and between nucleosomes in intact chromatin.

    OpenAIRE

    Pehrson, J R

    1989-01-01

    Photo-induced thymine dimer formation was used to probe nucleosome structure in nuclei. The distribution of thymine dimers in the nucleosome and recent studies of the structure of thymine dimer-containing DNA suggest that the rate of thymine dimer formation is affected by the direction and degree of DNA bending. This premise was used to construct a model of the path of DNA in the nucleosome, which has the following features. (i) There are four regions of sharp bending, two which have been see...

  2. An ensemble of B-DNA dinucleotide geometries lead to characteristic nucleosomal DNA structure and provide plasticity required for gene expression

    Science.gov (United States)

    2011-01-01

    Background A nucleosome is the fundamental repeating unit of the eukaryotic chromosome. It has been shown that the positioning of a majority of nucleosomes is primarily controlled by factors other than the intrinsic preference of the DNA sequence. One of the key questions in this context is the role, if any, that can be played by the variability of nucleosomal DNA structure. Results In this study, we have addressed this question by analysing the variability at the dinucleotide and trinucleotide as well as longer length scales in a dataset of nucleosome X-ray crystal structures. We observe that the nucleosome structure displays remarkable local level structural versatility within the B-DNA family. The nucleosomal DNA also incorporates a large number of kinks. Conclusions Based on our results, we propose that the local and global level versatility of B-DNA structure may be a significant factor modulating the formation of nucleosomes in the vicinity of high-plasticity genes, and in varying the probability of binding by regulatory proteins. Hence, these factors should be incorporated in the prediction algorithms and there may not be a unique 'template' for predicting putative nucleosome sequences. In addition, the multimodal distribution of dinucleotide parameters for some steps and the presence of a large number of kinks in the nucleosomal DNA structure indicate that the linear elastic model, used by several algorithms to predict the energetic cost of nucleosome formation, may lead to incorrect results. PMID:21208404

  3. Reconfiguration of nucleosome-depleted regions at distal regulatory elements accompanies DNA methylation of enhancers and insulators in cancer

    National Research Council Canada - National Science Library

    Taberlay, Phillippa C; Statham, Aaron L; Kelly, Theresa K; Clark, Susan J; Jones, Peter A

    2014-01-01

    .... Here, we evaluate the scope of global epigenetic alterations at enhancers and insulator elements in prostate and breast cancer cells using simultaneous genome-wide mapping of DNA methylation and nucleosome occupancy (NOMe-seq...

  4. Hydroxyl-radical footprinting combined with molecular modeling identifies unique features of DNA conformation and nucleosome positioning.

    Science.gov (United States)

    Shaytan, Alexey K; Xiao, Hua; Armeev, Grigoriy A; Wu, Carl; Landsman, David; Panchenko, Anna R

    2017-09-19

    Nucleosomes are the most abundant protein-DNA complexes in eukaryotes that provide compaction of genomic DNA and are implicated in regulation of transcription, DNA replication and repair. The details of DNA positioning on the nucleosome and the DNA conformation can provide key regulatory signals. Hydroxyl-radical footprinting (HRF) of protein-DNA complexes is a chemical technique that probes nucleosome organization in solution with a high precision unattainable by other methods. In this work we propose an integrative modeling method for constructing high-resolution atomistic models of nucleosomes based on HRF experiments. Our method precisely identifies DNA positioning on nucleosome by combining HRF data for both DNA strands with the pseudo-symmetry constraints. We performed high-resolution HRF for Saccharomyces cerevisiae centromeric nucleosome of unknown structure and characterized it using our integrative modeling approach. Our model provides the basis for further understanding the cooperative engagement and interplay between Cse4p protein and the A-tracts important for centromere function. Published by Oxford University Press on behalf of Nucleic Acids Research 2017.

  5. Dynamic Nucleosome-Depleted Regions at Androgen Receptor Enhancers in the Absence of Ligand in Prostate Cancer Cells ▿

    Science.gov (United States)

    Andreu-Vieyra, Claudia; Lai, John; Berman, Benjamin P.; Frenkel, Baruch; Jia, Li; Jones, Peter A.; Coetzee, Gerhard A.

    2011-01-01

    Nucleosome positioning at transcription start sites is known to regulate gene expression by altering DNA accessibility to transcription factors; however, its role at enhancers is poorly understood. We investigated nucleosome positioning at the androgen receptor (AR) enhancers of TMPRSS2, KLK2, and KLK3/PSA in prostate cancer cells. Surprisingly, a population of enhancer modules in androgen-deprived cultures showed nucleosome-depleted regions (NDRs) in all three loci. Under androgen-deprived conditions, NDRs at the TMPRSS2 enhancer were maintained by the pioneer AR transcriptional collaborator GATA-2. Androgen treatment resulted in AR occupancy, an increased number of enhancer modules with NDRs without changes in footprint width, increased levels of histone H3 acetylation (AcH3), and dimethylation (H3K4me2) at nucleosomes flanking the NDRs. Our data suggest that, in the absence of ligand, AR enhancers exist in an equilibrium in which a percentage of modules are occupied by nucleosomes while others display NDRs. We propose that androgen treatment leads to the disruption of the equilibrium toward a nucleosome-depleted state, rather than to enhancer de novo “remodeling.” This allows the recruitment of histone modifiers, chromatin remodelers, and ultimately gene activation. The “receptive” state described here could help explain AR signaling activation under very low ligand concentrations. PMID:21969603

  6. nucleR: a package for non-parametric nucleosome positioning.

    Science.gov (United States)

    Flores, Oscar; Orozco, Modesto

    2011-08-01

    nucleR is an R/Bioconductor package for a flexible and fast recognition of nucleosome positioning from next generation sequencing and tiling arrays experiments. The software is integrated with standard high-throughput genomics R packages and allows for in situ visualization as well as to export results to common genome browser formats. Additional information and methodological details can be found at http://mmb.pcb.ub.es/nucleR

  7. The HIR corepressor complex binds to nucleosomes generating a distinct protein/DNA complex resistant to remodeling by SWI/SNF

    OpenAIRE

    Prochasson, Philippe; Florens, Laurence; Swanson, Selene K.; Washburn, Michael P.; Workman, Jerry L.

    2005-01-01

    The histone regulatory (HIR) and histone promoter control (HPC) repressor proteins regulate three of the four histone gene loci during the Saccharomyces cerevisiae cell cycle. Here, we demonstrate that Hir1, Hir2, Hir3, and Hpc2 proteins form a stable HIR repressor complex. The HIR complex promotes histone deposition onto DNA in vitro and constitutes a novel nucleosome assembly complex. The HIR complex stably binds to DNA and nucleosomes. Furthermore, HIR complex binding to nucleosomes forms ...

  8. Nucleosome–nucleosome interactions via histone tails and linker DNA regulate nuclear rigidity

    Science.gov (United States)

    Shimamoto, Yuta; Tamura, Sachiko; Masumoto, Hiroshi; Maeshima, Kazuhiro

    2017-01-01

    Cells, as well as the nuclei inside them, experience significant mechanical stress in diverse biological processes, including contraction, migration, and adhesion. The structural stability of nuclei must therefore be maintained in order to protect genome integrity. Despite extensive knowledge on nuclear architecture and components, however, the underlying physical and molecular mechanisms remain largely unknown. We address this by subjecting isolated human cell nuclei to microneedle-based quantitative micromanipulation with a series of biochemical perturbations of the chromatin. We find that the mechanical rigidity of nuclei depends on the continuity of the nucleosomal fiber and interactions between nucleosomes. Disrupting these chromatin features by varying cation concentration, acetylating histone tails, or digesting linker DNA results in loss of nuclear rigidity. In contrast, the levels of key chromatin assembly factors, including cohesin, condensin II, and CTCF, and a major nuclear envelope protein, lamin, are unaffected. Together with in situ evidence using living cells and a simple mechanical model, our findings reveal a chromatin-based regulation of the nuclear mechanical response and provide insight into the significance of local and global chromatin structures, such as those associated with interdigitated or melted nucleosomal fibers. PMID:28428255

  9. A Meier-Gorlin syndrome mutation impairs the ORC1-nucleosome association.

    Science.gov (United States)

    Zhang, Wei; Sankaran, Saumya; Gozani, Or; Song, Jikui

    2015-05-15

    Recent studies have identified several genetic mutations within the BAH domain of human Origin Recognition Complex subunit 1 (hORC1BAH), including the R105Q mutation, implicated in Meier-Gorlin Syndrome (MGS). However, the pathological role of the hORC1 R105Q mutation remains unclear. In this study, we have investigated the interactions of the hORC1BAH domain with histone H4K20me2, DNA, and the nucleosome core particle labeled with H4Kc20me2, a chemical analog of H4K20me2. Our study revealed a nucleosomal DNA binding site for hORC1BAH. The R105Q mutation reduces the hORC1BAH-DNA binding affinity, leading to impaired hORC1BAH-nucleosome interaction, which likely influences DNA replication initiation and MGS pathogenesis. This study provides an etiologic link between the hORC1 R105Q mutation and MGS.

  10. Genome-wide nucleosome map and cytosine methylation levels of an ancient human genome.

    Science.gov (United States)

    Pedersen, Jakob Skou; Valen, Eivind; Velazquez, Amhed M Vargas; Parker, Brian J; Rasmussen, Morten; Lindgreen, Stinus; Lilje, Berit; Tobin, Desmond J; Kelly, Theresa K; Vang, Søren; Andersson, Robin; Jones, Peter A; Hoover, Cindi A; Tikhonov, Alexei; Prokhortchouk, Egor; Rubin, Edward M; Sandelin, Albin; Gilbert, M Thomas P; Krogh, Anders; Willerslev, Eske; Orlando, Ludovic

    2014-03-01

    Epigenetic information is available from contemporary organisms, but is difficult to track back in evolutionary time. Here, we show that genome-wide epigenetic information can be gathered directly from next-generation sequence reads of DNA isolated from ancient remains. Using the genome sequence data generated from hair shafts of a 4000-yr-old Paleo-Eskimo belonging to the Saqqaq culture, we generate the first ancient nucleosome map coupled with a genome-wide survey of cytosine methylation levels. The validity of both nucleosome map and methylation levels were confirmed by the recovery of the expected signals at promoter regions, exon/intron boundaries, and CTCF sites. The top-scoring nucleosome calls revealed distinct DNA positioning biases, attesting to nucleotide-level accuracy. The ancient methylation levels exhibited high conservation over time, clustering closely with modern hair tissues. Using ancient methylation information, we estimated the age at death of the Saqqaq individual and illustrate how epigenetic information can be used to infer ancient gene expression. Similar epigenetic signatures were found in other fossil material, such as 110,000- to 130,000-yr-old bones, supporting the contention that ancient epigenomic information can be reconstructed from a deep past. Our findings lay the foundation for extracting epigenomic information from ancient samples, allowing shifts in epialleles to be tracked through evolutionary time, as well as providing an original window into modern epigenomics.

  11. Histone H2A variants in nucleosomes and chromatin: more or less stable?

    Science.gov (United States)

    Bönisch, Clemens; Hake, Sandra B.

    2012-01-01

    In eukaryotes, DNA is organized together with histones and non-histone proteins into a highly complex nucleoprotein structure called chromatin, with the nucleosome as its monomeric subunit. Various interconnected mechanisms regulate DNA accessibility, including replacement of canonical histones with specialized histone variants. Histone variant incorporation can lead to profound chromatin structure alterations thereby influencing a multitude of biological processes ranging from transcriptional regulation to genome stability. Among core histones, the H2A family exhibits highest sequence divergence, resulting in the largest number of variants known. Strikingly, H2A variants differ mostly in their C-terminus, including the docking domain, strategically placed at the DNA entry/exit site and implicated in interactions with the (H3–H4)2-tetramer within the nucleosome and in the L1 loop, the interaction interface of H2A–H2B dimers. Moreover, the acidic patch, important for internucleosomal contacts and higher-order chromatin structure, is altered between different H2A variants. Consequently, H2A variant incorporation has the potential to strongly regulate DNA organization on several levels resulting in meaningful biological output. Here, we review experimental evidence pinpointing towards outstanding roles of these highly variable regions of H2A family members, docking domain, L1 loop and acidic patch, and close by discussing their influence on nucleosome and higher-order chromatin structure and stability. PMID:23002134

  12. Colorectal carcinoma: nucleosomes, carcinoembryonic antigen and ca 19-9 as apoptotic markers; a comparative study

    Directory of Open Access Journals (Sweden)

    Mahgoub Samir S

    2011-07-01

    Full Text Available Abstract Background Colorectal carcinoma is a common and often fatal disease in which methods of early detection and monitoring are essential. The present study was conducted for measuring serum levels of nucleosomes, carcinoembryonic antigen (CEA and CA 19-9 in patients newly diagnosed with colorectal carcinoma and confirmed by clinicopathological study. Method Thirty subjects were included in the current study: six normal subjects as a control group with mean age (45.6 ± 7.9 and twenty four colorectal carcinoma patients with mean age (46.9 ± 15.6, which were classified pathologically according to the degree of malignant cell differentiation into well differentiated (group I, moderately differentiated (group II and poorly differentiated (group III. Fasting venous blood samples were collected preoperative. Results The results revealed a significant increase in serum level of nucleosomes in patients with poorly differentiated tumors versus patients with well differentiated tumors (p = 0.041. The levels of CEA and CA19-9 showed no significant increase (p = 0.569 and 0.450, respectively. Conclusion In conclusion, serum level of nucleosomes provides a highly sensitive and specific apoptotic marker for colorectal carcinoma.

  13. Nucleosome Density ChIP-Seq Identifies Distinct Chromatin Modification Signatures Associated with MNase Accessibility.

    Science.gov (United States)

    Lorzadeh, Alireza; Bilenky, Misha; Hammond, Colin; Knapp, David J H F; Li, Luolan; Miller, Paul H; Carles, Annaick; Heravi-Moussavi, Alireza; Gakkhar, Sitanshu; Moksa, Michelle; Eaves, Connie J; Hirst, Martin

    2016-11-15

    Nucleosome position, density, and post-translational modification are widely accepted components of mechanisms regulating DNA transcription but still incompletely understood. We present a modified native ChIP-seq method combined with an analytical framework that allows MNase accessibility to be integrated with histone modification profiles. Application of this methodology to the primitive (CD34+) subset of normal human cord blood cells enabled genomic regions enriched in one versus two nucleosomes marked by histone 3 lysine 4 trimethylation (H3K4me3) and/or histone 3 lysine 27 trimethylation (H3K27me3) to be associated with their transcriptional and DNA methylation states. From this analysis, we defined four classes of promoter-specific profiles and demonstrated that a majority of bivalent marked promoters are heterogeneously marked at a single-cell level in this primitive cell type. Interestingly, extension of this approach to human embryonic stem cells revealed an altered relationship between chromatin modification state and nucleosome content at promoters, suggesting developmental stage-specific organization of histone methylation states. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  14. The HIR corepressor complex binds to nucleosomes generating a distinct protein/DNA complex resistant to remodeling by SWI/SNF.

    Science.gov (United States)

    Prochasson, Philippe; Florens, Laurence; Swanson, Selene K; Washburn, Michael P; Workman, Jerry L

    2005-11-01

    The histone regulatory (HIR) and histone promoter control (HPC) repressor proteins regulate three of the four histone gene loci during the Saccharomyces cerevisiae cell cycle. Here, we demonstrate that Hir1, Hir2, Hir3, and Hpc2 proteins form a stable HIR repressor complex. The HIR complex promotes histone deposition onto DNA in vitro and constitutes a novel nucleosome assembly complex. The HIR complex stably binds to DNA and nucleosomes. Furthermore, HIR complex binding to nucleosomes forms a distinct protein/DNA complex resistant to remodeling by SWI/SNF. Thus, the HIR complex is a novel nucleosome assembly complex which functions with SWI/SNF to regulate transcription.

  15. Dependence of the Linker Histone and Chromatin Condensation on the Nucleosome Environment.

    Science.gov (United States)

    Perišić, Ognjen; Schlick, Tamar

    2017-08-24

    The linker histone (LH), an auxiliary protein that can bind to chromatin and interact with the linker DNA to form stem motifs, is a key element of chromatin compaction. By affecting the chromatin condensation level, it also plays an active role in gene expression. However, the presence and variable concentration of LH in chromatin fibers with different DNA linker lengths indicate that its folding and condensation are highly adaptable and dependent on the immediate nucleosome environment. Recent experimental studies revealed that the behavior of LH in mononucleosomes markedly differs from that in small nucleosome arrays, but the associated mechanism is unknown. Here we report a structural analysis of the behavior of LH in mononucleosomes and oligonucleosomes (2-6 nucleosomes) using mesoscale chromatin simulations. We show that the adapted stem configuration heavily depends on the strength of electrostatic interactions between LH and its parental DNA linkers, and that those interactions tend to be asymmetric in small oligonucleosome systems. Namely, LH in oligonucleosomes dominantly interacts with one DNA linker only, as opposed to mononucleosomes where LH has similar interactions with both linkers and forms a highly stable nucleosome stem. Although we show that the LH condensation depends sensitively on the electrostatic interactions with entering and exiting DNA linkers, other interactions, especially by nonparental cores and nonparental linkers, modulate the structural condensation by softening LH and thus making oligonucleosomes more flexible, in comparison to to mono- and dinucleosomes. We also find that the overall LH/chromatin interactions sensitively depend on the linker length because the linker length determines the maximal nucleosome stem length. For mononucleosomes with DNA linkers shorter than LH, LH condenses fully, while for DNA linkers comparable or longer than LH, the LH extension in mononucleosomes strongly follows the length of DNA linkers

  16. Histone-modifying enzymes, histone modifications and histone chaperones in nucleosome assembly: Lessons learned from Rtt109 histone acetyltransferases

    Science.gov (United States)

    Dahlin, Jayme L; Chen, Xiaoyue; Walters, Michael A.; Zhang, Zhiguo

    2015-01-01

    During DNA replication, nucleosomes ahead of replication forks are disassembled to accommodate replication machinery. Following DNA replication, nucleosomes are then reassembled onto replicated DNA using both parental and newly synthesized histones. This process, termed DNA replication-coupled nucleosome assembly (RCNA), is critical for maintaining genome integrity and for the propagation of epigenetic information, dysfunctions of which have been implicated in cancers and aging. In recent years, it has been shown that RCNA is carefully orchestrated by a series of histone modifications, histone chaperones and histone-modifying enzymes. Interestingly, many features of RCNA are also found in processes involving DNA replication-independent nucleosome assembly like histone exchange and gene transcription. In yeast, histone H3 lysine K56 acetylation (H3K56ac) is found in newly synthesized histone H3 and is critical for proper nucleosome assembly and for maintaining genomic stability. The histone acetyltransferase (HAT) regulator of Ty1 transposition 109 (Rtt109) is the sole enzyme responsible for H3K56ac in yeast. Much research has centered on this particular histone modification and histone-modifying enzyme. This Critical Review summarizes much of our current understanding of nucleosome assembly and highlights many important insights learned from studying Rtt109 HATs in fungi. We highlight some seminal features in nucleosome assembly conserved in mammalian systems and describe some of the lingering questions in the field. Further studying fungal and mammalian chromatin assembly may have important public health implications, including deeper understandings of human cancers and aging as well as the pursuit of novel anti-fungal therapies. PMID:25365782

  17. Cell-free DNA Comprises an In Vivo Nucleosome Footprint that Informs Its Tissues-Of-Origin.

    Science.gov (United States)

    Snyder, Matthew W; Kircher, Martin; Hill, Andrew J; Daza, Riza M; Shendure, Jay

    2016-01-14

    Nucleosome positioning varies between cell types. By deep sequencing cell-free DNA (cfDNA), isolated from circulating blood plasma, we generated maps of genome-wide in vivo nucleosome occupancy and found that short cfDNA fragments harbor footprints of transcription factors. The cfDNA nucleosome occupancies correlate well with the nuclear architecture, gene structure, and expression observed in cells, suggesting that they could inform the cell type of origin. Nucleosome spacing inferred from cfDNA in healthy individuals correlates most strongly with epigenetic features of lymphoid and myeloid cells, consistent with hematopoietic cell death as the normal source of cfDNA. We build on this observation to show how nucleosome footprints can be used to infer cell types contributing to cfDNA in pathological states such as cancer. Since this strategy does not rely on genetic differences to distinguish between contributing tissues, it may enable the noninvasive monitoring of a much broader set of clinical conditions than currently possible. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Topological diversity of chromatin fibers: Interplay between nucleosome repeat length, DNA linking number and the level of transcription

    Directory of Open Access Journals (Sweden)

    Davood Norouzi

    2015-11-01

    Full Text Available The spatial organization of nucleosomes in 30-nm fibers remains unknown in detail. To tackle this problem, we analyzed all stereochemically possible configurations of two-start chromatin fibers with DNA linkers L = 10-70 bp (nucleosome repeat length NRL = 157-217 bp. In our model, the energy of a fiber is a sum of the elastic energy of the linker DNA, steric repulsion, electrostatics, and the H4 tail-acidic patch interaction between two stacked nucleosomes. We found two families of energetically feasible conformations of the fibers—one observed earlier, and the other novel. The fibers from the two families are characterized by different DNA linking numbers—that is, they are topologically different. Remarkably, the optimal geometry of a fiber and its topology depend on the linker length: the fibers with linkers L = 10n and 10n + 5 bp have DNA linking numbers per nucleosome DLk >>-1.5 and -1.0, respectively. In other words, the level of DNA supercoiling is directly related to the length of the inter-nucleosome linker in the chromatin fiber (and therefore, to NRL. We hypothesize that this topological polymorphism of chromatin fibers may play a role in the process of transcription, which is known to generate different levels of DNA supercoiling upstream and downstream from RNA polymerase. A genome-wide analysis of the NRL distribution in active and silent yeast genes yielded results consistent with this assumption.

  19. Periodic distribution of a putative nucleosome positioning motif in human, nonhuman primates, and archaea: mutual information analysis.

    Science.gov (United States)

    Sosa, Daniela; Miramontes, Pedro; Li, Wentian; Mireles, Víctor; Bobadilla, Juan R; José, Marco V

    2013-01-01

    Recently, Trifonov's group proposed a 10-mer DNA motif YYYYYRRRRR as a solution of the long-standing problem of sequence-based nucleosome positioning. To test whether this generic decamer represents a biological meaningful signal, we compare the distribution of this motif in primates and Archaea, which are known to contain nucleosomes, and in Eubacteria, which do not possess nucleosomes. The distribution of the motif is analyzed by the mutual information function (MIF) with a shifted version of itself (MIF profile). We found common features in the patterns of this generic decamer on MIF profiles among primate species, and interestingly we found conspicuous but dissimilar MIF profiles for each Archaea tested. The overall MIF profiles for each chromosome in each primate species also follow a similar pattern. Trifonov's generic decamer may be a highly conserved motif for the nucleosome positioning, but we argue that this is not the only motif. The distribution of this generic decamer exhibits previously unidentified periodicities, which are associated to highly repetitive sequences in the genome. Alu repetitive elements contribute to the most fundamental structure of nucleosome positioning in higher Eukaryotes. In some regions of primate chromosomes, the distribution of the decamer shows symmetrical patterns including inverted repeats.

  20. Periodic Distribution of a Putative Nucleosome Positioning Motif in Human, Nonhuman Primates, and Archaea: Mutual Information Analysis

    Directory of Open Access Journals (Sweden)

    Daniela Sosa

    2013-01-01

    Full Text Available Recently, Trifonov's group proposed a 10-mer DNA motif YYYYYRRRRR as a solution of the long-standing problem of sequence-based nucleosome positioning. To test whether this generic decamer represents a biological meaningful signal, we compare the distribution of this motif in primates and Archaea, which are known to contain nucleosomes, and in Eubacteria, which do not possess nucleosomes. The distribution of the motif is analyzed by the mutual information function (MIF with a shifted version of itself (MIF profile. We found common features in the patterns of this generic decamer on MIF profiles among primate species, and interestingly we found conspicuous but dissimilar MIF profiles for each Archaea tested. The overall MIF profiles for each chromosome in each primate species also follow a similar pattern. Trifonov’s generic decamer may be a highly conserved motif for the nucleosome positioning, but we argue that this is not the only motif. The distribution of this generic decamer exhibits previously unidentified periodicities, which are associated to highly repetitive sequences in the genome. Alu repetitive elements contribute to the most fundamental structure of nucleosome positioning in higher Eukaryotes. In some regions of primate chromosomes, the distribution of the decamer shows symmetrical patterns including inverted repeats.

  1. PING 2.0: an R/Bioconductor package for nucleosome positioning using next-generation sequencing data.

    Science.gov (United States)

    Woo, Sangsoon; Zhang, Xuekui; Sauteraud, Renan; Robert, François; Gottardo, Raphael

    2013-08-15

    MNase-Seq and ChIP-Seq have evolved as popular techniques to study chromatin and histone modification. Although many tools have been developed to identify enriched regions, software tools for nucleosome positioning are still limited. We introduce a flexible and powerful open-source R package, PING 2.0, for nucleosome positioning using MNase-Seq data or MNase- or sonicated- ChIP-Seq data combined with either single-end or paired-end sequencing. PING uses a model-based approach, which enables nucleosome predictions even in the presence of low read counts. We illustrate PING using two paired-end datasets from Saccharomyces cerevisiae and compare its performance with nucleR and ChIPseqR. PING 2.0 is available from the Bioconductor website at http://bioconductor.org. It can run on Linux, Mac and Windows.

  2. The disequilibrium of nucleosomes distribution along chromosomes plays a functional and evolutionarily role in regulating gene expression

    KAUST Repository

    Cui, Peng

    2011-08-19

    To further understand the relationship between nucleosome-space occupancy (NO) and global transcriptional activity in mammals, we acquired a set of genome-wide nucleosome distribution and transcriptome data from the mouse cerebrum and testis based on ChIP (H3)-seq and RNA-seq, respectively. We identified a nearly consistent NO patterns among three mouse tissues-cerebrum, testis, and ESCs-and found, through clustering analysis for transcriptional activation, that the NO variations among chromosomes are closely associated with distinct expression levels between house-keeping (HK) genes and tissue-specific (TS) genes. Both TS and HK genes form clusters albeit the obvious majority. This feature implies that NO patterns, i.e. nucleosome binding and clustering, are coupled with gene clustering that may be functionally and evolutionarily conserved in regulating gene expression among different cell types. © 2011 Cui et al.

  3. The disequilibrium of nucleosomes distribution along chromosomes plays a functional and evolutionarily role in regulating gene expression.

    Directory of Open Access Journals (Sweden)

    Peng Cui

    Full Text Available To further understand the relationship between nucleosome-space occupancy (NO and global transcriptional activity in mammals, we acquired a set of genome-wide nucleosome distribution and transcriptome data from the mouse cerebrum and testis based on ChIP (H3-seq and RNA-seq, respectively. We identified a nearly consistent NO patterns among three mouse tissues--cerebrum, testis, and ESCs--and found, through clustering analysis for transcriptional activation, that the NO variations among chromosomes are closely associated with distinct expression levels between house-keeping (HK genes and tissue-specific (TS genes. Both TS and HK genes form clusters albeit the obvious majority. This feature implies that NO patterns, i.e. nucleosome binding and clustering, are coupled with gene clustering that may be functionally and evolutionarily conserved in regulating gene expression among different cell types.

  4. NAP1L1 accelerates activation and decreases pausing to enhance nucleosome remodeling by CSB.

    Science.gov (United States)

    Lee, Ju Yeon; Lake, Robert J; Kirk, Jaewon; Bohr, Vilhelm A; Fan, Hua-Ying; Hohng, Sungchul

    2017-05-05

    Cockayne syndrome protein B (CSB) belongs to the SWI2/SNF2 ATP-dependent chromatin remodeler family, and CSB is the only ATP-dependent chromatin remodeler essential for transcription-coupled nucleotide excision DNA repair. CSB alone remodels nucleosomes ∼10-fold slower than the ACF remodeling complex. Strikingly, NAP1-like histone chaperones interact with CSB and greatly enhance CSB-mediated chromatin remodeling. While chromatin remodeling by CSB and NAP1-like proteins is crucial for efficient transcription-coupled DNA repair, the mechanism by which NAP1-like proteins enhance chromatin remodeling by CSB remains unknown. Here we studied CSB's DNA-binding and nucleosome-remodeling activities at the single molecule level in real time. We also determined how the NAP1L1 chaperone modulates these activities. We found that CSB interacts with DNA in two principle ways: by simple binding and a more complex association that involves gross DNA distortion. Remarkably, NAP1L1 suppresses both these interactions. Additionally, we demonstrate that nucleosome remodeling by CSB consists of three distinct phases: activation, translocation and pausing, similar to ACF. Importantly, we found that NAP1L1 promotes CSB-mediated remodeling by accelerating both activation and translocation. Additionally, NAP1L1 increases CSB processivity by decreasing the pausing probability during translocation. Our study, therefore, uncovers the different steps of CSB-mediated chromatin remodeling that can be regulated by NAP1L1. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  5. The Role of Histone Tails in the Nucleosome: A Computational Study

    Science.gov (United States)

    Erler, Jochen; Zhang, Ruihan; Petridis, Loukas; Cheng, Xiaolin; Smith, Jeremy C.; Langowski, Jörg

    2014-01-01

    Histone tails play an important role in gene transcription and expression. We present here a systematic computational study of the role of histone tails in the nucleosome, using replica exchange molecular dynamics simulations with an implicit solvent model and different well-established force fields. We performed simulations for all four histone tails, H4, H3, H2A, and H2B, isolated and with inclusion of the nucleosome. The results confirm predictions of previous theoretical studies for the secondary structure of the isolated tails but show a strong dependence on the force field used. In the presence of the entire nucleosome for all force fields, the secondary structure of the histone tails is destabilized. Specific contacts are found between charged lysine and arginine residues and DNA phosphate groups and other binding sites in the minor and major DNA grooves. Using cluster analysis, we found a single dominant configuration of binding to DNA for the H4 and H2A histone tails, whereas H3 and H2B show multiple binding configurations with an equal probability. The leading stabilizing contribution for those binding configurations is the attractive interaction between the positively charged lysine and arginine residues and the negatively charged phosphate groups, and thus the resulting charge neutralization. Finally, we present results of molecular dynamics simulations in explicit solvent to confirm our conclusions. Results from both implicit and explicit solvent models show that large portions of the histone tails are not bound to DNA, supporting the complex role of these tails in gene transcription and expression and making them possible candidates for binding sites of transcription factors, enzymes, and other proteins. PMID:25517156

  6. CAG/CTG repeats alter the affinity for the histone core and the positioning of DNA in the nucleosome.

    Science.gov (United States)

    Volle, Catherine B; Delaney, Sarah

    2012-12-11

    Trinucleotide repeats (TNRs) occur throughout the genome, and their expansion has been linked to several neurodegenerative disorders, including Huntington's disease. TNRs have been studied using both oligonucleotides and plasmids; however, less is know about how repetitive DNA responds to genomic packaging. Here, we investigate the behavior of CAG/CTG repeats incorporated into nucleosome core particles, the most basic unit of chromatin packaging. To assess the general interaction between CAG/CTG repeats and the histone core, we determined the efficiency with which various TNR-containing DNA substrates form nucleosomes, revealing that even short CAG/CTG tracts are robust incorporators. However, the presence of the Huntington gene flanking sequence (htt) decreases the rate of incorporation. Enzymatic and chemical probing revealed repositioning of the DNA in the nucleosome as the number of CAG/CTG repeats increased, regardless of the flanking sequence. Notably, the periodicity of the repeat tract remained unchanged as a function of length and is consistently 10.7 bp per helical turn. In contrast, the periodicity of the nonrepetitive flanking sequence varies and is smaller than the repeat tract at ~10.0-10.5 bp per turn. Furthermore, while the CAG/CTG repeats remain as a canonical duplex in the nucleosome, nucleosome formation causes kinking in a secondary repeat tract in the htt gene, comprised of CCG/CGG repeats. This work highlights the innate ability of CAG/CTG repeats to incorporate and to position in nucleosomes and how that behavior is modulated by the htt flanking sequence. In addition, it illuminates the differences in packaging of healthy and diseased length repeat tracts within the genome.

  7. PING 2.0: an R/Bioconductor package for nucleosome positioning using next-generation sequencing data

    OpenAIRE

    Woo, Sangsoon; Zhang, Xuekui; Sauteraud, Renan; Robert, François; Gottardo, Raphael

    2013-01-01

    Summary: MNase-Seq and ChIP-Seq have evolved as popular techniques to study chromatin and histone modification. Although many tools have been developed to identify enriched regions, software tools for nucleosome positioning are still limited. We introduce a flexible and powerful open-source R package, PING 2.0, for nucleosome positioning using MNase-Seq data or MNase– or sonicated– ChIP-Seq data combined with either single-end or paired-end sequencing. PING uses a model-based approach, which ...

  8. Regulation of Nucleosome Stacking and Chromatin Compaction by the Histone H4 N-Terminal Tail-H2A Acidic Patch Interaction.

    Science.gov (United States)

    Chen, Qinming; Yang, Renliang; Korolev, Nikolay; Liu, Chuan Fa; Nordenskiöld, Lars

    2017-06-30

    Chromatin folding and dynamics are critically dependent on nucleosome-nucleosome interactions with important contributions from internucleosome binding of the histone H4 N-terminal tail K16-R23 domain to the surface of the H2A/H2B dimer. The H4 Lys16 plays a pivotal role in this regard. Using in vitro reconstituted 12-mer nucleosome arrays, we have investigated the mechanism of the H4 N-terminal tail in maintaining nucleosome-nucleosome stacking and mediating intra- and inter-array chromatin compaction, with emphasis on the role of K16 and the positive charge region, R17-R23. Analytical ultracentrifugation sedimentation velocity experiments and precipitation assays were employed to analyze effects on chromatin folding and self-association, respectively. Effects on chromatin folding caused by various mutations and modifications at position K16 in the H4 histone were studied. Additionally, using charge-quenching mutations, we characterized the importance of the interaction of the residues within the H4 positive charge region R17-R23 with the H2A acidic patch of the adjacent nucleosome. Furthermore, crosslinking experiments were conducted to establish the proximity of the basic tail region to the acidic patch. Our data indicate that the positive charge and length of the side chain of H4 K16 are important for its access to the adjacent nucleosome in the process of nucleosome-nucleosome stacking and array folding. The location and orientation of the H4 R17-R23 domain on the H2A/H2B dimer surface of the neighboring nucleosome core particle (NCP) in the compacted chromatin fiber were established. The dominance of electrostatic interactions in maintaining intra-array interaction was demonstrated. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Staging Mobilities / Designing Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    2015-01-01

    asks: what are the physical, social, technical, and cultural conditions to the staging of contemporary urban mobilities? The theoretical framing in the Staging mobilities book is applied to four in-depth cases in the accompanying volume Designing mobilities.This book explore how places, sites......In recent years, urban research has taken a ‘mobilities turn’. There has been a developing realisation that mobilities do not ‘just happen.’ Mobilities are carefully and meticulously designed, planned and staged (from above). However, they are equally importantly acted out, performed and lived......, and systems 'hosting' these multiple and complex mobilities are designed and how they are staging these in terms of their physical layout. By analysing specific cases of ‘mobilities design’ related to the four modes of moving; Walk, Bike, Train, and Car, the book uncover important and until now neglected...

  10. Interactions of the CCAAT-binding trimer NF-Y with nucleosomes.

    Science.gov (United States)

    Motta, M C; Caretti, G; Badaracco, G F; Mantovani, R

    1999-01-15

    NF-Y is a sequence-specific evolutionary conserved activator binding to CCAAT boxes with high affinity and specificity. It is a trimer formed by NF-YA and two putative histone-like subunits, NF-YB and NF-YC, showing similarity to histones H2B and H2A, respectively. We investigated the relationships between NF-Y and chromatin using an Artemia franciscana chromatin assembly system with plasmids containing the Major HistoCompatibility complex class II Ea promoter. The NF-Y trimer, but not single subunits, protects the Y box in the presence of reconstituted chromatin, and it can bind the target sequence during and after assembly. Using reconstitution assays with purified chicken histones, we show that NF-Y associates with preformed nucleosomes. Translational analysis of various Ea fragments of identical length in which the CCAAT box is at different positions indicated that the lateral fragment was slightly more prone to NF-Y binding. In competition experiments, NF-Y is able to prevent formation of nucleosomes significantly. These data support the idea that NF-Y is a gene-specific activator with a built-in capacity to interface with chromatin structures.

  11. ATP-independent cooperative binding of yeast Isw1a to bare and nucleosomal DNA.

    Directory of Open Access Journals (Sweden)

    Anne De Cian

    Full Text Available Among chromatin remodeling factors, the ISWI family displays a nucleosome-enhanced ATPase activity coupled to DNA translocation. While these enzymes are known to bind to DNA, their activity has not been fully characterized. Here we use TEM imaging and single molecule manipulation to investigate the interaction between DNA and yeast Isw1a. We show that Isw1a displays a highly cooperative ATP-independent binding to and bridging between DNA segments. Under appropriate tension, rare single nucleation events can sometimes be observed and loop DNA with a regular step. These nucleation events are often followed by binding of successive complexes bridging between nearby DNA segments in a zipper-like fashion, as confirmed by TEM observations. On nucleosomal substrates, we show that the specific ATP-dependent remodeling activity occurs in the context of cooperative Isw1a complexes bridging extranucleosomal DNA. Our results are interpreted in the context of the recently published partial structure of Isw1a and support its acting as a "protein ruler" (with possibly more than one tick.

  12. The Cac2 subunit is essential for productive histone binding and nucleosome assembly in CAF-1

    Energy Technology Data Exchange (ETDEWEB)

    Mattiroli, Francesca; Gu, Yajie; Balsbaugh, Jeremy L.; Ahn, Natalie G.; Luger, Karolin

    2017-04-18

    Nucleosome assembly following DNA replication controls epigenome maintenance and genome integrity. Chromatin assembly factor 1 (CAF-1) is the histone chaperone responsible for histone (H3-H4)2 deposition following DNA synthesis. Structural and functional details for this chaperone complex and its interaction with histones are slowly emerging. Using hydrogen-deuterium exchange coupled to mass spectrometry, combined with in vitro and in vivo mutagenesis studies, we identified the regions involved in the direct interaction between the yeast CAF-1 subunits, and mapped the CAF-1 domains responsible for H3-H4 binding. The large subunit, Cac1 organizes the assembly of CAF-1. Strikingly, H3-H4 binding is mediated by a composite interface, shaped by Cac1-bound Cac2 and the Cac1 acidic region. Cac2 is indispensable for productive histone binding, while deletion of Cac3 has only moderate effects on H3-H4 binding and nucleosome assembly. These results define direct structural roles for yeast CAF-1 subunits and uncover a previously unknown critical function of the middle subunit in CAF-1.

  13. Effects of cytosine modifications on DNA flexibility and nucleosome mechanical stability.

    Science.gov (United States)

    Ngo, Thuy T M; Yoo, Jejoong; Dai, Qing; Zhang, Qiucen; He, Chuan; Aksimentiev, Aleksei; Ha, Taekjip

    2016-02-24

    Cytosine can undergo modifications, forming 5-methylcytosine (5-mC) and its oxidized products 5-hydroxymethylcytosine (5-hmC), 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC). Despite their importance as epigenetic markers and as central players in cellular processes, it is not well understood how these modifications influence physical properties of DNA and chromatin. Here we report a comprehensive survey of the effect of cytosine modifications on DNA flexibility. We find that even a single copy of 5-fC increases DNA flexibility markedly. 5-mC reduces and 5-hmC enhances flexibility, and 5-caC does not have a measurable effect. Molecular dynamics simulations show that these modifications promote or dampen structural fluctuations, likely through competing effects of base polarity and steric hindrance, without changing the average structure. The increase in DNA flexibility increases the mechanical stability of the nucleosome and vice versa, suggesting a gene regulation mechanism where cytosine modifications change the accessibility of nucleosomal DNA through their effects on DNA flexibility.

  14. Groucho-mediated repression may result from a histone deacetylase-dependent increase in nucleosome density.

    Directory of Open Access Journals (Sweden)

    Clint J Winkler

    2010-04-01

    Full Text Available Groucho (Gro is a Drosophila melanogaster transcriptional corepressor that directly interacts with the histone deacetylase Rpd3. Although previous studies suggest that this interaction is required for repression of Gro-responsive reporters in cultured cells, the in vivo significance of this interaction and the mechanism by which it leads to repression remain largely unexplored. In this study, we show that Gro is partially dependent on Rpd3 for repression, supporting the idea that Rpd3-mediated repression is one mode of Gro-mediated repression. We demonstrate that Gro colocalizes with Rpd3 to the chromatin of a target gene and that this is accompanied by the deacetylation of specific lysines within the N-terminal tails of histones H3 and H4. Gro overexpression leads to wing patterning defects and ectopic repression in the wing disc of transcription directed by the vestigial quadrant enhancer. These effects are reversed by the histone deacetylase inhibitors TSA and HC-Toxin and by the reduction of Rpd3 gene dosage. Furthermore, repression of the vestigial quadrant enhancer is accompanied by a Gro-mediated increase in nucleosome density, an effect that is reversed by histone deacetylase inhibitors. We propose a model in which Gro-mediated histone deacetylation results in increased nucleosome density leading to transcriptional repression.

  15. Groucho-mediated repression may result from a histone deacetylase-dependent increase in nucleosome density.

    Science.gov (United States)

    Winkler, Clint J; Ponce, Alberto; Courey, Albert J

    2010-04-13

    Groucho (Gro) is a Drosophila melanogaster transcriptional corepressor that directly interacts with the histone deacetylase Rpd3. Although previous studies suggest that this interaction is required for repression of Gro-responsive reporters in cultured cells, the in vivo significance of this interaction and the mechanism by which it leads to repression remain largely unexplored. In this study, we show that Gro is partially dependent on Rpd3 for repression, supporting the idea that Rpd3-mediated repression is one mode of Gro-mediated repression. We demonstrate that Gro colocalizes with Rpd3 to the chromatin of a target gene and that this is accompanied by the deacetylation of specific lysines within the N-terminal tails of histones H3 and H4. Gro overexpression leads to wing patterning defects and ectopic repression in the wing disc of transcription directed by the vestigial quadrant enhancer. These effects are reversed by the histone deacetylase inhibitors TSA and HC-Toxin and by the reduction of Rpd3 gene dosage. Furthermore, repression of the vestigial quadrant enhancer is accompanied by a Gro-mediated increase in nucleosome density, an effect that is reversed by histone deacetylase inhibitors. We propose a model in which Gro-mediated histone deacetylation results in increased nucleosome density leading to transcriptional repression.

  16. FACT, the Bur kinase pathway, and the histone co-repressor HirC have overlapping nucleosome-related roles in yeast transcription elongation.

    Directory of Open Access Journals (Sweden)

    Jennifer R Stevens

    Full Text Available Gene transcription is constrained by the nucleosomal nature of chromosomal DNA. This nucleosomal barrier is modulated by FACT, a conserved histone-binding heterodimer. FACT mediates transcription-linked nucleosome disassembly and also nucleosome reassembly in the wake of the RNA polymerase II transcription complex, and in this way maintains the repression of 'cryptic' promoters found within some genes. Here we focus on a novel mutant version of the yeast FACT subunit Spt16 that supplies essential Spt16 activities but impairs transcription-linked nucleosome reassembly in dominant fashion. This Spt16 mutant protein also has genetic effects that are recessive, which we used to show that certain Spt16 activities collaborate with histone acetylation and the activities of a Bur-kinase/Spt4-Spt5/Paf1C pathway that facilitate transcription elongation. These collaborating activities were opposed by the actions of Rpd3S, a histone deacetylase that restores a repressive chromatin environment in a transcription-linked manner. Spt16 activity paralleling that of HirC, a co-repressor of histone gene expression, was also found to be opposed by Rpd3S. Our findings suggest that Spt16, the Bur/Spt4-Spt5/Paf1C pathway, and normal histone abundance and/or stoichiometry, in mutually cooperative fashion, facilitate nucleosome disassembly during transcription elongation. The recessive nature of these effects of the mutant Spt16 protein on transcription-linked nucleosome disassembly, contrasted to its dominant negative effect on transcription-linked nucleosome reassembly, indicate that mutant FACT harbouring the mutant Spt16 protein competes poorly with normal FACT at the stage of transcription-linked nucleosome disassembly, but effectively with normal FACT for transcription-linked nucleosome reassembly. This functional difference is consistent with the idea that FACT association with the transcription elongation complex depends on nucleosome disassembly, and that the

  17. Nitrated nucleosome levels and neuropsychiatric events in systemic lupus erythematosus; a multi-center retrospective case-control study

    DEFF Research Database (Denmark)

    Ferreira, Isabel; Croca, Sara; Raimondo, Maria Gabriella

    2017-01-01

    BACKGROUND: In patients with systemic lupus erythematosus (SLE) there is no serological test that will reliably distinguish neuropsychiatric (NP) events due to active SLE from those due to other causes. Previously we showed that serum levels of nitrated nucleosomes (NN) were elevated in a small n...

  18. Vaccination with L. infantum chagasi nucleosomal histones confers protection against new world cutaneous leishmaniasis caused by Leishmania braziliensis.

    Science.gov (United States)

    Carneiro, Marcia W; Santos, Diego M; Fukutani, Kiyoshi F; Clarencio, Jorge; Miranda, Jose Carlos; Brodskyn, Claudia; Barral, Aldina; Barral-Netto, Manoel; Soto, Manuel; de Oliveira, Camila I

    2012-01-01

    Nucleosomal histones are intracellular proteins that are highly conserved among Leishmania species. After parasite destruction or spontaneous lysis, exposure to these proteins elicits a strong host immune response. In the present study, we analyzed the protective capability of Leishmania infantum chagasi nucleosomal histones against L. braziliensis infection using different immunization strategies. BALB/c mice were immunized with either a plasmid DNA cocktail (DNA) containing four Leishmania nucleosomal histones or with the DNA cocktail followed by the corresponding recombinant proteins plus CpG (DNA/Protein). Mice were later challenged with L. braziliensis, in the presence of sand fly saliva. Lesion development, parasite load and the cellular immune response were analyzed five weeks after challenge. Immunization with either DNA alone or with DNA/Protein was able to inhibit lesion development. This finding was highlighted by the absence of infected macrophages in tissue sections. Further, parasite load at the infection site and in the draining lymph nodes was also significantly lower in vaccinated animals. This outcome was associated with increased expression of IFN-γ and down regulation of IL-4 at the infection site. The data presented here demonstrate the potential use of L. infantum chagasi nucleosomal histones as targets for the development of vaccines against infection with L. braziliensis, as shown by the significant inhibition of disease development following a live challenge.

  19. Vaccination with L. infantum chagasi nucleosomal histones confers protection against new world cutaneous leishmaniasis caused by Leishmania braziliensis.

    Directory of Open Access Journals (Sweden)

    Marcia W Carneiro

    Full Text Available BACKGROUND: Nucleosomal histones are intracellular proteins that are highly conserved among Leishmania species. After parasite destruction or spontaneous lysis, exposure to these proteins elicits a strong host immune response. In the present study, we analyzed the protective capability of Leishmania infantum chagasi nucleosomal histones against L. braziliensis infection using different immunization strategies. METHODOLOGY/PRINCIPAL FINDINGS: BALB/c mice were immunized with either a plasmid DNA cocktail (DNA containing four Leishmania nucleosomal histones or with the DNA cocktail followed by the corresponding recombinant proteins plus CpG (DNA/Protein. Mice were later challenged with L. braziliensis, in the presence of sand fly saliva. Lesion development, parasite load and the cellular immune response were analyzed five weeks after challenge. Immunization with either DNA alone or with DNA/Protein was able to inhibit lesion development. This finding was highlighted by the absence of infected macrophages in tissue sections. Further, parasite load at the infection site and in the draining lymph nodes was also significantly lower in vaccinated animals. This outcome was associated with increased expression of IFN-γ and down regulation of IL-4 at the infection site. CONCLUSION: The data presented here demonstrate the potential use of L. infantum chagasi nucleosomal histones as targets for the development of vaccines against infection with L. braziliensis, as shown by the significant inhibition of disease development following a live challenge.

  20. Probing Enhanced Double-Strand Break Formation at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

    Science.gov (United States)

    Banerjee, Samya; Chakraborty, Supratim; Jacinto, Marco Paolo; Paul, Michael D; Balster, Morgan V; Greenberg, Marc M

    2017-01-10

    DNA is rapidly cleaved under mild alkaline conditions at apyrimidinic/apurinic sites, but the half-life is several weeks in phosphate buffer (pH 7.5). However, abasic sites are ∼100-fold more reactive within nucleosome core particles (NCPs). Histone proteins catalyze the strand scission, and at superhelical location 1.5, the histone H4 tail is largely responsible for the accelerated cleavage. The rate constant for strand scission at an abasic site is enhanced further in a nucleosome core particle when it is part of a bistranded lesion containing a proximal strand break. Cleavage of this form results in a highly deleterious double-strand break. This acceleration is dependent upon the position of the abasic lesion in the NCP and its structure. The enhancement in cleavage rate at an apurinic/apyrimidinic site rapidly drops off as the distance between the strand break and abasic site increases and is negligible once the two forms of damage are separated by 7 bp. However, the enhancement of the rate of double-strand break formation increases when the size of the gap is increased from one to two nucleotides. In contrast, the cleavage rate enhancement at 2-deoxyribonolactone within bistranded lesions is more modest, and it is similar in free DNA and nucleosome core particles. We postulate that the enhanced rate of double-strand break formation at bistranded lesions containing apurinic/apyrimidinic sites within nucleosome core particles is a general phenomenon and is due to increased DNA flexibility.

  1. High-Density Nucleosome Occupancy Map of Human Chromosome 9p21–22 Reveals Chromatin Organization of the Type I Interferon Gene Cluster

    Science.gov (United States)

    Freaney, Jonathan E.; Zhang, Quanwei; Yigit, Erbay; Kim, Rebecca; Widom, Jonathan; Wang, Ji-Ping

    2014-01-01

    Genome-wide investigations have dramatically increased our understanding of nucleosome positioning and the role of chromatin in gene regulation, yet some genomic regions have been poorly represented in human nucleosome maps. One such region is represented by human chromosome 9p21–22, which contains the type I interferon gene cluster that includes 16 interferon alpha genes and the single interferon beta, interferon epsilon, and interferon omega genes. A high-density nucleosome mapping strategy was used to generate locus-wide maps of the nucleosome organization of this biomedically important locus at a steady state and during a time course of infection with Sendai virus, an inducer of interferon gene expression. Detailed statistical and computational analysis illustrates that nucleosomes in this locus exhibit preferences for particular dinucleotide and oligomer DNA sequence motifs in vivo, which are similar to those reported for lower eukaryotic nucleosome–DNA interactions. These data were used to visualize the region's chromatin architecture and reveal features that are common to the organization of all the type I interferon genes, indicating a common nucleosome-mediated gene regulatory paradigm. Additionally, this study clarifies aspects of the dynamic changes that occur with the nucleosome occupying the transcriptional start site of the interferon beta gene after virus infection. PMID:24673249

  2. Computer modeling reveals that modifications of the histone tail charges define salt-dependent interaction of the nucleosome core particles.

    Science.gov (United States)

    Yang, Ye; Lyubartsev, Alexander P; Korolev, Nikolay; Nordenskiöld, Lars

    2009-03-18

    Coarse-grained Langevin molecular dynamics computer simulations were conducted for systems that mimic solutions of nucleosome core particles (NCPs). The NCP was modeled as a negatively charged spherical particle representing the complex of DNA and the globular part of the histones combined with attached strings of connected charged beads modeling the histone tails. The size, charge, and distribution of the tails relative to the core were built to match real NCPs. Three models of NCPs were constructed to represent different extents of covalent modification on the histone tails: (nonmodified) recombinant (rNCP), acetylated (aNCP), and acetylated and phosphorylated (paNCP). The simulation cell contained 10 NCPs in a dielectric continuum with explicit mobile counterions and added salt. The NCP-NCP interaction is decisively dependent on the modification state of the histone tails and on salt conditions. Increasing the monovalent salt concentration (KCl) from salt-free to physiological concentration leads to NCP aggregation in solution for rNCP, whereas NCP associates are observed only occasionally in the system of aNCPs. In the presence of divalent salt (Mg(2+)), rNCPs form dense stable aggregates, whereas aNCPs form aggregates less frequently. Aggregates are formed via histone-tail bridging and accumulation of counterions in the regions of NCP-NCP contacts. The paNCPs do not show NCP-NCP interaction upon addition of KCl or in the presence of Mg(2+). Simulations for systems with a gradual substitution of K(+) for Mg(2+), to mimic the Mg(2+) titration of an NCP solution, were performed. The rNCP system showed stronger aggregation that occurred at lower concentrations of added Mg(2+), compared to the aNCP system. Additional molecular dynamics simulations performed with a single NCP in the simulation cell showed that detachment of the tails from the NCP core was modest under a wide range of salt concentrations. This implies that salt-induced tail dissociation of the

  3. Brownian dynamics simulation of the cross-talking effect among modified histones on conformations of nucleosomes

    Science.gov (United States)

    Duan, Zhao-Wen; Li, Wei; Xie, Ping; Dou, Shuo-Xing; Wang, Peng-Ye

    2010-04-01

    Using Brownian dynamics simulation, we studied the effect of histone modifications on conformations of an array of nucleosomes in a segment of chromatin. The simulation demonstrated that the segment of chromatin shows the dynamic behaviour that its conformation can switch between a state with nearly all of the histones being wrapped by DNA and a state with nearly all of the histones being unwrapped by DNA, thus involving the “cross-talking" interactions among the histones. Each state can stay for a sufficiently long time. These conformational states are essential for gene expression or gene silence. The simulation also shows that these conformational states can be inherited by the daughter DNAs during DNA replication, giving a theoretical explanation of the epigenetic phenomenon.

  4. The Drosophila nucleosome remodeling factor NURF is required for Ecdysteroid signaling and metamorphosis.

    Science.gov (United States)

    Badenhorst, Paul; Xiao, Hua; Cherbas, Lucy; Kwon, So Yeon; Voas, Matt; Rebay, Ilaria; Cherbas, Peter; Wu, Carl

    2005-11-01

    Drosophila NURF is an ISWI-containing ATP-dependent chromatin remodeling complex that regulates transcription by catalyzing nucleosome sliding. To determine in vivo gene targets of NURF, we performed whole genome expression analysis on mutants lacking the NURF-specific subunit NURF301. Strikingly, a large set of ecdysone-responsive targets is included among several hundred NURF-regulated genes. Null Nurf301 mutants do not undergo larval to pupal metamorphosis, and also enhance dominant-negative mutations in ecdysone receptor. Moreover, purified NURF binds EcR in an ecdysone-dependent manner, suggesting it is a direct effector of nuclear receptor activity. The conservation of NURF in mammals has broad implications for steroid signaling.

  5. Long-Range Correlations in Genomic DNA: A Signature of the Nucleosomal Structure

    Science.gov (United States)

    Audit, B.; Thermes, C.; Vaillant, C.; D'Aubenton-Carafa, Y.; Muzy, J. F.; Arneodo, A.

    2001-03-01

    We use the ``wavelet transform microscope'' to carry out a comparative statistical analysis of DNA bending profiles and of the corresponding DNA texts. In the three kingdoms, one reveals on both signals a characteristic scale of 100-200 bp that separates two different regimes of power-law correlations (PLC). In the small-scale regime, PLC are observed in eukaryotic, in double-strand DNA viral, and in archaeal genomes, which contrasts with their total absence in the genomes of eubacteria and their viruses. This strongly suggests that small-scale PLC are related to the mechanisms underlying the wrapping of DNA in the nucleosomal structure. We further speculate that the large scale PLC are the signature of the higher-order structure and dynamics of chromatin.

  6. An Assay for Measuring Histone Variant Exchange within Nucleosomes In Vitro.

    Science.gov (United States)

    Laflamme, Liette; Guillemette, Benoit; Gaudreau, Luc

    2017-01-01

    The incorporation of histone variants into specific chromatin regions is a mechanism by which cells can regulate many important biological processes. One such example is H2A.Z, a highly conserved variant of H2A that is incorporated in genomic regulatory regions and contributes to control gene expression. H2A.Z variant exchange involves the removal of H2A-H2B dimers from a preassembled nucleosome and their replacement with H2A.Z-H2B dimers. A specific family of chromatin remodeling complexes, homologous to the yeast Swr1 complex, have been shown to be capable of this histone exchange activity both in vivo and in vitro. Here, we describe an assay to measure the histone H2A.Z exchange activity of recombinant human p400 on immobilized mononucleosomes in vitro. The assay can be adapted to other histone exchange complexes/catalytic subunits purified from any species.

  7. An all-atom model of the chromatin fiber containing linker histones reveals a versatile structure tuned by the nucleosomal repeat length.

    Directory of Open Access Journals (Sweden)

    Hua Wong

    Full Text Available In the nucleus of eukaryotic cells, histone proteins organize the linear genome into a functional and hierarchical architecture. In this paper, we use the crystal structures of the nucleosome core particle, B-DNA and the globular domain of H5 linker histone to build the first all-atom model of compact chromatin fibers. In this 3D jigsaw puzzle, DNA bending is achieved by solving an inverse kinematics problem. Our model is based on recent electron microscopy measurements of reconstituted fiber dimensions. Strikingly, we find that the chromatin fiber containing linker histones is a polymorphic structure. We show that different fiber conformations are obtained by tuning the linker histone orientation at the nucleosomes entry/exit according to the nucleosomal repeat length. We propose that the observed in vivo quantization of nucleosomal repeat length could reflect nature's ability to use the DNA molecule's helical geometry in order to give chromatin versatile topological and mechanical properties.

  8. Mimotopes for lupus-derived anti-DNA and nucleosome-specific autoantibodies selected from random peptide phage display libraries: facts and follies.

    NARCIS (Netherlands)

    Dieker, J.W.C.; Sun, Y.J.; Jacobs, C.W.M.; Putterman, C.; Monestier, M.; Muller, S.; Vlag, J. van der; Berden, J.H.M.

    2005-01-01

    Autoantibodies against chromatin are the most characteristic serological feature in SLE patients. Anti-dsDNA and nucleosome-specific antibodies are associated with glomerulonephritis, the most serious manifestation of SLE. Identification of peptides mimicking conformational epitopes (so-called

  9. Chemical Synthesis of K34-Ubiquitylated H2B for Nucleosome Reconstitution and Single-Particle Cryo-Electron Microscopy Structural Analysis.

    Science.gov (United States)

    Li, Jiabin; He, Qiaoqiao; Liu, Yuntao; Liu, Sanling; Tang, Shan; Li, Chengmin; Sun, Demeng; Li, Xiaorun; Zhou, Min; Zhu, Ping; Bi, Guoqiang; Zhou, Zhenghong; Zheng, Ji-Shen; Tian, Changlin

    2017-01-17

    Post-translational modifications (e.g., ubiquitylation) of histones play important roles in dynamic regulation of chromatin. Histone ubiquitylation has been speculated to directly influence the structure and dynamics of nucleosomes. However, structural information for ubiquitylated nucleosomes is still lacking. Here we report an alternative strategy for total chemical synthesis of homogenous histone H2B-K34-ubiquitylation (H2B-K34Ub) by using acid-cleavable auxiliary-mediated ligation of peptide hydrazides for site-specific ubiquitylation. Synthetic H2B-K34Ub was efficiently incorporated into nucleosomes and further used for single-particle cryo-electron microscopy (cryo-EM) imaging. The cryo-EM structure of the nucleosome containing H2B-K34Ub suggests that two flexible ubiquitin domains protrude between the DNA chains of the nucleosomes. The DNA chains around the H2B-K34 sites shift and provide more space for ubiquitin to protrude. These analyses indicated local and slight structural influences on the nucleosome with ubiquitylation at the H2B-K34 site. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Computer Modeling Reveals that Modifications of the Histone Tail Charges Define Salt-Dependent Interaction of the Nucleosome Core Particles

    OpenAIRE

    Yang, Ye; Lyubartsev, Alexander P.; Korolev, Nikolay; Nordenskiöld, Lars

    2009-01-01

    Coarse-grained Langevin molecular dynamics computer simulations were conducted for systems that mimic solutions of nucleosome core particles (NCPs). The NCP was modeled as a negatively charged spherical particle representing the complex of DNA and the globular part of the histones combined with attached strings of connected charged beads modeling the histone tails. The size, charge, and distribution of the tails relative to the core were built to match real NCPs. Three models of NCPs were con...

  11. The conformational state of the nucleosome entry–exit site modulates TATA box-specific TBP binding

    Science.gov (United States)

    Hieb, Aaron R.; Gansen, Alexander; Böhm, Vera; Langowski, Jörg

    2014-01-01

    The TATA binding protein (TBP) is a critical transcription factor used for nucleating assembly of the RNA polymerase II machinery. TBP binds TATA box elements with high affinity and kinetic stability and in vivo is correlated with high levels of transcription activation. However, since most promoters use less stable TATA-less or TATA-like elements, while also competing with nucleosome occupancy, further mechanistic insight into TBP's DNA binding properties and ability to access chromatin is needed. Using bulk and single-molecule FRET, we find that TBP binds a minimal consensus TATA box as a two-state equilibrium process, showing no evidence for intermediate states. However, upon addition of flanking DNA sequence, we observe non-specific cooperative binding to multiple DNA sites that compete for TATA-box specificity. Thus, we conclude that TBP binding is defined by a branched pathway, wherein TBP initially binds with little sequence specificity and is thermodynamically positioned by its kinetic stability to the TATA box. Furthermore, we observed the real-time access of TBP binding to TATA box DNA located within the DNA entry–exit site of the nucleosome. From these data, we determined salt-dependent changes in the nucleosome conformation regulate TBP's access to the TATA box, where access is highly constrained under physiological conditions, but is alleviated by histone acetylation and TFIIA. PMID:24829456

  12. The conformational state of the nucleosome entry-exit site modulates TATA box-specific TBP binding.

    Science.gov (United States)

    Hieb, Aaron R; Gansen, Alexander; Böhm, Vera; Langowski, Jörg

    2014-07-01

    The TATA binding protein (TBP) is a critical transcription factor used for nucleating assembly of the RNA polymerase II machinery. TBP binds TATA box elements with high affinity and kinetic stability and in vivo is correlated with high levels of transcription activation. However, since most promoters use less stable TATA-less or TATA-like elements, while also competing with nucleosome occupancy, further mechanistic insight into TBP's DNA binding properties and ability to access chromatin is needed. Using bulk and single-molecule FRET, we find that TBP binds a minimal consensus TATA box as a two-state equilibrium process, showing no evidence for intermediate states. However, upon addition of flanking DNA sequence, we observe non-specific cooperative binding to multiple DNA sites that compete for TATA-box specificity. Thus, we conclude that TBP binding is defined by a branched pathway, wherein TBP initially binds with little sequence specificity and is thermodynamically positioned by its kinetic stability to the TATA box. Furthermore, we observed the real-time access of TBP binding to TATA box DNA located within the DNA entry-exit site of the nucleosome. From these data, we determined salt-dependent changes in the nucleosome conformation regulate TBP's access to the TATA box, where access is highly constrained under physiological conditions, but is alleviated by histone acetylation and TFIIA. © The Author(s) 2014. Published by Oxford University Press on behalf of Nucleic Acids Research.

  13. In vivo and in vitro footprinting of nucleosomes and transcriptional activators using an infrared-fluorescence DNA sequencer.

    Science.gov (United States)

    Morohashi, Nobuyuki; Nakajima, Kumiko; Kuwana, Shunsuke; Tachiwana, Hiroaki; Kurumizaka, Hitoshi; Shimizu, Mitsuhiro

    2008-02-01

    The analysis of nucleosome positions and transcription factor binding in chromatin is a central issue for understanding the mechanisms of gene expression in eukaryotes. Here, we have developed a footprinting technique, using multi-cycle primer extension with an infrared-fluorescence DNA sequencer, to analyze chromatin structure in isolated yeast nuclei and transcriptional activator binding in living yeast cells. Using this technique, the binding of the yeast activators Hap1 and Hap2/3/4/5 to their cognate sites was detectable as hypersensitive sites by in vivo UV-photofootprinting, and the locations of nucleosomes in yeast minichromosomes were determined by micrococcal nuclease mapping. We also applied this method to determine the position of the nucleosome in the 5S DNA fragment reconstituted in vitro. This technique allowed us to eliminate the use of radioactive materials and to perform experiments on common benches. Thus, the footprinting procedure established in this study will be useful to researchers studying DNA-protein interactions and chromatin structure in vivo and in vitro.

  14. The Evolution of Aerobic Fermentation in Schizosaccharomyces pombe Was Associated with Regulatory Reprogramming but not Nucleosome Reorganization

    Science.gov (United States)

    Li, Wen-Hsiung

    2011-01-01

    Aerobic fermentation has evolved independently in two yeast lineages, the Saccharomyces cerevisiae and the Schizosaccharomyces pombe lineages. In the S. cerevisiae lineage, the evolution of aerobic fermentation was shown to be associated with transcriptional reprogramming of the genes involved in respiration and was recently suggested to be linked to changes in nucleosome occupancy pattern in the promoter regions of respiration-related genes. In contrast, little is known about the genetic basis for the evolution of aerobic fermentation in the Sch. pombe lineage. In particular, it is not known whether respiration-related genes in Sch. pombe have undergone a transcriptional reprogramming or changes in nucleosome occupancy pattern in their promoter regions. In this study, we compared genome-wide gene expression profiles of Sch. pombe with those of S. cerevisiae and the aerobic respiration yeast Candida albicans. We found that the expression profile of respiration-related genes in Sch. pombe is similar to that of S. cerevisiae, but different from that of C. albicans, suggesting that their transcriptional regulation has been reprogrammed during the evolution of aerobic fermentation. However, we found no significant nucleosome organization change in the promoter of respiration-related gene in Sch. pombe. PMID:21127171

  15. Mobile Learning Using Mobile Phones

    Science.gov (United States)

    Vicente, Paula

    2013-01-01

    The participation in mobile learning programs is conditioned by having/using mobile communication technology. Those who do not have or use such technology cannot participate in mobile learning programs. This study evaluates who are the most likely participants of mobile learning programs by examining the demographic profile and mobile phone usage…

  16. Nucleosomes, Linker DNA, and Linker Histone form a Unique Structural Motif that Directs the Higher-Order Folding and Compaction of Chromatin

    Science.gov (United States)

    Bednar, Jan; Horowitz, Rachel A.; Grigoryev, Sergei A.; Carruthers, Lenny M.; Hansen, Jeffrey C.; Koster, Abraham J.; Woodcock, Christopher L.

    1998-11-01

    The compaction level of arrays of nucleosomes may be understood in terms of the balance between the self-repulsion of DNA (principally linker DNA) and countering factors including the ionic strength and composition of the medium, the highly basic N termini of the core histones, and linker histones. However, the structural principles that come into play during the transition from a loose chain of nucleosomes to a compact 30-nm chromatin fiber have been difficult to establish, and the arrangement of nucleosomes and linker DNA in condensed chromatin fibers has never been fully resolved. Based on images of the solution conformation of native chromatin and fully defined chromatin arrays obtained by electron cryomicroscopy, we report a linker histone-dependent architectural motif beyond the level of the nucleosome core particle that takes the form of a stem-like organization of the entering and exiting linker DNA segments. DNA completes ≈ 1.7 turns on the histone octamer in the presence and absence of linker histone. When linker histone is present, the two linker DNA segments become juxtaposed ≈ 8 nm from the nucleosome center and remain apposed for 3-5 nm before diverging. We propose that this stem motif directs the arrangement of nucleosomes and linker DNA within the chromatin fiber, establishing a unique three-dimensional zigzag folding pattern that is conserved during compaction. Such an arrangement with peripherally arranged nucleosomes and internal linker DNA segments is fully consistent with observations in intact nuclei and also allows dramatic changes in compaction level to occur without a concomitant change in topology.

  17. Nucleosome Core Particle Disassembly and Assembly Kinetics Studied Using Single-Molecule Fluorescence.

    Science.gov (United States)

    Hazan, Noa Plavner; Tomov, Toma E; Tsukanov, Roman; Liber, Miran; Berger, Yaron; Masoud, Rula; Toth, Katalin; Langowski, Joerg; Nir, Eyal

    2015-10-20

    The stability of the nucleosome core particle (NCP) is believed to play a major role in regulation of gene expression. To understand the mechanisms that influence NCP stability, we studied stability and dissociation and association kinetics under different histone protein (NCP) and NaCl concentrations using single-pair Förster resonance energy transfer and alternating laser excitation techniques. The method enables distinction between folded, unfolded, and intermediate NCP states and enables measurements at picomolar to nanomolar NCP concentrations where dissociation and association reactions can be directly observed. We reproduced the previously observed nonmonotonic dependence of NCP stability on NaCl concentration, and we suggest that this rather unexpected behavior is a result of interplay between repulsive and attractive forces within positively charged histones and between the histones and the negatively charged DNA. Higher NaCl concentrations decrease the attractive force between the histone proteins and the DNA but also stabilize H2A/H2B histone dimers, and possibly (H3/H4)2 tetramers. An intermediate state in which one DNA arm is unwrapped, previously observed at high NaCl concentrations, is also explained by this salt-induced stabilization. The strong dependence of NCP stability on ion and histone concentrations, and possibly on other charged macromolecules, may play a role in chromosomal morphology. Copyright © 2015 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Salt-induced conformation and interaction changes of nucleosome core particles.

    Science.gov (United States)

    Mangenot, Stéphanie; Leforestier, Amélie; Vachette, Patrice; Durand, Dominique; Livolant, Françoise

    2002-01-01

    Small angle x-ray scattering was used to follow changes in the conformation and interactions of nucleosome core particles (NCP) as a function of the monovalent salt concentration C(s). The maximal extension (D(max)) of the NCP (145 +/- 3-bp DNA) increases from 137 +/- 5 A to 165 +/- 5 A when C(s) rises from 10 to 50 mM and remains constant with further increases of C(s) up to 200 mM. In view of the very weak increase of the R(g) value in the same C(s) range, we attribute this D(max) variation to tail extension, a proposal confirmed by simulations of the entire I(q) curves, considering an ideal solution of particles with tails either condensed or extended. This tail extension is observed at higher salt values when particles contain longer DNA fragments (165 +/- 10 bp). The maximal extension of the tails always coincides with the screening of repulsive interactions between particles. The second virial coefficient becomes smaller than the hard sphere virial coefficient and eventually becomes negative (net attractive interactions) for NCP(145). Addition of salt simultaneously screens Coulombic repulsive interactions between NCP and Coulombic attractive interactions between tails and DNA inside the NCP. We discuss how the coupling of these two phenomena may be of biological relevance.

  19. X-ray diffraction characterization of the dense phases formed by nucleosome core particles.

    Science.gov (United States)

    Mangenot, Stéphanie; Leforestier, Amélie; Durand, Dominique; Livolant, Françoise

    2003-04-01

    Multiple dense phases of nucleosome core particles (NCPs) were formed in controlled ionic conditions (15-160 mM monovalent salt, no divalent ions), under osmotic pressures ranging from 4.7 x 10(5) to 2.35 x 10(6) Pa. We present here the x-ray diffraction analysis of these phases. In the lamello-columnar phase obtained at low salt concentration (50 mM), NCPs order into either a two-dimensional columnar hexagonal phase or into three-dimensional orthorhombic (quasi-hexagonal) crystals. The lamellar and hexagonal (or quasi-hexagonal) organizations coexist in the intermediate salt range; their demixing requires a long time. For an applied pressure P = 4.7 10(5) Pa, the calculated NCPs concentration ranges from approximately 280 to 320 mg/ml in the lamello-columnar phase to 495 to 585 mg/ml in the three-dimensional orthorhombic phase. These concentrations cover the concentration of the living cell.

  20. H2A and H2B tails are essential to properly reconstitute nucleosome core particles.

    Science.gov (United States)

    Bertin, Aurélie; Durand, Dominique; Renouard, Madalena; Livolant, Françoise; Mangenot, Stéphanie

    2007-11-01

    The conformation of recombinant Nucleosome Core Particles (NCPs) lacking H2A and H2B histone tails (gH2AgH2B) are studied. The migration of these particles in acrylamide native gels is slowed down compared to intact reconstituted NCPs. gH2AgH2B NCPs are also much more sensitive to nuclease digestion than intact NCPs. Small angle X-ray scattering (SAXS) experiments point out that the absence of H2A and H2B tails produces small but significant conformational changes of the octamers conformation (without wrapped DNA), whereas gH2AgH2B NCP conformations are significantly altered. A separation of about 25-30 bp from the core could account for the experimental curves, but other types of DNA superhelix deformation cannot be excluded. The distorted gH2AgH2B octamer may not allow the correct winding of DNA around the core. The absence of the H2A and H2B tails would further prevent the secondary sliding of the DNA around the core and therefore impedes the stabilisation of the particle. Cryo-electron microscopy on the same particles also shows a detachment of DNA portions from the particle core. The effect is even stronger because the vitrification of the samples worsens the instability of gH2AgH2B NCPs.

  1. Structure and phase diagram of nucleosome core particles aggregated by multivalent cations.

    Science.gov (United States)

    Bertin, Aurélie; Mangenot, Stéphanie; Renouard, Madalena; Durand, Dominique; Livolant, Françoise

    2007-11-15

    The degree of compaction of the eukaryotic chromatin in vivo and in vitro is highly sensitive to the ionic environment. We address the question of the effect of multivalent ions on the interactions and mutual organization of the chromatin structural units, the nucleosome core particles (NCPs). Conditions of precipitation of NCPs in the presence of 10 mM Tris buffer and various amounts of either magnesium (Mg(2+)) or spermidine (Spd(3+)) are explored, compared, and discussed in relation to theoretical models. In addition, the structure of the aggregates is analyzed by complementary techniques: freeze-fracture electron microscopy, cryoelectron microscopy, and x-ray diffraction. In Mg(2+)-NCP aggregates, NCPs tend to stack on top of one another to form columns that are not long-range organized. In the presence of Spd(3+), NCPs precipitate to form a dense isotropic phase, a disordered phase of columns, a two-dimensional columnar hexagonal phase, or a three-dimensional crystal. The more ordered phases (two-dimensional or three-dimensional hexagonal) are found close to the precipitation line, where the number of positive charges carried by cations is slightly larger than the number of available negative charges of the NCPs. All ordered phases coexist with the dense isotropic phases. Formation of hexagonal and columnar phases is prevented by an excess of polycations.

  2. Rtt101-Mms1-Mms22 coordinates replication-coupled sister chromatid cohesion and nucleosome assembly.

    Science.gov (United States)

    Zhang, Jingjing; Shi, Di; Li, Xiaoli; Ding, Lin; Tang, Jun; Liu, Cong; Shirahige, Katsuhiko; Cao, Qinhong; Lou, Huiqiang

    2017-08-01

    Two sister chromatids must be held together by a cohesion process from their synthesis during S phase to segregation in anaphase. Despite its pivotal role in accurate chromosome segregation, how cohesion is established remains elusive. Here, we demonstrate that yeast Rtt101-Mms1, Cul4 family E3 ubiquitin ligases are stronger dosage suppressors of loss-of-function eco1 mutants than PCNA The essential cohesion reaction, Eco1-catalyzed Smc3 acetylation is reduced in the absence of Rtt101-Mms1. One of the adaptor subunits, Mms22, associates directly with Eco1. Point mutations (L61D/G63D) in Eco1 that abolish the interaction with Mms22 impair Smc3 acetylation. Importantly, an eco1LGpol30A251V double mutant displays additive Smc3ac reduction. Moreover, Smc3 acetylation and cohesion defects also occur in the mutants of other replication-coupled nucleosome assembly (RCNA) factors upstream or downstream of Rtt101-Mms1, indicating unanticipated cross talk between histone modifications and cohesin acetylation. These data suggest that fork-associated Cul4-Ddb1 E3s, together with PCNA, coordinate chromatin reassembly and cohesion establishment on the newly replicated sister chromatids, which are crucial for maintaining genome and chromosome stability. © 2017 The Authors.

  3. Genetic identification of a network of factors that functionally interact with the nucleosome remodeling ATPase ISWI.

    Directory of Open Access Journals (Sweden)

    Giosalba Burgio

    2008-06-01

    Full Text Available Nucleosome remodeling and covalent modifications of histones play fundamental roles in chromatin structure and function. However, much remains to be learned about how the action of ATP-dependent chromatin remodeling factors and histone-modifying enzymes is coordinated to modulate chromatin organization and transcription. The evolutionarily conserved ATP-dependent chromatin-remodeling factor ISWI plays essential roles in chromosome organization, DNA replication, and transcription regulation. To gain insight into regulation and mechanism of action of ISWI, we conducted an unbiased genetic screen to identify factors with which it interacts in vivo. We found that ISWI interacts with a network of factors that escaped detection in previous biochemical analyses, including the Sin3A gene. The Sin3A protein and the histone deacetylase Rpd3 are part of a conserved histone deacetylase complex involved in transcriptional repression. ISWI and the Sin3A/Rpd3 complex co-localize at specific chromosome domains. Loss of ISWI activity causes a reduction in the binding of the Sin3A/Rpd3 complex to chromatin. Biochemical analysis showed that the ISWI physically interacts with the histone deacetylase activity of the Sin3A/Rpd3 complex. Consistent with these findings, the acetylation of histone H4 is altered when ISWI activity is perturbed in vivo. These findings suggest that ISWI associates with the Sin3A/Rpd3 complex to support its function in vivo.

  4. DNA Replication Is Required for Circadian Clock Function by Regulating Rhythmic Nucleosome Composition.

    Science.gov (United States)

    Liu, Xiao; Dang, Yunkun; Matsu-Ura, Toru; He, Yubo; He, Qun; Hong, Christian I; Liu, Yi

    2017-07-20

    Although the coupling between circadian and cell cycles allows circadian clocks to gate cell division and DNA replication in many organisms, circadian clocks were thought to function independently of cell cycle. Here, we show that DNA replication is required for circadian clock function in Neurospora. Genetic and pharmacological inhibition of DNA replication abolished both overt and molecular rhythmicities by repressing frequency (frq) gene transcription. DNA replication is essential for the rhythmic changes of nucleosome composition at the frq promoter. The FACT complex, known to be involved in histone disassembly/reassembly, is required for clock function and is recruited to the frq promoter in a replication-dependent manner to promote replacement of histone H2A.Z by H2A. Finally, deletion of H2A.Z uncoupled the dependence of the circadian clock on DNA replication. Together, these results establish circadian clock and cell cycle as interdependent coupled oscillators and identify DNA replication as a critical process in the circadian mechanism. Published by Elsevier Inc.

  5. Organization and expression of the Paramecium caudatum gene encoding nucleosome assembly protein 1.

    Science.gov (United States)

    Nishiyama, N; Sawatsubashi, S; Ishida, M; Yamauchi, K

    2001-12-12

    The complete genomic and partial complementary DNAs encoding the ciliate Paramecium caudatum nucleosome assembly protein 1 (NAP1) have been sequenced. The nap1 gene is situated 1.2 kbp from the hemoglobin (hb) gene, with the 3' end of both genes facing each other. The nap1 gene contains no introns, and encodes a protein of 369 amino acid residues with a calculated molecular weight of 42,627. The P. caudatum NAP1 amino acid sequence shares only 23-27% identity with NAP1 amino acid sequences from other eukaryotes. Although the nap1 transcript was detected in the P. caudatum cells at both the logarithmic and stationary phases, its level increased during the stationary phase. Southern blot analysis and polymerase chain reaction amplification revealed that the P. caudatum macronucleus has a heterogeneous composition at genomic regions around the nap1 gene. The present studies indicate the nap1 and hb genes are closely arranged in the macronucleus with the intergenic region between their sequences heterogeneously composed.

  6. Nucleosome dynamics and maintenance of epigenetic states of CpG islands

    Science.gov (United States)

    Sneppen, Kim; Dodd, Ian B.

    2016-06-01

    Methylation of mammalian DNA occurs primarily at CG dinucleotides. These CpG sites are located nonrandomly in the genome, tending to occur within high density clusters of CpGs (islands) or within large regions of low CpG density. Cluster methylation tends to be bimodal, being dominantly unmethylated or mostly methylated. For CpG clusters near promoters, low methylation is associated with transcriptional activity, while high methylation is associated with gene silencing. Alternative CpG methylation states are thought to be stable and heritable, conferring localized epigenetic memory that allows transient signals to create long-lived gene expression states. Positive feedback where methylated CpG sites recruit enzymes that methylate nearby CpGs, can produce heritable bistability but does not easily explain that as clusters increase in size or density they change from being primarily methylated to primarily unmethylated. Here, we show that an interaction between the methylation state of a cluster and its occupancy by nucleosomes provides a mechanism to generate these features and explain genome wide systematics of CpG islands.

  7. Characteristic arrangement of nucleosomes is predictive of chromatin interactions at kilobase resolution.

    Science.gov (United States)

    Zhang, Hui; Li, Feifei; Jia, Yan; Xu, Bingxiang; Zhang, Yiqun; Li, Xiaoli; Zhang, Zhihua

    2017-12-15

    High-throughput chromosome conformation capture (3C) technologies, such as Hi-C, have made it possible to survey 3D genome structure. However, obtaining 3D profiles at kilobase resolution at low cost remains a major challenge. Therefore, we herein present an algorithm for precise identification of chromatin interaction sites at kilobase resolution from MNase-seq data, termed chromatin interaction site detector (CISD), and a CISD-based chromatin loop predictor (CISD_loop) that predicts chromatin-chromatin interactions (CCIs) from low-resolution Hi-C data. We show that the predictions of CISD and CISD_loop overlap closely with chromatin interaction analysis by paired-end tag sequencing (ChIA-PET) anchors and loops, respectively. The validity of CISD/CISD_loop was further supported by a 3C assay at about 5 kb resolution. Finally, we demonstrate that only modest amounts of MNase-seq and Hi-C data are sufficient to achieve ultrahigh resolution CCI maps. Our results suggest that CCIs may result in characteristic nucleosomes arrangement patterns flanking the interaction sites, and our algorithms may facilitate precise and systematic investigations of CCIs on a larger scale than hitherto have been possible. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

  8. Crystal structure and stable property of the cancer-associated heterotypic nucleosome containing CENP-A and H3.3.

    Science.gov (United States)

    Arimura, Yasuhiro; Shirayama, Kazuyoshi; Horikoshi, Naoki; Fujita, Risa; Taguchi, Hiroyuki; Kagawa, Wataru; Fukagawa, Tatsuo; Almouzni, Geneviève; Kurumizaka, Hitoshi

    2014-11-19

    The centromere-specific histone H3 variant, CENP-A, is overexpressed in particular aggressive cancer cells, where it can be mislocalized ectopically in the form of heterotypic nucleosomes containing H3.3. In the present study, we report the crystal structure of the heterotypic CENP-A/H3.3 particle and reveal its "hybrid structure", in which the physical characteristics of CENP-A and H3.3 are conserved independently within the same particle. The CENP-A/H3.3 nucleosome forms an unexpectedly stable structure as compared to the CENP-A nucleosome, and allows the binding of the essential centromeric protein, CENP-C, which is ectopically mislocalized in the chromosomes of CENP-A overexpressing cells.

  9. RNA polymerase II stalling promotes nucleosome occlusion and pTEFb recruitment to drive immortalization by Epstein-Barr virus.

    Directory of Open Access Journals (Sweden)

    Richard D Palermo

    2011-10-01

    Full Text Available Epstein-Barr virus (EBV immortalizes resting B-cells and is a key etiologic agent in the development of numerous cancers. The essential EBV-encoded protein EBNA 2 activates the viral C promoter (Cp producing a message of ~120 kb that is differentially spliced to encode all EBNAs required for immortalization. We have previously shown that EBNA 2-activated transcription is dependent on the activity of the RNA polymerase II (pol II C-terminal domain (CTD kinase pTEFb (CDK9/cyclin T1. We now demonstrate that Cp, in contrast to two shorter EBNA 2-activated viral genes (LMP 1 and 2A, displays high levels of promoter-proximally stalled pol II despite being constitutively active. Consistent with pol II stalling, we detect considerable pausing complex (NELF/DSIF association with Cp. Significantly, we observe substantial Cp-specific pTEFb recruitment that stimulates high-level pol II CTD serine 2 phosphorylation at distal regions (up to +75 kb, promoting elongation. We reveal that Cp-specific pol II accumulation is directed by DNA sequences unfavourable for nucleosome assembly that increase TBP access and pol II recruitment. Stalled pol II then maintains Cp nucleosome depletion. Our data indicate that pTEFb is recruited to Cp by the bromodomain protein Brd4, with polymerase stalling facilitating stable association of pTEFb. The Brd4 inhibitor JQ1 and the pTEFb inhibitors DRB and Flavopiridol significantly reduce Cp, but not LMP1 transcript production indicating that Brd4 and pTEFb are required for Cp transcription. Taken together our data indicate that pol II stalling at Cp promotes transcription of essential immortalizing genes during EBV infection by (i preventing promoter-proximal nucleosome assembly and ii necessitating the recruitment of pTEFb thereby maintaining serine 2 CTD phosphorylation at distal regions.

  10. Drosophila Yemanuclein and HIRA cooperate for de novo assembly of H3.3-containing nucleosomes in the male pronucleus.

    Directory of Open Access Journals (Sweden)

    Guillermo A Orsi

    Full Text Available The differentiation of post-meiotic spermatids in animals is characterized by a unique reorganization of their nuclear architecture and chromatin composition. In many species, the formation of sperm nuclei involves the massive replacement of nucleosomes with protamines, followed by a phase of extreme nuclear compaction. At fertilization, the reconstitution of a nucleosome-based paternal chromatin after the removal of protamines requires the deposition of maternally provided histones before the first round of DNA replication. This process exclusively uses the histone H3 variant H3.3 and constitutes a unique case of genome-wide replication-independent (RI de novo chromatin assembly. We had previously shown that the histone H3.3 chaperone HIRA plays a central role for paternal chromatin assembly in Drosophila. Although several conserved HIRA-interacting proteins have been identified from yeast to human, their conservation in Drosophila, as well as their actual implication in this highly peculiar RI nucleosome assembly process, is an open question. Here, we show that Yemanuclein (YEM, the Drosophila member of the Hpc2/Ubinuclein family, is essential for histone deposition in the male pronucleus. yem loss of function alleles affect male pronucleus formation in a way remarkably similar to Hira mutants and abolish RI paternal chromatin assembly. In addition, we demonstrate that HIRA and YEM proteins interact and are mutually dependent for their targeting to the decondensing male pronucleus. Finally, we show that the alternative ATRX/XNP-dependent H3.3 deposition pathway is not involved in paternal chromatin assembly, thus underlining the specific implication of the HIRA/YEM complex for this essential step of zygote formation.

  11. Residues in the Nucleosome Acidic Patch Regulate Histone Occupancy and Are Important for FACT Binding in Saccharomyces cerevisiae.

    Science.gov (United States)

    Hodges, Amelia J; Gloss, Lisa M; Wyrick, John J

    2017-07-01

    The essential histone chaperone FACT plays a critical role in DNA replication, repair, and transcription, primarily by binding to histone H2A-H2B dimers and regulating their assembly into nucleosomes. While FACT histone chaperone activity has been extensively studied, the exact nature of the H2A and H2B residues important for FACT binding remains controversial. In this study, we characterized the functions of residues in the histone H2A and H2B acidic patch, which is important for binding many chromatin-associated factors. We found that mutations in essential acidic patch residues cause a defect in histone occupancy in yeast, even though most of these histone mutants are expressed normally in yeast and form stable nucleosomes in vitro Instead, we show that two acidic patch residues, H2B L109 and H2A E57, are important for histone binding to FACT in vivo We systematically screened mutants in other H2A and H2B residues previously suspected to be important for FACT binding and confirmed the importance of H2B M62 using an in-vivo FACT-binding assay. Furthermore, we show that, like deletion mutants in FACT subunits, an H2A E57 and H2B M62 double mutant is lethal in yeast. In summary, we show that residues in the nucleosome acidic patch promote histone occupancy and are important for FACT binding to H2A-H2B dimers in yeast. Copyright © 2017 by the Genetics Society of America.

  12. Characterizing and controlling intrinsic biases of lambda exonuclease in nascent strand sequencing reveals phasing between nucleosomes and G-quadruplex motifs around a subset of human replication origins

    DEFF Research Database (Denmark)

    Foulk, M. S.; Urban, J. M.; Casella, Cinzia

    2015-01-01

    Nascent strand sequencing (NS-seq) is used to discover DNA replication origins genome-wide, allowing identification of features for their specification. NS-seq depends on the ability of lambda exonuclease (lambda-exo) to efficiently digest parental DNA while leaving RNA-primer protected nascent...... are not general determinants for origin specification but may play a role for a subset. Interestingly, we observed a periodic spacing of G4 motifs and nucleosomes around the peak summits, suggesting that G4s may position nucleosomes at this subset of origins. Finally, we demonstrate that use of Na+ instead of K...

  13. Ubiquitous human 'master' origins of replication are encoded in the DNA sequence via a local enrichment in nucleosome excluding energy barriers.

    Science.gov (United States)

    Drillon, Guénola; Audit, Benjamin; Argoul, Françoise; Arneodo, Alain

    2015-02-18

    As the elementary building block of eukaryotic chromatin, the nucleosome is at the heart of the compromise between the necessity of compacting DNA in the cell nucleus and the required accessibility to regulatory proteins. The recent availability of genome-wide experimental maps of nucleosome positions for many different organisms and cell types has provided an unprecedented opportunity to elucidate to what extent the DNA sequence conditions the primary structure of chromatin and in turn participates in the chromatin-mediated regulation of nuclear functions, such as gene expression and DNA replication. In this study, we use in vivo and in vitro genome-wide nucleosome occupancy data together with the set of nucleosome-free regions (NFRs) predicted by a physical model of nucleosome formation based on sequence-dependent bending properties of the DNA double-helix, to investigate the role of intrinsic nucleosome occupancy in the regulation of the replication spatio-temporal programme in human. We focus our analysis on the so-called replication U/N-domains that were shown to cover about half of the human genome in the germline (skew-N domains) as well as in embryonic stem cells, somatic and HeLa cells (mean replication timing U-domains). The 'master' origins of replication (MaOris) that border these megabase-sized U/N-domains were found to be specified by a few hundred kb wide regions that are hyper-sensitive to DNase I cleavage, hypomethylated, and enriched in epigenetic marks involved in transcription regulation, the hallmarks of localized open chromatin structures. Here we show that replication U/N-domain borders that are conserved in all considered cell lines have an environment highly enriched in nucleosome-excluding-energy barriers, suggesting that these ubiquitous MaOris have been selected during evolution. In contrast, MaOris that are cell-type-specific are mainly regulated epigenetically and are no longer favoured by a local abundance of intrinsic NFRs encoded in

  14. Mobile payment

    CERN Document Server

    Lerner, Thomas

    2013-01-01

    Paying with mobile devices such as mobile phones or smart phones will expand worldwide in the coming years. This development provides opportunities for various industries (banking, telecommunications, credit card business, manufacturers, suppliers, retail) and for consumers.

  15. Mobile Lexicography

    DEFF Research Database (Denmark)

    Køhler Simonsen, Henrik

    2015-01-01

    Mobile phones are ubiquitous and have completely transformed the way we live, work, learn and conduct our everyday activities. Mobile phones have also changed the way users access lexicographic data. In fact, it can be argued that mobile phones and lexicography are not yet compatible. Modern users...... are already mobile – but lexicography is not yet fully ready for the mobile challenge, mobile users and mobile user situations. The article is based on empirical data from two surveys comprising 10 medical doctors, who were asked to look up five medical substances with the medical dictionary app Medicin...... and that lexicographic innovation is needed. A new type of users, new user situations and new access methods call for new lexicographic solutions, and this article proposes a six-pointed hexagram model, which can be used during dictionary app design to lexicographically calibrate the six dimensions in mobile...

  16. Mobile marketing

    OpenAIRE

    Klečková, Zuzana

    2013-01-01

    The main aim of this thesis was to provide a comprehensive overview of the mobile marketing and analyze selected campaigns of Czech mobile marketing in comparison to world successful campaigns. The research contained studying of available literature about the theme to gain general knowledge about the issue. The theoretical part of the thesis contains predominantly various definitions of mobile marketing and its tools, advantages of these tools and some information about Mobile Marketing Assoc...

  17. Mobile Lexicography

    DEFF Research Database (Denmark)

    Køhler Simonsen, Henrik

    2014-01-01

    Users are already mobile, but the question is to which extent knowledge-based dictionary apps are designed for the mobile user situation. The objective of this article is to analyse the characteristics of the mobile user situation and to look further into the stationary user situation...

  18. Staging Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    and lived as people are “staging themselves” (from below). Staging mobilities is a dynamic process between “being staged” (for example, being stopped at traffic lights) and the “mobile staging” of interacting individuals (negotiating a passage on the pavement). Staging Mobilities is about the fact...

  19. DMS-Seq for In Vivo Genome-wide Mapping of Protein-DNA Interactions and Nucleosome Centers

    Directory of Open Access Journals (Sweden)

    Taichi Umeyama

    2017-10-01

    Full Text Available Protein-DNA interactions provide the basis for chromatin structure and gene regulation. Comprehensive identification of protein-occupied sites is thus vital to an in-depth understanding of genome function. Dimethyl sulfate (DMS is a chemical probe that has long been used to detect footprints of DNA-bound proteins in vitro and in vivo. Here, we describe a genomic footprinting method, dimethyl sulfate sequencing (DMS-seq, which exploits the cell-permeable nature of DMS to obviate the need for nuclear isolation. This feature makes DMS-seq simple in practice and removes the potential risk of protein re-localization during nuclear isolation. DMS-seq successfully detects transcription factors bound to cis-regulatory elements and non-canonical chromatin particles in nucleosome-free regions. Furthermore, an unexpected preference of DMS confers on DMS-seq a unique potential to directly detect nucleosome centers without using genetic manipulation. We expect that DMS-seq will serve as a characteristic method for genome-wide interrogation of in vivo protein-DNA interactions.

  20. Footprinting of mammalian promoters: use of a CpG DNA methyltransferase revealing nucleosome positions at a single molecule level.

    Science.gov (United States)

    Fatemi, Mehrnaz; Pao, Martha M; Jeong, Shinwu; Gal-Yam, Einav Nili; Egger, Gerda; Weisenberger, Daniel J; Jones, Peter A

    2005-11-27

    Promoters are molecular 'modules', which are controlled as individual entities yet are often analyzed by nuclease digestion methodologies which, a priori, destroy this modularity. About 40% of mammalian genes contain CpG islands in their promoters and exonic regions, which are normally unmethylated. We developed a footprinting strategy to map the chromatin structure at unmethylated CpG islands by treatment of isolated nuclei with the CpG-specific DNA methyltransferase SssI (M.SssI), followed by genomic bisulfite sequencing of individual progeny DNA molecules. This gave single molecule resolution over the promoter region and allowed for the physical linkage between binding sites on individual promoter molecules to be maintained. Comparison of the p16 promoters in two human cell lines, J82 and LD419, expressing the p16 gene at 25-fold different levels showed that the two cell lines contain remarkably different, heterogeneously positioned nucleosomes over the promoter region, which were not distinguishable by standard methods using nucleases. Our high resolution approach gives a 'digitized' visualization of each promoter providing information regarding nucleosome occupancy and may be utilized to define transcription factor binding and chromatin remodeling.

  1. Chromatin compaction under mixed salt conditions: opposite effects of sodium and potassium ions on nucleosome array folding.

    Science.gov (United States)

    Allahverdi, Abdollah; Chen, Qinming; Korolev, Nikolay; Nordenskiöld, Lars

    2015-02-17

    It is well known that chromatin structure is highly sensitive to the ionic environment. However, the combined effects of a physiologically relevant mixed ionic environment of K(+), Mg(2+) and Na(+), which are the main cations of the cell cytoplasm, has not been systematically investigated. We studied folding and self-association (aggregation) of recombinant 12-mer nucleosome arrays with 177 bp DNA repeat length in solutions of mixtures of K(+) and Mg(2+) or Na(+) and Mg(2+). In the presence of Mg(2+), the addition of sodium ions promotes folding of array into 30-nm fibres, whereas in mixtures of K(+) and Mg(2+), potassium ions abrogate folding. We found that self-association of nucleosome arrays in mixed salt solutions is synergistically promoted by Mg(2+) and monovalent ions, with sodium being slightly more efficient than potassium in amplifying the self-association. The results highlight the importance of a mixed ionic environment for the compaction of chromatin under physiological conditions and demonstrate the complicated nature of the various factors that determine and regulate chromatin compaction in vivo.

  2. lncRNA-Induced Nucleosome Repositioning Reinforces Transcriptional Repression of rRNA Genes upon Hypotonic Stress

    Directory of Open Access Journals (Sweden)

    Zhongliang Zhao

    2016-03-01

    Full Text Available The activity of rRNA genes (rDNA is regulated by pathways that target the transcription machinery or alter the epigenetic state of rDNA. Previous work has established that downregulation of rRNA synthesis in quiescent cells is accompanied by upregulation of PAPAS, a long noncoding RNA (lncRNA that recruits the histone methyltransferase Suv4-20h2 to rDNA, thus triggering trimethylation of H4K20 (H4K20me3 and chromatin compaction. Here, we show that upregulation of PAPAS in response to hypoosmotic stress does not increase H4K20me3 because of Nedd4-dependent ubiquitinylation and proteasomal degradation of Suv4-20h2. Loss of Suv4-20h2 enables PAPAS to interact with CHD4, a subunit of the chromatin remodeling complex NuRD, which shifts the promoter-bound nucleosome into the transcriptional “off” position. Thus, PAPAS exerts a “stress-tailored” dual function in rDNA silencing, facilitating either Suv4-20h2-dependent chromatin compaction or NuRD-dependent changes in nucleosome positioning.

  3. The histone chaperone complex HIR maintains nucleosome occupancy and counterbalances impaired histone deposition in CAF-1 complex mutants.

    Science.gov (United States)

    Duc, Céline; Benoit, Matthias; Le Goff, Samuel; Simon, Lauriane; Poulet, Axel; Cotterell, Sylviane; Tatout, Christophe; Probst, Aline V

    2015-03-01

    Chromatin organization is essential for coordinated gene expression, genome stability, and inheritance of epigenetic information. The main components involved in chromatin assembly are specific complexes such as Chromatin Assembly Factor 1 (CAF-1) and Histone Regulator (HIR), which deposit histones in a DNA synthesis-dependent or -independent manner, respectively. Here, we characterize the role of the plant orthologs Histone Regulator A (HIRA), Ubinuclein (UBN) and Calcineurin Binding protein 1 (CABIN1), which constitute the HIR complex. Arabidopsis loss-of-function mutants for the various subunits of the complex are viable, but hira mutants show reduced fertility. We show that loss of HIRA reduces extractable histone H3 protein levels and decreases nucleosome occupancy at both actively transcribed genes and heterochromatic regions. Concomitantly, HIRA contributes to maintenance of silencing of pericentromeric repeats and certain transposons. A genetic analysis based on crosses between mutants deficient in subunits of the CAF-1 and HIR complexes showed that simultaneous loss of both the CAF-1 and HIR histone H3 chaperone complexes severely affects plant survival, growth and reproductive development. Our results suggest that HIRA partially rescues impaired histone deposition in fas mutants to preserve nucleosome occupancy, implying plasticity in histone variant interaction and deposition. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

  4. Site-specific Acetylation of Histone H3 Decreases Polymerase β Activity on Nucleosome Core Particles in Vitro.

    Science.gov (United States)

    Rodriguez, Yesenia; Hinz, John M; Laughery, Marian F; Wyrick, John J; Smerdon, Michael J

    2016-05-20

    Histone posttranslational modifications have been associated with changes in chromatin structure necessary for transcription, replication, and DNA repair. Acetylation is one of the most studied and best characterized histone posttranslational modifications, but it is not known if histone acetylation modulates base excision repair of DNA lesions in chromatin. To address this question, we generated nucleosome core particles (NCPs) containing site-specifically acetylated H3K14 or H3K56 and measured repair of uracil and single-nucleotide gaps. We find that H3K56Ac and H3K14Ac do not significantly contribute to removal of uracils by uracil DNA glycosylase regardless of the translational or rotational position of the lesions within NCPs. In repair of single-nucleotide gaps, however, the presence of H3K56Ac or H3K14Ac in NCPs decreases the gap-filling activity of DNA polymerase β near the dyad center, with H3K14Ac exhibiting stronger inhibition. To a lesser extent, H3K56Ac induces a similar effect near the DNA ends. Moreover, using restriction enzyme accessibility, we detect no changes in NCP structure or dynamics between H3K14Ac-NCPs and WT-NCPs containing single-nucleotide gaps. Thus, acetylation at H3K56 and H3K14 in nucleosomes may promote alternative gap-filling pathways by inhibiting DNA polymerase β activity. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  5. Mobile Probes in Mobile Learning

    DEFF Research Database (Denmark)

    Ørngreen, Rikke; Blomhøj, Ulla; Duvaa, Uffe

    In this paper experiences from using mobile probes in educational design of a mobile learning application is presented. The probing process stems from the cultural probe method, and was influenced by qualitative interview and inquiry approaches. In the project, the mobile phone was not only acting...... as an agent for acquiring empirical data (as the situation in hitherto mobile probe settings) but was also the technological medium for which data should say something about (mobile learning). Consequently, not only the content of the data but also the ways in which data was delivered and handled, provided...... a valuable dimension for investigating mobile use. The data was collected at the same time as design activities took place and the collective data was analysed based on user experience goals and cognitive processes from interaction design and mobile learning. The mobile probe increased the knowledge base...

  6. Pre-analytical variables of circulating cell-free nucleosomes containing 5-methylcytosine DNA or histone modification H3K9Me3

    DEFF Research Database (Denmark)

    Rasmussen, Louise; Herzog, Marielle; Rømer, Eva

    2016-01-01

    Aim: To evaluate pre-analytical variables of circulating cell-free nucleosomes containing 5-methylcytosine DNA (5mC) or histone modification H3K9Me3 (H3K9Me3). Materials and methods: Six studies were designed to assess the possible influence of pre-analytical variables. Study 1: influence of stas...

  7. Variant histone H2A.Z is globally localized to the promoters of inactive yeast genes and regulates nucleosome positioning.

    Directory of Open Access Journals (Sweden)

    Benoît Guillemette

    2005-12-01

    Full Text Available H2A.Z is an evolutionary conserved histone variant involved in transcriptional regulation, antisilencing, silencing, and genome stability. The mechanism(s by which H2A.Z regulates these various biological functions remains poorly defined, in part due to the lack of knowledge regarding its physical location along chromosomes and the bearing it has in regulating chromatin structure. Here we mapped H2A.Z across the yeast genome at an approximately 300-bp resolution, using chromatin immunoprecipitation combined with tiling microarrays. We have identified 4,862 small regions--typically one or two nucleosomes wide--decorated with H2A.Z. Those "Z loci" are predominantly found within specific nucleosomes in the promoter of inactive genes all across the genome. Furthermore, we have shown that H2A.Z can regulate nucleosome positioning at the GAL1 promoter. Within HZAD domains, the regions where H2A.Z shows an antisilencing function, H2A.Z is localized in a wider pattern, suggesting that the variant histone regulates a silencing and transcriptional activation via different mechanisms. Our data suggest that the incorporation of H2A.Z into specific promoter-bound nucleosomes configures chromatin structure to poise genes for transcriptional activation. The relevance of these findings to higher eukaryotes is discussed.

  8. Effects of macroH2A and H2A.Z on nucleosome structure and dynamics as elucidated by molecular dynamics simulations

    CERN Document Server

    Bowerman, Samuel

    2015-01-01

    Eukaryotes tune the transcriptional activity of their genome by altering the nucleosome core particle through multiple chemical processes. In particular, replacement of the canonical H2A histone with the variants macroH2A and H2A.Z has been shown to affect DNA accessibility and nucleosome stability; however, the processes by which this occurs remain poorly understood. Here, we elucidate the molecular mechanisms of these variants with an extensive molecular dynamics study of the canonical nucleosome along with three variant-containing structures: H2A.Z, macroH2A, and an H2A mutant with macroH2A-like L1 loops. Simulation results show that variant L1 loops play a pivotal role in stabilizing DNA binding to the octamer through direct interactions, core structural rearrangements, and altered allosteric networks in the nucleosome. All variants influence dynamics; however, macroH2A-like systems have the largest effect on energetics. In addition, we provide a comprehensive analysis of allosteric networks in the nucleo...

  9. Ascending the nucleosome face: recognition and function of structured domains in the histone H2A-H2B dimer.

    Science.gov (United States)

    Wyrick, John J; Kyriss, McKenna N M; Davis, William B

    2012-08-01

    Research over the past decade has greatly expanded our understanding of the nucleosome's role as a dynamic hub that is specifically recognized by many regulatory proteins involved in transcription, silencing, replication, repair, and chromosome segregation. While many of these nucleosome interactions are mediated by post-translational modifications in the disordered histone tails, it is becoming increasingly apparent that structured regions of the nucleosome, including the histone fold domains, are also recognized by numerous regulatory proteins. This review will focus on the recognition of structured domains in the histone H2A-H2B dimer, including the acidic patch, the H2A docking domain, the H2B α3-αC helices, and the HAR/HBR domains, and will survey the known biological functions of histone residues within these domains. Novel post-translational modifications and trans-histone regulatory pathways involving structured regions of the H2A-H2B dimer will be highlighted, along with the role of intrinsic disorder in the recognition of structured nucleosome regions. Copyright © 2012 Elsevier B.V. All rights reserved.

  10. Characterization of Complete Histone Tail Proteoforms Using Differential Ion Mobility Spectrometry

    DEFF Research Database (Denmark)

    Shliaha, Pavel V; Baird, Matthew A; Nielsen, Mogens M

    2017-01-01

    Histone proteins are subject to dynamic post-translational modifications (PTMs) that cooperatively modulate the chromatin structure and function. Nearly all functional PTMs are found on the N-terminal histone domains (tails) of ∼50 residues protruding from the nucleosome core. Using high-definition...... differential ion mobility spectrometry (FAIMS) with electron transfer dissociation, we demonstrate rapid baseline gas-phase separation and identification of tails involving monomethylation, trimethylation, acetylation, or phosphorylation in biologically relevant positions. These are by far the largest variant...

  11. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix

    Contemporary society is marked and defined by the ways in which mobile goods, bodies, vehicles, objects, and data are organized, moved and staged. On the background of the ‘mobilities turn’ (e.g. Cresswell 2006, Urry 2007) this book articulates a new and emerging research field, namely that of ‘m......Contemporary society is marked and defined by the ways in which mobile goods, bodies, vehicles, objects, and data are organized, moved and staged. On the background of the ‘mobilities turn’ (e.g. Cresswell 2006, Urry 2007) this book articulates a new and emerging research field, namely...... that of ‘mobilities design’. The book revolves around the following research question: How are design decisions and interventions staging mobilities? It builds upon the Staging Mobilities model (Jensen 2013) in an explorative inquiry into the problems and potentials of the design of mobilities. The exchange value...... between mobilities and design research is twofold. To mobilities research this means getting closer to the ‘material’, and to engage in the creative, explorative and experimental approaches of the design world which offer new potentials for innovative research. Design research, on the other hand, might...

  12. Predictive and prognostic value of circulating nucleosomes and serum biomarkers in patients with metastasized colorectal cancer undergoing Selective Internal Radiation Therapy

    Directory of Open Access Journals (Sweden)

    Fahmueller Yvonne

    2012-01-01

    Full Text Available Abstract Background Selective Internal Radiation Therapy (SIRT is a new and effective locoregional anticancer therapy for colorectal cancer patients with liver metastases. Markers for prediction of therapy response and prognosis are needed for the individual management of those patients undergoing SIRT. Methods Blood samples were prospectively and consecutively taken from 49 colorectal cancer patients with extensive hepatic metastases before, three, six, 24 and 48 h after SIRT to analyze the concentrations of nucleosomes and further laboratory parameters, and to compare them with the response to therapy regularly determined 3 months after therapy and with overall survival. Results Circulating nucleosomes, cytokeratin-19 fragments (CYFRA 21-1, carcinoembryonic antigen (CEA, C-reactive protein (CRP and various liver markers increased already 24 h after SIRT. Pretherapeutical levels of CYFRA 21-1, CEA, cancer antigen 19-9 (CA 19-9, asparate-aminotransferase (AST and lactate dehydrogenase (LDH as well as 24 h values of nucleosomes were significantly higher in patients suffering from disease progression (N = 35 than in non-progressive patients (N = 14. Concerning overall survival, CEA, CA 19-9, CYFRA 21-1, CRP, LDH, AST, choline esterase (CHE, gamma-glutamyl-transferase, alkaline phosphatase, and amylase (all 0 h, 24 h and nucleosomes (24 h were found to be prognostic relevant markers in univariate analyses. In multivariate Cox-Regression analysis, the best prognostic model was obtained for the combination of CRP and AST. When 24 h values were additionally included, nucleosomes (24 h further improved the existing model. Conclusion Panels of biochemical markers are helpful to stratify pretherapeutically colorectal cancer patients for SIR-therapy and to early estimate the response to SIR-therapy.

  13. Genome-Wide Mapping Targets of the Metazoan Chromatin Remodeling Factor NURF Reveals Nucleosome Remodeling at Enhancers, Core Promoters and Gene Insulators.

    Directory of Open Access Journals (Sweden)

    So Yeon Kwon

    2016-04-01

    Full Text Available NURF is a conserved higher eukaryotic ISWI-containing chromatin remodeling complex that catalyzes ATP-dependent nucleosome sliding. By sliding nucleosomes, NURF is able to alter chromatin dynamics to control transcription and genome organization. Previous biochemical and genetic analysis of the specificity-subunit of Drosophila NURF (Nurf301/Enhancer of Bithorax (E(bx has defined NURF as a critical regulator of homeotic, heat-shock and steroid-responsive gene transcription. It has been speculated that NURF controls pathway specific transcription by co-operating with sequence-specific transcription factors to remodel chromatin at dedicated enhancers. However, conclusive in vivo demonstration of this is lacking and precise regulatory elements targeted by NURF are poorly defined. To address this, we have generated a comprehensive map of in vivo NURF activity, using MNase-sequencing to determine at base pair resolution NURF target nucleosomes, and ChIP-sequencing to define sites of NURF recruitment. Our data show that, besides anticipated roles at enhancers, NURF interacts physically and functionally with the TRF2/DREF basal transcription factor to organize nucleosomes downstream of active promoters. Moreover, we detect NURF remodeling and recruitment at distal insulator sites, where NURF functionally interacts with and co-localizes with DREF and insulator proteins including CP190 to establish nucleosome-depleted domains. This insulator function of NURF is most apparent at subclasses of insulators that mark the boundaries of chromatin domains, where multiple insulator proteins co-associate. By visualizing the complete repertoire of in vivo NURF chromatin targets, our data provide new insights into how chromatin remodeling can control genome organization and regulatory interactions.

  14. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix

    2016-01-01

    of increasing concern and societal importance, regardless if the focus is on new inequalities, environmental sustainability, or the meaning of vernacular mobilities design to the everyday life. The theme of ‘mobilities design’ opens up the agenda of architectural research into infrastructure spaces as ‘spaces...... of the perspective in the direction of a material and design oriented turn. In order to fulfill this purpose we articulate a new and emerging research field, namely that of ‘mobilities design’. In our understanding time has come to articulate ‘Mobilities Design’ as a dedicated research field in and of its own....... There is a need for research targeting the material, physical and design-oriented dimensions of the multiple mobilities from the local to the global. Despite its cross-disciplinary identity the ‘mobilities turn’ has not sufficiently capitalized from the potential in exploring issues of material design...

  15. Mobility Divides

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    Contemporary mobilities are cultural and social manifestations, and the mobile practices in the everyday life of billions of humans are re-configuring senses of place, self, other and relationships to the built environment. The way ‘mobile situations’ are staged in designed and built environments......’ in the everyday life and cast light on how design and ‘materialities of mobilites’ are creating differential mobilities across societies, social networks, and communities of practices....... are increasingly becoming ‘second nature’ but also expressions of power, exclusion, and difference. In this talk I will be applying a perspective of ‘mobile situationism’ illustrating how mobile everyday life practices are staged ‘from above’ in planning and policy frameworks, design codes and architectural...

  16. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix; Wind, Simon

    2016-01-01

    In this paper, we identify the nexus between design (architecture, urban design, service design, etc.) and mobilities as a new and emerging research field. In this paper, we apply a “situational mobilities” perspective and take point of departure in the pragmatist question: “What design decisions...... and interventions affords this particular mobile situation?” The paper presents the contours of an emerging research agenda within mobilities research. The advent of “mobilities design” as an emerging research field points towards a critical interest in the material as well as practical consequences of contemporary......-making. The paper proposes that increased understanding of the material affordances facilitated through design provides important insight to planning and policymaking that at times might be in risk of becoming too detached from the everyday life of the mobile subject within contemporary mobilities landscapes....

  17. Mobilities Design

    DEFF Research Database (Denmark)

    Lanng, Ditte Bendix; Wind, Simon; Jensen, Ole B.

    2017-01-01

    is a relevant field of knowledge for urban design, because of its focus on a nuanced conceptualization of the daily journeys that we all undertake in mobilities spaces. Second, the mobilities turn aids a theoretical “mobilization” of the design of mobilities spaces with a point of departure in a fused theory...... field with particular attention to actor-network theory (ANT), which offers tools to understand the embeddedness of mobilities spaces in hybrid and dynamic relationships. Through these linkages between the mobilities turn and urban design, the article suggests a pathway for a carefully radical...... for urban designers and architects to address mobilities spaces in relation not only to technical demands, but also to the wide host of social, cultural, political, economic, and affective formations in which they are embedded and which they influence....

  18. Mobility Work

    DEFF Research Database (Denmark)

    Bardram, Jakob Eyvind; Bossen, Claus

    2005-01-01

    We posit the concept of Mobility Work to describe efforts of moving about people and things as part of accomplishing tasks. Mobility work can be seen as a spatial parallel to the concept of articulation work proposed by the sociologist Anselm Strauss. Articulation work describes efforts...... of coordination necessary in cooperative work, but focuses, we argue, mainly on the temporal aspects of cooperative work. As a supplement, the concept of mobility work focuses on the spatial aspects of cooperative work. Whereas actors seek to diminish the amount of articulation work needed in collaboration...... by constructing Standard Operation Procedures (SOPs), actors minimise mobility work by constructing Standard Operation Configurations (SOCs). We apply the concept of mobility work to the ethnography of hospital work, and argue that mobility arises because of the need to get access to people, places, knowledge and...

  19. Mobile Semiotics

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    2013-01-01

    is a ‘mobile sense making’ where signs and materially situated meanings connect to the moving human body and thus create particular challenges and complexities of making sense of the world. The chapter includes notions of mobility systems and socio-technical networks in order to show how a ‘semiotic layer’ may......This chapter aims to understand the mobile condition of contemporary life with a particular view to the signifying dimension of the environment and its ‘readability’. The chapter explores the potentials of semiotics and its relationship to the new mobilities literature. What takes place...

  20. Extreme heterogeneous composition of the Paramecium caudatum macronuclear genomic DNA between hemoglobin and nucleosome assembly protein-1 genes.

    Science.gov (United States)

    Nishiyama, Norihito; Mikami, Kazuyuki; Matsuoka, Ariki; Ochiai, Takehiko; Yamauchi, Kiyoshi

    2010-02-01

    The intergenic region between the hemoglobin (hb) and nucleosome assembly protein-1 (nap-1) genes in the Paramecium caudatum macronuclear genome was previously found to be heterogeneously composed. Cloning of this intergenic region from the macronuclear genomic DNA identified four unique DNA fragments of different sizes. Sequencing of the cloned fragments revealed extreme heterogeneity and characteristics of both internal eliminated sequence (IES) and imprecise internal deletion sequences (IIDSs) in the intergenic region. Missing sequences were an AT-rich and direct repeats existed in their boundaries. Southern blotting of the total genomic DNA and polymerase chain reaction (PCR) of the total genomic DNAs indicated that there exist a dozen DNA fragments of different sizes in this intergenic region. It is likely that the heterogeneity found in the P. caudatum macronuclear genome results from the variable removal of an intergenic region.

  1. Mobile Semiotics - signs and mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    This paper is about how to comprehend the mobile condition of contemporary life with a particular view to the signifying dimension of the environment and its ‘readability’. The paper explores the potentials of semiotics and its relationship to the new mobilities literature. The theoretical scope...... is therefore an attempt to mobilize semiotics by drawing on a central body of theory within and adjacent to the discipline. For instance the founding works of C. S. Peirce will be related to the contemporary notions of ‘geosemiotics’ by Scollon & Scollon. The paper’s theoretical claim is that semiotics hold...... a potential for mobilities studies if the awareness of seeing the environment as a semiotic layer and system can be sensitized to the insights of the ‘mobilities turn’. Empirically the paper tentatively explores the usefulness of a mobile semiotics approach to cases such as street signage, airport design...

  2. Mobile phones and mobile communication

    DEFF Research Database (Denmark)

    Ling, Richard; Donner, Jonathan

    With staggering swiftness, the mobile phone has become a fixture of daily life in almost every society on earth. In 2007, the world had over 3 billion mobile subscriptions. Prosperous nations boast of having more subscriptions than people. In the developing world, hundreds of millions of people who...... could never afford a landline telephone now have a mobile number of their own. With a mobile in our hand many of us feel safer, more productive, and more connected to loved ones, but perhaps also more distracted and less involved with things happening immediately around us. Written by two leading...... researchers in the field, this volume presents an overview of the mobile telephone as a social and cultural phenomenon. Research is summarized and made accessible though detailed descriptions of ten mobile users from around the world. These illustrate popular debates, as well as deeper social forces at work...

  3. The nucleosome (histone-DNA complex is the TLR9-specific immunostimulatory component of Plasmodium falciparum that activates DCs.

    Directory of Open Access Journals (Sweden)

    Nagaraj M Gowda

    Full Text Available The systemic clinical symptoms of Plasmodium falciparum infection such as fever and chills correspond to the proinflammatory cytokines produced in response to the parasite components released during the synchronized rupture of schizonts. We recently demonstrated that, among the schizont-released products, merozoites are the predominant components that activate dendritic cells (DCs by TLR9-specific recognition to induce the maturation of cells and to produce proinflammatory cytokines. We also demonstrated that DNA is the active constituent and that formation of a DNA-protein complex is essential for the entry of parasite DNA into cells for recognition by TLR9. However, the nature of endogenous protein-DNA complex in the parasite is not known. In this study, we show that parasite nucleosome constitute the major protein-DNA complex involved in the activation of DCs by parasite nuclear material. The parasite components were fractionated into the nuclear and non-nuclear materials. The nuclear material was further fractionated into chromatin and the proteins loosely bound to chromatin. Polynucleosomes and oligonucleosomes were prepared from the chromatin. These were tested for their ability to activate DCs obtained by the FLT3 ligand differentiation of bone marrow cells from the wild type, and TLR2(-/-, TLR9(-/- and MyD88(-/- mice. DCs stimulated with the nuclear material and polynucleosomes as well as mono- and oligonucleosomes efficiently induced the production of proinflammatory cytokines in a TLR9-dependent manner, demonstrating that nucleosomes (histone-DNA complex represent the major TLR9-specific DC-immunostimulatory component of the malaria parasite nuclear material. Thus, our data provide a significant insight into the activation of DCs by malaria parasites and have important implications for malaria vaccine development.

  4. Resource Mobilization

    International Development Research Centre (IDRC) Digital Library (Canada)

    This practical guidebook supports and promotes new and creative thinking on resource mobilization for ... sources and come up with creative resource mobilization strategies to ensure survival. The Importance of ...... funds had already been earmarked for buying new classroom facilities. Maintain integrity of auction. 2.

  5. Mobility Challenges

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lassen, Claus

    2011-01-01

    This article takes point of departure in the challenges to understand the importance of contemporary mobility. The approach advocated is a cross-disciplinary one drawing on sociology, geography, urban planning and design, and cultural studies. As such the perspective is to be seen as a part...... mobilities. In particular the article discusses 1) the physical city, its infrastructures and technological hardware/software, 2) policies and planning strategies for urban mobility and 3) the lived everyday life in the city and the region....... of the so-called ‘mobility turn’ within social science. The perspective is illustrative for the research efforts at the Centre for Mobility and Urban Studies (C-MUS), Aalborg University. The article presents the contours of a theoretical perspective meeting the challenges to research into contemporary urban...

  6. Intensive mobilities

    DEFF Research Database (Denmark)

    Vannini, Phillip; Bissell, David; Jensen, Ole B.

    which relate to transport, housing and employment. Yet we argue that the experiential dimensions of long distance mobilities have not received the attention that they deserve within geographical research on mobilities. This paper combines ideas from mobilities research and contemporary social theory......This paper explores the intensities of long distance commuting journeys as a way of exploring how bodily sensibilities are being changed by the mobilities that they undertake. The context of this paper is that many people are travelling further to work than ever before owing to a variety of factors...... with fieldwork conducted in Canada, Denmark and Australia to develop our understanding of the experiential politics of long distance workers. Rather than focusing on the extensive dimensions of mobilities that are implicated in patterns and trends, our paper turns to the intensive dimensions of this experience...

  7. Mobilities Design

    DEFF Research Database (Denmark)

    Jensen, Ole B.; Lanng, Ditte Bendix

    2016-01-01

    and physical form. The exchange value with design is twofold; first this means getting closer to the ‘material’ which is needed if mobilities research can claim to have understood contemporary mobilities, second it means that the creative, explorative and experimental approaches of the design world becomes...... insights, concepts of space and place, and relations between fixities and flows. The new and emerging field of ‘mobilities design’ will be exploring the borderlines between architecture, urban design, urban planning, and infrastructure design. The field will address the ‘gap’ in research on an issue...... of increasing concern and societal importance, regardless if the focus is on new inequalities, environmental sustainability, or the meaning of vernacular mobilities design to the everyday life. The theme of ‘mobilities design’ opens up the agenda of architectural research into infrastructure spaces as ‘spaces...

  8. Mobile Clouds

    DEFF Research Database (Denmark)

    Fitzek, Frank; Katz, Marcos

    networks, creating a vast fertile ground for novel developments in both research and practical applications Considers research directions, emerging trends and visions This book is an excellent resource for wireless/networking researchers in industry and academia, students and mobile phone programmers...... users in very different ways and for various purposes. The book provides many stimulating examples of resource-sharing applications. Enabling technologies for mobile clouds are also discussed, highlighting the key role of network coding. Mobile clouds have the potential to enhance communications...... examples of mobile clouds applications, based on both existing commercial initiatives as well as proof-of-concept test-beds. Visions and prospects are also discussed, paving the way for further development. As mobile networks and social networks become more and more reliant on each other, the concept...

  9. Sustainable Mobility

    DEFF Research Database (Denmark)

    Kjærulff, Aslak Aamot

    This paper combines strands of mobilities theory and planning theory, and develops a qualitative approach to look across emerging planning practices. By actively following 8 Danish urban and transport planners, over the course of 2 years, we learn how their practices have changed, inspired...... by mobility management, a concept aiming to reduce carbon emissions from transportation in western societies. The article focuses on how municipal planners formulate the role of mobility management activities organized around private companies, and how their practices are connected to wider ideas on planning....

  10. Going Mobile?

    DEFF Research Database (Denmark)

    Tallon, Loic; Froes, Isabel Cristina G.

    2011-01-01

    If the future is mobile, how is the museum community developing within that future? What are the challenges museums face within it? In which directions should we be seeking to evolve our collective knowledge share? It was to gain observations on questions such as these that the 2011 Museums...... & Mobile survey was developed: 660 museum professionals responded. In this paper the authors highlight nine survey observations that they believe are important to the museum community’s increased understanding of and continued progress within mobile interpretation....

  11. Restricted Mobilities

    DEFF Research Database (Denmark)

    Nielsen, Mette; Lassen, Claus

    2012-01-01

    communities and shopping centres through mobility lenses. The article shows how different mobility systems enable and restrict the public access to private-public spaces, and it points out that proprietary communities create an unequal potential for human movement and access in the city. The main argument......Privatisation of public spaces in the contemporary city has increased during the last decades but only few studies have approached this field from a mobility perspective. Therefore the article seeks to rectify this by exploring two Australian examples of private spaces in the city; gated...... in the article is that the many mobility systems enable specialization of places that are targeted at a special section of the population. This means that various forms of motilities not only create new opportunities for urban life but it is also one of the most critical components of production of new exclusion...

  12. Staying Mobile

    Science.gov (United States)

    ... favorite. People who use scooters and wheelchairs bowl, fish, ski, and play golf, tennis or basketball. Enable ... the Independent Care System of NY addresses the importance of selecting the wheeled mobility device that best ...

  13. Mobile museology

    DEFF Research Database (Denmark)

    Baggesen, Rikke Haller

    Drawing together perspectives from museology, digital culture studies and fashion theory, this thesis considers changes in and challenges for current - day museums as related to ‘mobile museology’. This concept is developed for and elucidated in the thesis to describe an orientation towards...... the fashionable, the ephemeral, and towards an (ideal) state of change and changeability. This orientation is characterised with the triplet concepts of mobile, mobility, and mobilisation, as related to mobile media and movability; to ‘trans - museal’ mediation; and to the mobilisation of collections, audiences...... and institutional mindsets. The research project’s transdisciplinary and exploratory approach takes inspiration from critical design, minding Latour’s (2004a) call for rethink ing critical approaches in the humanities. Through a creative process, focused on designs for framing fashion in everyday contexts...

  14. Mobile Commerce

    Directory of Open Access Journals (Sweden)

    Maria Cristina Enache

    2016-07-01

    Full Text Available Mobile commerce, or m-commerce, refers to the use of wireless digital devices to enable transactions on the Web. Described more fully in Chapter 3, m-commerce involves the use of wireless networks to connect cell phones, handheld devices such Blackberries, and personal computers to the Web. Once connected, mobile consumers can conduct transactions, including stock trades, in-store price comparisons, banking, travel reservations, and more.

  15. Nucleosome assembly factors CAF-1 and HIR modulate epigenetic switching frequencies in an H3K56 acetylation-associated manner in Candida albicans.

    Science.gov (United States)

    Stevenson, John S; Liu, Haoping

    2013-04-01

    CAF-1 and HIR are highly conserved histone chaperone protein complexes that function in the assembly of nucleosomes onto chromatin. CAF-1 is characterized as having replication-coupled nucleosome activity, whereas the HIR complex can assemble nucleosomes independent of replication. Histone H3K56 acetylation, controlled by the acetyltransferase Rtt109 and deacetylase Hst3, also plays a significant role in nucleosome assembly. In this study, we generated a set of deletion mutants to genetically characterize pathway-specific and overlapping functions of CAF-1 and HIR in C. albicans. Their roles in epigenetic maintenance of cell type were examined by using the white-opaque switching system in C. albicans. We show that CAF-1 and HIR play conserved roles in UV radiation recovery, repression of histone gene expression, correct chromosome segregation, and stress responses. Unique to C. albicans, the cac2Δ/Δ mutant shows increased sensitivity to the Hst3 inhibitor nicotinamide, while the rtt109Δ/Δ cac2Δ/Δ and hir1Δ/Δ cac2Δ/Δ mutants are resistant to nicotinamide. CAF-1 plays a major role in maintaining cell types, as the cac2Δ/Δ mutant exhibited increased switching frequencies in both directions and switched at a high frequency to opaque in response to nicotinamide. Like the rtt109Δ/Δ mutant, the hir1Δ/Δ cac2Δ/Δ double mutant is defective in maintaining the opaque cell fate and blocks nicotinamide-induced opaque formation, and the defects are suppressed by ectopic expression of the master white-opaque regulator Wor1. Our data suggest an overlapping function of CAF-1 and HIR in epigenetic regulation of cell fate determination in an H3K56 acetylation-associated manner.

  16. The Ddc1-Mec3-Rad17 sliding clamp regulates histone-histone chaperone interactions and DNA replication-coupled nucleosome assembly in budding yeast.

    Science.gov (United States)

    Burgess, Rebecca J; Han, Junhong; Zhang, Zhiguo

    2014-04-11

    The maintenance of genome integrity is regulated in part by chromatin structure and factors involved in the DNA damage response pathway. Nucleosome assembly is a highly regulated process that restores chromatin structure after DNA replication, DNA repair, and gene transcription. During S phase the histone chaperones Asf1, CAF-1, and Rtt106 coordinate to deposit newly synthesized histones H3-H4 onto replicated DNA in budding yeast. Here we describe synthetic genetic interactions between RTT106 and the DDC1-MEC3-RAD17 (9-1-1) complex, a sliding clamp functioning in the S phase DNA damage and replication checkpoint response, upon treatment with DNA damaging agents. The DNA damage sensitivity of rad17Δ rtt106Δ cells depends on the function of Rtt106 in nucleosome assembly. Epistasis analysis reveals that 9-1-1 complex components interact with multiple DNA replication-coupled nucleosome assembly factors, including Rtt106, CAF-1, and lysine residues of H3-H4. Furthermore, rad17Δ cells exhibit defects in the deposition of newly synthesized H3-H4 onto replicated DNA. Finally, deletion of RAD17 results in increased association of Asf1 with checkpoint kinase Rad53, which may lead to the observed reduction in Asf1-H3 interaction in rad17Δ mutant cells. In addition, we observed that the interaction between histone H3-H4 with histone chaperone CAF-1 or Rtt106 increases in cells lacking Rad17. These results support the idea that the 9-1-1 checkpoint protein regulates DNA replication-coupled nucleosome assembly in part through regulating histone-histone chaperone interactions.

  17. Human OGG1 activity in nucleosomes is facilitated by transient unwrapping of DNA and is influenced by the local histone environment.

    Science.gov (United States)

    Bilotti, Katharina; Kennedy, Erin E; Li, Chuxuan; Delaney, Sarah

    2017-11-01

    If unrepaired, damage to genomic DNA can cause mutations and/or be cytotoxic. Single base lesions are repaired via the base excision repair (BER) pathway. The first step in BER is the recognition and removal of the nucleobase lesion by a glycosylase enzyme. For example, human oxoguanine glycosylase 1 (hOGG1) is responsible for removal of the prototypic oxidatively damaged nucleobase, 8-oxo-7,8-dihydroguanine (8-oxoG). To date, most studies of glycosylases have used free duplex DNA substrates. However, cellular DNA is packaged as repeating nucleosome units, with 145 base pair segments of DNA wrapped around histone protein octamers. Previous studies revealed inhibition of hOGG1 at the nucleosome dyad axis and in the absence of chromatin remodelers. In this study, we reveal that even in the absence of chromatin remodelers or external cofactors, hOGG1 can initiate BER at positions off the dyad axis and that this activity is facilitated by spontaneous and transient unwrapping of DNA from the histones. Additionally, we find that solution accessibility as determined by hydroxyl radical footprinting is not fully predictive of glycosylase activity and that histone tails can suppress hOGG1 activity. We therefore suggest that local nuances in the nucleosome environment and histone-DNA interactions can impact glycosylase activity. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Clipping of Flexible Tails of Histones H3 and H4 Affects the Structure and Dynamics of the Nucleosome

    Science.gov (United States)

    Nurse, Nathan P.; Jimenez-Useche, Isabel; Smith, Ian Tad; Yuan, Chongli

    2013-01-01

    Förster resonance energy transfer was used to monitor the dynamic conformations of mononucleosomes under different chromatin folding conditions to elucidate the role of the flexible N-terminal regions of H3 and H4 histones. The H3 tail was shown to partake in intranucleosomal interactions by restricting the DNA breathing motion and compacting the nucleosome. The H3 tail effects were mostly independent of the ionic strength and valency of the ions. The H4 tail was shown to not greatly affect the nucleosome conformation, but did slightly influence the relative population of the preferred conformation. The role of the H4 tail varied depending on the valency and ionic strength, suggesting that electrostatic forces play a primary role in H4 tail interactions. Interestingly, despite the H4 tail’s lack of influence, when H3 and H4 tails were simultaneously clipped, a more dramatic effect was seen than when only H3 or H4 tails were clipped. The combinatorial effect of H3 and H4 tail truncation suggests a potential mechanism by which various combinations of histone tail modifications can be used to control accessibility of DNA-binding proteins to nucleosomal DNA. PMID:23473491

  19. Lysine methyltransferase G9a is not required for DNMT3A/3B anchoring to methylated nucleosomes and maintenance of DNA methylation in somatic cells

    Directory of Open Access Journals (Sweden)

    Sharma Shikhar

    2012-01-01

    Full Text Available Abstract Background DNA methylation, histone modifications and nucleosome occupancy act in concert for regulation of gene expression patterns in mammalian cells. Recently, G9a, a H3K9 methyltransferase, has been shown to play a role in establishment of DNA methylation at embryonic gene targets in ES cells through recruitment of de novo DNMT3A/3B enzymes. However, whether G9a plays a similar role in maintenance of DNA methylation in somatic cells is still unclear. Results Here we show that G9a is not essential for maintenance of DNA methylation in somatic cells. Knockdown of G9a has no measurable effect on DNA methylation levels at G9a-target loci. DNMT3A/3B remain stably anchored to nucleosomes containing methylated DNA even in the absence of G9a, ensuring faithful propagation of methylated states in cooperation with DNMT1 through somatic divisions. Moreover, G9a also associates with nucleosomes in a DNMT3A/3B and DNA methylation-independent manner. However, G9a knockdown synergizes with pharmacologic inhibition of DNMTs resulting in increased hypomethylation and inhibition of cell proliferation. Conclusions Taken together, these data suggest that G9a is not involved in maintenance of DNA methylation in somatic cells but might play a role in re-initiation of de novo methylation after treatment with hypomethylating drugs, thus serving as a potential target for combinatorial treatments strategies involving DNMTs inhibitors.

  20. Major satellite repeat RNA stabilize heterochromatin retention of Suv39h enzymes by RNA-nucleosome association and RNA:DNA hybrid formation.

    Science.gov (United States)

    Velazquez Camacho, Oscar; Galan, Carmen; Swist-Rosowska, Kalina; Ching, Reagan; Gamalinda, Michael; Karabiber, Fethullah; De La Rosa-Velazquez, Inti; Engist, Bettina; Koschorz, Birgit; Shukeir, Nicholas; Onishi-Seebacher, Megumi; van de Nobelen, Suzanne; Jenuwein, Thomas

    2017-08-01

    The Suv39h1 and Suv39h2 histone lysine methyltransferases are hallmark enzymes at mammalian heterochromatin. We show here that the mouse Suv39h2 enzyme differs from Suv39h1 by containing an N-terminal basic domain that facilitates retention at mitotic chromatin and provides an additional affinity for major satellite repeat RNA. To analyze an RNA-dependent interaction with chromatin, we purified native nucleosomes from mouse ES cells and detect that Suv39h1 and Suv39h2 exclusively associate with poly-nucleosomes. This association was attenuated upon RNaseH incubation and entirely lost upon RNaseA digestion of native chromatin. Major satellite repeat transcripts remain chromatin-associated and have a secondary structure that favors RNA:DNA hybrid formation. Together, these data reveal an RNA-mediated mechanism for the stable chromatin interaction of the Suv39h KMT and suggest a function for major satellite non-coding RNA in the organization of an RNA-nucleosome scaffold as the underlying structure of mouse heterochromatin.

  1. Mobile healthcare.

    Science.gov (United States)

    Morgan, Stephen A; Agee, Nancy Howell

    2012-01-01

    Mobile technology's presence in healthcare has exploded over the past five years. The increased use of mobile devices by all segments of the US population has driven healthcare systems, providers, and payers to accept this new form of communication and to develop strategies to implement and leverage the use of mobile healthcare (mHealth) within their organizations and practices. As healthcare systems move toward a more value-driven model of care, patient centeredness and engagement are the keys to success. Mobile healthcare will provide the medium to allow patients to participate more in their care. Financially, mHealth brings to providers the ability to improve efficiency and deliver savings to both them and the healthcare consumer. However, mHealth is not without challenges. Healthcare IT departments have been reluctant to embrace this shift in technology without fully addressing security and privacy concerns. Providers have been hesitant to adopt mHealth as a form of communication with patients because it breaks with traditional models. Our healthcare system has just started the journey toward the development of mHealth. We offer an overview of the mobile healthcare environment and our approach to solving the challenges it brings to healthcare organizations.

  2. BAF53b, a Neuron-Specific Nucleosome Remodeling Factor, Is Induced after Learning and Facilitates Long-Term Memory Consolidation.

    Science.gov (United States)

    Yoo, Miran; Choi, Kwang-Yeon; Kim, Jieun; Kim, Mujun; Shim, Jaehoon; Choi, Jun-Hyeok; Cho, Hye-Yeon; Oh, Jung-Pyo; Kim, Hyung-Su; Kaang, Bong-Kiun; Han, Jin-Hee

    2017-03-29

    Although epigenetic mechanisms of gene expression regulation have recently been implicated in memory consolidation and persistence, the role of nucleosome-remodeling is largely unexplored. Recent studies show that the functional loss of BAF53b, a postmitotic neuron-specific subunit of the BAF nucleosome-remodeling complex, results in the deficit of consolidation of hippocampus-dependent memory and cocaine-associated memory in the rodent brain. However, it is unclear whether BAF53b expression is regulated during memory formation and how BAF53b regulates fear memory in the amygdala, a key brain site for fear memory encoding and storage. To address these questions, we used viral vector approaches to either decrease or increase BAF53b function specifically in the lateral amygdala of adult mice in auditory fear conditioning paradigm. Knockdown of Baf53b before training disrupted long-term memory formation with no effect on short-term memory, basal synaptic transmission, and spine structures. We observed in our qPCR analysis that BAF53b was induced in the lateral amygdala neurons at the late consolidation phase after fear conditioning. Moreover, transient BAF53b overexpression led to persistently enhanced memory formation, which was accompanied by increase in thin-type spine density. Together, our results provide the evidence that BAF53b is induced after learning, and show that such increase of BAF53b level facilitates memory consolidation likely by regulating learning-related spine structural plasticity.SIGNIFICANCE STATEMENT Recent works in the rodent brain begin to link nucleosome remodeling-dependent epigenetic mechanism to memory consolidation. Here we show that BAF53b, an epigenetic factor involved in nucleosome remodeling, is induced in the lateral amygdala neurons at the late phase of consolidation after fear conditioning. Using specific gene knockdown or overexpression approaches, we identify the critical role of BAF53b in the lateral amygdala neurons for memory

  3. Designing Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    are often still not engaged with in a sufficiently manner. Often social sciences keep distance to the physical and material as if the social was still to be understood as a realm separate of technology, architecture, and design (for a critique of this see; Latour 2005 and Urry 2000). This paper takes point...... of departure in the sociological perspective termed ‘Staging Mobilities’ (Jensen 2013a) and utilizes this as an analytical frame for exploring cases of mobility design. The paper put focus on how the material shape, design and architectures of technologies, spaces and sites influence mobilities practices......, mobilitiy technologies or urban sites of movement we get much closer to understanding the meaning of mobilities to social interaction and culture. The cases are still representing work-in-progress but will be reported in the book ‘Designing Mobilites’ (Jensen 2013b) and will cover the four cases of...

  4. The NuRD nucleosome remodelling complex and NHK-1 kinase are required for chromosome condensation in oocytes.

    Science.gov (United States)

    Nikalayevich, Elvira; Ohkura, Hiroyuki

    2015-02-01

    Chromosome condensation during cell division is one of the most dramatic events in the cell cycle. Condensin and topoisomerase II are the most studied factors in chromosome condensation. However, their inactivation leads to only mild defects and little is known about the roles of other factors. Here, we took advantage of Drosophilaoocytes to elucidate the roles of potential condensation factors by performing RNA interference (RNAi). Consistent with previous studies, depletion of condensin I subunits or topoisomerase II in oocytes only mildly affected chromosome condensation. In contrast, we found severe undercondensation of chromosomes after depletion of the Mi-2-containing NuRD nucleosome remodelling complex or the protein kinase NHK-1 (also known as Ballchen in Drosophila). The further phenotypic analysis suggests that Mi-2 and NHK-1 are involved in different pathways of chromosome condensation. We show that the main role of NHK-1 in chromosome condensation is to phosphorylate Barrier-to-autointegration factor (BAF) and suppress its activity in linking chromosomes to nuclear envelope proteins. We further show that NHK-1 is important for chromosome condensation during mitosis as well as in oocytes.

  5. A DNA Structural Alphabet Distinguishes Structural Features of DNA Bound to Regulatory Proteins and in the Nucleosome Core Particle.

    Science.gov (United States)

    Schneider, Bohdan; Božíková, Paulína; Čech, Petr; Svozil, Daniel; Černý, Jiří

    2017-10-18

    We analyzed the structural behavior of DNA complexed with regulatory proteins and the nucleosome core particle (NCP). The three-dimensional structures of almost 25 thousand dinucleotide steps from more than 500 sequentially non-redundant crystal structures were classified by using DNA structural alphabet CANA (Conformational Alphabet of Nucleic Acids) and associations between ten CANA letters and sixteen dinucleotide sequences were investigated. The associations showed features discriminating between specific and non-specific binding of DNA to proteins. Important is the specific role of two DNA structural forms, A-DNA, and BII-DNA, represented by the CANA letters AAA and BB2: AAA structures are avoided in non-specific NCP complexes, where the wrapping of the DNA duplex is explained by the periodic occurrence of BB2 every 10.3 steps. In both regulatory and NCP complexes, the extent of bending of the DNA local helical axis does not influence proportional representation of the CANA alphabet letters, namely the relative incidences of AAA and BB2 remain constant in bent and straight duplexes.

  6. Splice variants of the SWR1-type nucleosome remodeling factor Domino have distinct functions during Drosophila melanogaster oogenesis.

    Science.gov (United States)

    Börner, Kenneth; Becker, Peter B

    2016-09-01

    SWR1-type nucleosome remodeling factors replace histone H2A by variants to endow chromatin locally with specialized functionality. In Drosophila melanogaster a single H2A variant, H2A.V, combines functions of mammalian H2A.Z and H2A.X in transcription regulation and the DNA damage response. A major role in H2A.V incorporation for the only SWR1-like enzyme in flies, Domino, is assumed but not well documented in vivo. It is also unclear whether the two alternatively spliced isoforms, DOM-A and DOM-B, have redundant or specialized functions. Loss of both DOM isoforms compromises oogenesis, causing female sterility. We systematically explored roles of the two DOM isoforms during oogenesis using a cell type-specific knockdown approach. Despite their ubiquitous expression, DOM-A and DOM-B have non-redundant functions in germline and soma for egg formation. We show that chromatin incorporation of H2A.V in germline and somatic cells depends on DOM-B, whereas global incorporation in endoreplicating germline nurse cells appears to be independent of DOM. By contrast, DOM-A promotes the removal of H2A.V from stage 5 nurse cells. Remarkably, therefore, the two DOM isoforms have distinct functions in cell type-specific development and H2A.V exchange. © 2016. Published by The Company of Biologists Ltd.

  7. The Nucleosome Remodeling and Deacetylase Complex NuRD Is Built from Preformed Catalytically Active Sub-modules.

    Science.gov (United States)

    Zhang, W; Aubert, A; Gomez de Segura, J M; Karuppasamy, M; Basu, S; Murthy, A S; Diamante, A; Drury, T A; Balmer, J; Cramard, J; Watson, A A; Lando, D; Lee, S F; Palayret, M; Kloet, S L; Smits, A H; Deery, M J; Vermeulen, M; Hendrich, B; Klenerman, D; Schaffitzel, C; Berger, I; Laue, E D

    2016-07-17

    The nucleosome remodeling deacetylase (NuRD) complex is a highly conserved regulator of chromatin structure and transcription. Structural studies have shed light on this and other chromatin modifying machines, but much less is known about how they assemble and whether stable and functional sub-modules exist that retain enzymatic activity. Purification of the endogenous Drosophila NuRD complex shows that it consists of a stable core of subunits, while others, in particular the chromatin remodeler CHD4, associate transiently. To dissect the assembly and activity of NuRD, we systematically produced all possible combinations of different components using the MultiBac system, and determined their activity and biophysical properties. We carried out single-molecule imaging of CHD4 in live mouse embryonic stem cells, in the presence and absence of one of core components (MBD3), to show how the core deacetylase and chromatin-remodeling sub-modules associate in vivo. Our experiments suggest a pathway for the assembly of NuRD via preformed and active sub-modules. These retain enzymatic activity and are present in both the nucleus and the cytosol, an outcome with important implications for understanding NuRD function. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  8. Sall4 controls differentiation of pluripotent cells independently of the Nucleosome Remodelling and Deacetylation (NuRD) complex.

    Science.gov (United States)

    Miller, Anzy; Ralser, Meryem; Kloet, Susan L; Loos, Remco; Nishinakamura, Ryuichi; Bertone, Paul; Vermeulen, Michiel; Hendrich, Brian

    2016-09-01

    Sall4 is an essential transcription factor for early mammalian development and is frequently overexpressed in cancer. Although it is reported to play an important role in embryonic stem cell (ESC) self-renewal, whether it is an essential pluripotency factor has been disputed. Here, we show that Sall4 is dispensable for mouse ESC pluripotency. Sall4 is an enhancer-binding protein that prevents precocious activation of the neural gene expression programme in ESCs but is not required for maintenance of the pluripotency gene regulatory network. Although a proportion of Sall4 protein physically associates with the Nucleosome Remodelling and Deacetylase (NuRD) complex, Sall4 neither recruits NuRD to chromatin nor influences transcription via NuRD; rather, free Sall4 protein regulates transcription independently of NuRD. We propose a model whereby enhancer binding by Sall4 and other pluripotency-associated transcription factors is responsible for maintaining the balance between transcriptional programmes in pluripotent cells. © 2016. Published by The Company of Biologists Ltd.

  9. Inferring coarse-grain histone-DNA interaction potentials from high-resolution structures of the nucleosome

    CERN Document Server

    Meyer, Sam

    2014-01-01

    The histone-DNA interaction in the nucleosome is a fundamental mechanism of genomic compaction and regulation, which remains largely unkown despite a growing structural knowledge of the complex. Here, we propose a framework for the extraction of a nanoscale histone-DNA force-field from a collection of high-resolution structures, which may be adapted to a larger class of protein-DNA complexes. We apply the procedure on a large crystallographic database extended by snapshots from molecular dynamics simulations. The comparison of the structural models first shows that, at the sites of histone-DNA contact, the DNA base-pairs are locally shifted outwards, consistent with locally repulsive forces exerted by the histones. In a second step, we show that the various force profiles of the analyzed structures derive locally from a unique, sequence-independent, quadratic repulsive force field, while the sequence preferences are entirely due to the internal DNA mechanics. We thus obtain the first knowledge-derived nanosca...

  10. Structural Architecture of the Nucleosome Remodeler ISWI Determined from Cross-Linking, Mass Spectrometry, SAXS, and Modeling.

    Science.gov (United States)

    Harrer, Nadine; Schindler, Christina E M; Bruetzel, Linda K; Forné, Ignasi; Ludwigsen, Johanna; Imhof, Axel; Zacharias, Martin; Lipfert, Jan; Mueller-Planitz, Felix

    2018-02-06

    Chromatin remodeling factors assume critical roles by regulating access to nucleosomal DNA. To determine the architecture of the Drosophila ISWI remodeling enzyme, we developed an integrative structural approach that combines protein cross-linking, mass spectrometry, small-angle X-ray scattering, and computational modeling. The resulting structural model shows the ATPase module in a resting state with both ATPase lobes twisted against each other, providing support for a conformation that was recently trapped by crystallography. The autoinhibiting NegC region does not protrude from the ATPase module as suggested previously. The regulatory NTR domain is located near both ATPase lobes. The full-length enzyme is flexible and can adopt a compact structure in solution with the C-terminal HSS domain packing against the ATPase module. Our data imply a series of conformational changes upon activation of the enzyme and illustrate how the NTR, NegC, and HSS domains contribute to regulation of the ATPase module. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Transcription factor 19 interacts with histone 3 lysine 4 trimethylation and controls gluconeogenesis via the nucleosome-remodeling-deacetylase complex.

    Science.gov (United States)

    Sen, Sabyasachi; Sanyal, Sulagna; Srivastava, Dushyant Kumar; Dasgupta, Dipak; Roy, Siddhartha; Das, Chandrima

    2017-12-15

    Transcription factor 19 (TCF19) has been reported as a type 1 diabetes-associated locus involved in maintenance of pancreatic β cells through a fine-tuned regulation of cell proliferation and apoptosis. TCF19 also exhibits genomic association with type 2 diabetes, although the precise molecular mechanism remains unknown. It harbors both a plant homeodomain and a forkhead-associated domain implicated in epigenetic recognition and gene regulation, a phenomenon that has remained unexplored. Here, we show that TCF19 selectively interacts with histone 3 lysine 4 trimethylation through its plant homeodomain finger. Knocking down TCF19 under high-glucose conditions affected many metabolic processes, including gluconeogenesis. We found that TCF19 overexpression represses de novo glucose production in HepG2 cells. The transcriptional repression of key genes, induced by TCF19, coincided with NuRD (nucleosome-remodeling-deacetylase) complex recruitment to the promoters of these genes. TCF19 interacted with CHD4 (chromodomain helicase DNA-binding protein 4), which is a part of the NuRD complex, in a glucose concentration-independent manner. In summary, our results show that TCF19 interacts with an active transcription mark and recruits a co-repressor complex to regulate gluconeogenic gene expression in HepG2 cells. Our study offers critical insights into the molecular mechanisms of transcriptional regulation of gluconeogenesis and into the roles of chromatin readers in metabolic homeostasis. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  12. Analysis of a Polycomb Group Protein Defines Regions That Link Repressive Activity on Nucleosomal Templates to In Vivo Function

    Science.gov (United States)

    King, Ian F. G.; Emmons, Richard B.; Francis, Nicole J.; Wild, Brigitte; Müller, Jürg; Kingston, Robert E.; Wu, Chao-ting

    2005-01-01

    Polycomb group (PcG) genes propagate patterns of transcriptional repression throughout development. The products of several such genes are part of Polycomb repressive complex 1 (PRC1), which inhibits chromatin remodeling and transcription in vitro. Genetic and biochemical studies suggest the product of the Posterior sex combs (Psc) gene plays a central role in both PcG-mediated gene repression in vivo and PRC1 activity in vitro. To dissect the relationship between the in vivo and in vitro activities of Psc, we identified the lesions associated with 11 genetically characterized Psc mutations and asked how the corresponding mutant proteins affect Psc activity on nucleosomal templates in vitro. Analysis of both single-mutant Psc proteins and recombinant complexes containing mutant protein revealed that Psc encodes at least two functions, complex formation and the inhibition of remodeling and transcription, which require different regions of the protein. There is an excellent correlation between the in vivo phenotypes of mutant Psc alleles and the structure and in vitro activities of the corresponding proteins, suggesting that the in vitro activities of PRC1 reflect essential functions of Psc in vivo. PMID:16024794

  13. Histone H3K56 acetylation, CAF1, and Rtt106 coordinate nucleosome assembly and stability of advancing replication forks.

    Directory of Open Access Journals (Sweden)

    Marta Clemente-Ruiz

    2011-11-01

    Full Text Available Chromatin assembly mutants accumulate recombinogenic DNA damage and are sensitive to genotoxic agents. Here we have analyzed why impairment of the H3K56 acetylation-dependent CAF1 and Rtt106 chromatin assembly pathways, which have redundant roles in H3/H4 deposition during DNA replication, leads to genetic instability. We show that the absence of H3K56 acetylation or the simultaneous knock out of CAF1 and Rtt106 increases homologous recombination by affecting the integrity of advancing replication forks, while they have a minor effect on stalled replication fork stability in response to the replication inhibitor hydroxyurea. This defect in replication fork integrity is not due to defective checkpoints. In contrast, H3K56 acetylation protects against replicative DNA damaging agents by DNA repair/tolerance mechanisms that do not require CAF1/Rtt106 and are likely subsequent to the process of replication-coupled nucleosome deposition. We propose that the tight connection between DNA synthesis and histone deposition during DNA replication mediated by H3K56ac/CAF1/Rtt106 provides a mechanism for the stabilization of advancing replication forks and the maintenance of genome integrity, while H3K56 acetylation has an additional, CAF1/Rtt106-independent function in the response to replicative DNA damage.

  14. Resistance to therapy in BRCA2 mutant cells due to loss of the nucleosome remodeling factor CHD4.

    Science.gov (United States)

    Guillemette, Shawna; Serra, Ryan W; Peng, Min; Hayes, Janelle A; Konstantinopoulos, Panagiotis A; Green, Michael R; Cantor, Sharon B

    2015-03-01

    Hereditary cancers derive from gene defects that often compromise DNA repair. Thus, BRCA-associated cancers are sensitive to DNA-damaging agents such as cisplatin. The efficacy of cisplatin is limited, however, by the development of resistance. One cisplatin resistance mechanism is restoration of homologous recombination (HR), which can result from BRCA reversion mutations. However, in BRCA2 mutant cancers, cisplatin resistance can occur independently of restored HR by a mechanism that remains unknown. Here we performed a genome-wide shRNA screen and found that loss of the nucleosome remodeling factor CHD4 confers cisplatin resistance. Restoration of cisplatin resistance is independent of HR but correlates with restored cell cycle progression, reduced chromosomal aberrations, and enhanced DNA damage tolerance. Suggesting clinical relevance, cisplatin-resistant clones lacking genetic reversion of BRCA2 show de novo loss of CHD4 expression in vitro. Moreover, BRCA2 mutant ovarian cancers with reduced CHD4 expression significantly correlate with shorter progression-free survival and shorter overall survival. Collectively, our findings indicate that CHD4 modulates therapeutic response in BRCA2 mutant cancer cells. © 2015 Guillemette et al.; Published by Cold Spring Harbor Laboratory Press.

  15. Mobile probes

    DEFF Research Database (Denmark)

    Ørngreen, Rikke; Jørgensen, Anna Neustrup; Noesgaard, Signe Schack

    2016-01-01

    to in an interview. This method provided valuable insight into the contextual use, i.e. how did the online resource transfer to the work practice. However, the research team also found that mobile probes may provide the scaffolding necessary for individual and peer learning at a very local (intra-school) community...

  16. Designing Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    How is the width of the pavement shaping the urban experience? How is the material design of transport infrastructure and mobile technology affording social interaction in everyday life spaces? How do people inhabit these spaces with their bodies and in accordance to social and cultural norms...

  17. Mobile Misfortune

    DEFF Research Database (Denmark)

    Vigh, Henrik Erdman

    2015-01-01

    of the mobility it enables. This article, thus, looks at the motives and manners in which young men in Bissau become caught up in transnational flows of cocaine. It shows how motion is emotively anchored and affectively bound: tied to and directed toward a feeling of worth and realisation of being, and how...

  18. Mobile IP

    NARCIS (Netherlands)

    Heijenk, Geert; Sallent, S.; Pras, Aiko

    1999-01-01

    The Internet is growing exponentially, both in the amount of traffic carried, and in the amount of hosts connected. IP technology is becoming more and more important, in company networks (Intranets), and also in the core networks for the next generation mobile networks. Further, wireless access to

  19. Mobile-to-mobile wireless channels

    CERN Document Server

    Zajic, Alenka

    2013-01-01

    Present-day mobile communications systems can be classified as fixed-to-mobile because they allow mobility on only one end (e.g. the mobile phone to a fixed mobile operator's cell tower). In answer to the consumer demand for better coverage and quality of service, emerging mobile-to-mobile (M-to-M) communications systems allow mobile users or vehicles to directly communicate with each other. This practical book provides a detailed introduction to state-of-the-art M-to-M wireless propagation. Moreover, the book offers professionals guidance for rapid implementation of these communications syste

  20. Mobile Customer Relationship Management and Mobile Security

    Science.gov (United States)

    Sanayei, Ali; Mirzaei, Abas

    The purpose of this study is twofold. First, in order to guarantee a coherent discussion about mobile customer relationship management (mCRM), this paper presents a conceptualization of mCRM delineating its unique characteristics because of Among the variety of mobile services, considerable attention has been devoted to mobile marketing and in particular to mobile customer relationship management services. Second, the authors discusses the security risks in mobile computing in different level(user, mobile device, wireless network,...) and finally we focus on enterprise mobile security and it's subgroups with a series of suggestion and solution for improve mobile computing security.

  1. The African Mobile Story

    DEFF Research Database (Denmark)

    This book identifies the factors that has enabled the growth of mobile telephony in Africa. The book covers the regulatory factors, the development and usage of mobile application, mobile security and sustainable power source for mobile networks......This book identifies the factors that has enabled the growth of mobile telephony in Africa. The book covers the regulatory factors, the development and usage of mobile application, mobile security and sustainable power source for mobile networks...

  2. Urban Mobility

    DEFF Research Database (Denmark)

    2017-01-01

    This anthology is the proceedings publication from the 2015 NAF Symposium in Malmö, Sweden. The aim of the 2015 NAF Symposium “Urban Mobility – Architectures, Geographies and Social Space” was to facilitate a cross-disciplinary discussion on urban mobility in which the juxtaposition of different...... discursive perceptions of the concept would foster greater insight into and understanding of both the challenges and potentials that it represents. It focused on some of the key themes currently facing cities and the urban: the transformation of the city and our built environment; migration; rural decline......; the interaction between city, architecture, and inhabitants; the role of architects and architecture in the creation of democratic and sustainable urban contexts; the city and its representation; the politics of intervention; and the actions of governing and developing. This proceedings publication from...

  3. Structural Mobility, Exchange Mobility and Subgroup Consistent Mobility Measurement – US–German Mobility Measurements Revisited

    OpenAIRE

    Schluter, C.; D. VAN DE GAER

    2008-01-01

    We formalize the concept of structural mobility and use the framework of subgroup consistent mobility measurement to derive a relative and an absolute measure of mobility that is increasing both in upward structural mobility and exchange mobility. In our empirical illustration, we contribute substantively to the ongoing debate about mobility rankings between the USA and Germany.

  4. Vaccination with a plasmid DNA cocktail encoding the nucleosomal histones of Leishmania confers protection against murine cutaneous leishmaniosis.

    Science.gov (United States)

    Iborra, Salvador; Soto, Manuel; Carrión, Javier; Alonso, Carlos; Requena, Jose M

    2004-09-28

    Leishmania histones are relevant immunogens for the host immune system during both Leishmania infection and disease. In the present paper we have evaluated the prophylactic value of the four Leishmania infantum histones forming the nucleosomal core in the murine model of cutaneous leishmaniasis. In a first stage, the immune response elicited by the intramuscular injection of a mixture of four plasmid DNAs, encoding the L. infantum histones H2A, H2B, H3 and H4, was determined in BALB/c mice. It was found that the immunized animals developed a specific Th1 immune response, which was associated with an antigen-specific production of interferon (IFN-gamma) and a limited humoral response against histones (dominated by antibodies of the IgG2a isotype). According to the pure Th1-type immune response elicited by the DNA vaccination with Leishmania histones, vaccinated mice showed a solid immunity that efficiently controlled the Leishmania major infection. The protection in mice vaccinated with histone-DNAs was associated with a low humoral response against leishmanial antigens, an enhanced IFN-gamma production and little, if any, IL-4 production. The relative contribution of both CD8(+) and CD4(+) T cells to the IFN-gamma production, and the IL-12 dependence were also evaluated. All these data indicated that DNA vaccination with Leishmania histones genes results in a specific Th1-like response during L. major infection, and that both CD4(+) and CD8(+) T cells contribute to the resistance of vaccinated mice to cutaneous leishmaniasis.

  5. Changes in nucleosome repeat lengths precede replication in the early replicating metallothionein II gene region of cells synchronized in early S phase

    Energy Technology Data Exchange (ETDEWEB)

    D' Anna, J.A.; Tobey, R.A. (Los Alamos National Lab., NM (USA))

    1989-04-04

    Previous investigations showed that inhibition of DNA synthesis by hydroxyurea, aphidicolin, or 5-fluorodeoxyuridine produced large changes in the composition and nucleosome repeat lengths of bulk chromatin. There the authors report results of investigations to determine whether the changes in nucleosome repeat lengths might be localized in the initiated replicons, as postulated. In most experiments, Chinese hamster (line CHO) cells were synchronized in G1, or they were synchronized in early S phase by allowing G1 cells to enter S phase in medium containing 1 mM hydroxyurea or 5 {mu}g mL{sup {minus}1} aphidicolin, a procedure believed to produce an accumulation of initiated replicons that arise from normally early replicating DNA. Measurements of nucleosome repeat lengths of bulk chromatin, the early replicating unexpressed metallothionein II (MTII) gene region, and a later replicating repeated sequence indicate that the changes in repeat lengths occur preferentially in the early replicating MTII gene region as G1 cells enter and become synchronized in early S phase. During that time, the MTII gene region is not replicated nor is there any evidence for induction of MTII messenger RNA. Thus, the results are consistent with the hypothesis that changes in chromatin structure occur preferentially in the early replicating (presumably initiated) replicons at initiation or that changes in chromatin structure can precede replication during inhibition of DNA synthesis. The shortened repeat lengths that precede MTII replication are, potentially, reversible, because they become elongated when the synchronized early S-phase cells are released to resume cell cycle progression.

  6. The male germ cell gene regulator CTCFL is functionally different from CTCF and binds CTCF-like consensus sites in a nucleosome composition-dependent manner.

    Science.gov (United States)

    Sleutels, Frank; Soochit, Widia; Bartkuhn, Marek; Heath, Helen; Dienstbach, Sven; Bergmaier, Philipp; Franke, Vedran; Rosa-Garrido, Manuel; van de Nobelen, Suzanne; Caesar, Lisa; van der Reijden, Michael; Bryne, Jan Christian; van Ijcken, Wilfred; Grootegoed, J Anton; Delgado, M Dolores; Lenhard, Boris; Renkawitz, Rainer; Grosveld, Frank; Galjart, Niels

    2012-06-18

    CTCF is a highly conserved and essential zinc finger protein expressed in virtually all cell types. In conjunction with cohesin, it organizes chromatin into loops, thereby regulating gene expression and epigenetic events. The function of CTCFL or BORIS, the testis-specific paralog of CTCF, is less clear. Using immunohistochemistry on testis sections and fluorescence-based microscopy on intact live seminiferous tubules, we show that CTCFL is only transiently present during spermatogenesis, prior to the onset of meiosis, when the protein co-localizes in nuclei with ubiquitously expressed CTCF. CTCFL distribution overlaps completely with that of Stra8, a retinoic acid-inducible protein essential for the propagation of meiosis. We find that absence of CTCFL in mice causes sub-fertility because of a partially penetrant testicular atrophy. CTCFL deficiency affects the expression of a number of testis-specific genes, including Gal3st1 and Prss50. Combined, these data indicate that CTCFL has a unique role in spermatogenesis. Genome-wide RNA expression studies in ES cells expressing a V5- and GFP-tagged form of CTCFL show that genes that are downregulated in CTCFL-deficient testis are upregulated in ES cells. These data indicate that CTCFL is a male germ cell gene regulator. Furthermore, genome-wide DNA-binding analysis shows that CTCFL binds a consensus sequence that is very similar to that of CTCF. However, only ~3,700 out of the ~5,700 CTCFL- and ~31,000 CTCF-binding sites overlap. CTCFL binds promoters with loosely assembled nucleosomes, whereas CTCF favors consensus sites surrounded by phased nucleosomes. Finally, an ES cell-based rescue assay shows that CTCFL is functionally different from CTCF. Our data suggest that nucleosome composition specifies the genome-wide binding of CTCFL and CTCF. We propose that the transient expression of CTCFL in spermatogonia and preleptotene spermatocytes serves to occupy a subset of promoters and maintain the expression of male germ

  7. The ATP-dependent chromatin remodeling enzymes CHD6, CHD7, and CHD8 exhibit distinct nucleosome binding and remodeling activities.

    Science.gov (United States)

    Manning, Benjamin J; Yusufzai, Timur

    2017-07-14

    Proper chromatin regulation is central to genome function and maintenance. The group III chromodomain-helicase-DNA-binding (CHD) family of ATP-dependent chromatin remodeling enzymes, comprising CHD6, CHD7, CHD8, and CHD9, has well-documented roles in transcription regulation, impacting both organism development and disease etiology. These four enzymes are similar in their constituent domains, but they fill surprisingly non-redundant roles in the cell, with deficiencies in individual enzymes leading to dissimilar disease states such as CHARGE syndrome or autism spectrum disorders. The mechanisms explaining their divergent, non-overlapping functions are unclear. In this study, we performed an in-depth biochemical analysis of purified CHD6, CHD7, and CHD8 and discovered distinct differences in chromatin remodeling specificities and activities among them. We report that CHD6 and CHD7 both bind with high affinity to short linker DNA, whereas CHD8 requires longer DNA for binding. As a result, CHD8 slides nucleosomes into positions with more flanking linker DNA than CHD7. Moreover, we found that, although CHD7 and CHD8 slide nucleosomes, CHD6 disrupts nucleosomes in a distinct non-sliding manner. The different activities of these enzymes likely lead to differences in chromatin structure and, thereby, transcriptional control, at the enhancer and promoter loci where these enzymes bind. Overall, our work provides a mechanistic basis for both the non-redundant roles and the diverse mutant disease states of these enzymes in vivo. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  8. Mobile termination and mobile penetration

    OpenAIRE

    Hurkens, Sjaak; Jeon, Doh-Shin

    2009-01-01

    In this paper, we study how access pricing affects network competition when subscription demand is elastic and each network uses non-linear prices and can apply termination-based price discrimination. In the case of a fixed per minute termination charge, we find that a reduction of the termination charge below cost has two oppos- ing effects: it softens competition but helps to internalize network externalities. The former reduces mobile penetration while the latter boosts it. We find that fi...

  9. Mobility Bibliography.

    Science.gov (United States)

    1981-11-01

    IV-419 Strauss, A.M. - V-55 Stuller, J. - IV-470 Stuller, J.G. - IV-457 Sugarman , R.C. - IV-707 Suhier, S.A. - IV-540 Sullivan, E. - 111-214 Sullivan...military vehicle mobility 148 110. AU - Brannon, W.; David , R.H.; Hodges, W., Jr.; Janowski, W.R. TI - Design and development of the twister testbed SO...OS - Aluminum Co of America Alcoa Center PA, David W. Taylor Naval Ship Research and Development Center, Bethesda, MD SO - 20 Feb 76, 59 p, ADA024262

  10. Downwardly mobile

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1999-01-01

    P & H Mining Equipment has produced the 250XP rotary blasthole drill, its first `clean sheet` drill after eight years of upgrading the line of large rotaries it acquired from Gardner Denver in 1991. A prototype has been working on an Australian coal mine since June 1998. A further two units of the 250XP are being erected at a diamond mine in Botswana for start-up in January 1999 and another will begin drilling at a Wyoming, USA coal mine in February. The drill is a highly mobile, heavy duty, highly reliable diesel/hydraulic drill weighing 113,500 kg and can drill holes at angles up to 30 degrees.

  11. Wapice News Mobile Application

    OpenAIRE

    Söylemez, Ilke

    2017-01-01

    Since the mobile phones started to have an increasingly significant role in daily life, the mobile application development also started to be an important area in the software industry. The problem for mobile application developers is to develop a mobile application which supports all the devices and platforms on the market. This issue created a need for cross platform mobile applications. The cross platform mobile development refers to the development of mobile applications that could be use...

  12. High Mobility Group A2 protects cancer cells against telomere dysfunction

    Science.gov (United States)

    Natarajan, Suchitra; Begum, Farhana; Gim, Jeonga; Wark, Landon; Henderson, Dana; Davie, James R.

    2016-01-01

    The non-histone chromatin binding protein High Mobility Group AT-hook protein 2 (HMGA2) plays important roles in the repair and protection of genomic DNA in embryonic stem cells and cancer cells. Here we show that HMGA2 localizes to mammalian telomeres and enhances telomere stability in cancer cells. We present a novel interaction of HMGA2 with the key shelterin protein TRF2. We found that the linker (L1) region of HMGA2 contributes to this interaction but the ATI-L1-ATII molecular region of HMGA2 is required for strong interaction with TRF2. This interaction was independent of HMGA2 DNA-binding and did not require the TRF2 interacting partner RAP1 but involved the homodimerization and hinge regions of TRF2. HMGA2 retained TRF2 at telomeres and reduced telomere-dysfunction despite induced telomere stress. Silencing of HMGA2 resulted in (i) reduced binding of TRF2 to telomere DNA as observed by ChIP, (ii) increased telomere instability and (iii) the formation of telomere dysfunction-induced foci (TIF). This resulted in increased telomere aggregation, anaphase bridges and micronuclei. HMGA2 prevented ATM-dependent pTRF2T188 phosphorylation and attenuated signaling via the telomere specific ATM-CHK2-CDC25C DNA damage signaling axis. In summary, our data demonstrate a unique and novel role of HMGA2 in telomere protection and promoting telomere stability in cancer cells. This identifies HMGA2 as a new therapeutic target for the destabilization of telomeres in HMGA2+ cancer cells. PMID:26799419

  13. Eaf5/7/3 form a functionally independent NuA4 submodule linked to RNA polymerase II-coupled nucleosome recycling.

    Science.gov (United States)

    Rossetto, Dorine; Cramet, Myriam; Wang, Alice Y; Steunou, Anne-Lise; Lacoste, Nicolas; Schulze, Julia M; Côté, Valérie; Monnet-Saksouk, Julie; Piquet, Sandra; Nourani, Amine; Kobor, Michael S; Côté, Jacques

    2014-06-17

    The NuA4 histone acetyltransferase complex is required for gene regulation, cell cycle progression, and DNA repair. Dissection of the 13-subunit complex reveals that the Eaf7 subunit bridges Eaf5 with Eaf3, a H3K36me3-binding chromodomain protein, and this Eaf5/7/3 trimer is anchored to NuA4 through Eaf5. This trimeric subcomplex represents a functional module, and a large portion exists in a native form outside the NuA4 complex. Gene-specific and genome-wide location analyses indicate that Eaf5/7/3 correlates with transcription activity and is enriched over the coding region. In agreement with a role in transcription elongation, the Eaf5/7/3 trimer interacts with phosphorylated RNA polymerase II and helps its progression. Loss of Eaf5/7/3 partially suppresses intragenic cryptic transcription arising in set2 mutants, supporting a role in nucleosome destabilization. On the other hand, loss of the trimer leads to an increase of replication-independent histone exchange over the coding region of transcribed genes. Taken together, these results lead to a model where Eaf5/7/3 associates with elongating polymerase to promote the disruption of nucleosomes in its path, but also their refolding in its wake. © 2014 The Authors.

  14. Advertising on mobile applications

    OpenAIRE

    Sobolevsky, Alexandr

    2015-01-01

    The article analyzes the new method of mobile advertising. Advertising in mobile applications - a subspecies of mobile marketing, where advertising is distributed using mobile phones and smartphones. Ad placement is going on inside of applications and games for smartphones. It has a high potential due to the large number of mobile phone users (over 6.5 billion in 2013).

  15. Vaccination with Leishmania infantum acidic ribosomal P0 but not with nucleosomal histones proteins controls Leishmania infantum infection in hamsters.

    Science.gov (United States)

    Pereira, Lais; Abbehusen, Melissa; Teixeira, Clarissa; Cunha, Jurema; Nascimento, Ivan P; Fukutani, Kyioshi; dos-Santos, Washington; Barral, Aldina; de Oliveira, Camila Indiani; Barral-Netto, Manoel; Soto, Manoel; Brodskyn, Cláudia Ida

    2015-02-01

    Several intracellular Leishmania antigens have been identified in order to find a potential vaccine capable of conferring long lasting protection against Leishmania infection. Histones and Acid Ribosomal proteins are already known to induce an effective immune response and have successfully been tested in the cutaneous leishmaniasis mouse model. Here, we investigate the protective ability of L. infantum nucleosomal histones (HIS) and ribosomal acidic protein P0 (LiP0) against L. infantum infection in the hamster model of visceral leishmaniasis using two different strategies: homologous (plasmid DNA only) or heterologous immunization (plasmid DNA plus recombinant protein and adjuvant). Immunization with both antigens using the heterologous strategy presented a high antibody production level while the homologous strategy immunized group showed predominantly a cellular immune response with parasite load reduction. The pcDNA-LiP0 immunized group showed increased expression ratio of IFN-γ/IL-10 and IFN-γ/TGF-β in the lymph nodes before challenge. Two months after infection hamsters immunized with the empty plasmid presented a pro-inflammatory immune response in the early stages of infection with increased expression ratio of IFN-γ/IL-10 and IFN-γ/TGF-β, whereas hamsters immunized with pcDNA-HIS presented an increase only in the ratio IFN-γ/ TGF-β. On the other hand, hamsters immunized with LiP0 did not present any increase in the IFN-γ/TGF-β and IFN-γ/IL-10 ratio independently of the immunization strategy used. Conversely, five months after infection, hamsters immunized with HIS maintained a pro-inflammatory immune response (ratio IFN-γ/ IL-10) while pcDNA-LiP0 immunized hamsters continued showing a balanced cytokine profile of pro and anti-inflammatory cytokines. Moreover we observed a significant reduction in parasite load in the spleen, liver and lymph node in this group compared with controls. Our results suggest that vaccination with L. infantum LiP0

  16. Towards elucidating the stability, dynamics and architecture of the nucleosome remodeling and deacetylase complex by using quantitative interaction proteomics.

    Science.gov (United States)

    Kloet, Susan L; Baymaz, H Irem; Makowski, Matthew; Groenewold, Vincent; Jansen, Pascal W T C; Berendsen, Madeleine; Niazi, Hassin; Kops, Geert J; Vermeulen, Michiel

    2015-05-01

    The nucleosome remodeling and deacetylase (NuRD) complex is an evolutionarily conserved chromatin-associated protein complex. Although the subunit composition of the mammalian complex is fairly well characterized, less is known about the stability and dynamics of these interactions. Furthermore, detailed information regarding protein-protein interaction surfaces within the complex is still largely lacking. Here, we show that the NuRD complex interacts with a number of substoichiometric zinc finger-containing proteins. Some of these interactions are salt-sensitive (ZNF512B and SALL4), whereas others (ZMYND8) are not. The stoichiometry of the core subunits is not affected by high salt concentrations, indicating that the core complex is stabilized by hydrophobic interactions. Interestingly, the RBBP4 and RBBP7 proteins are sensitive to high nonionic detergent concentrations during affinity purification. In a subunit exchange assay with stable isotope labeling by amino acids in cell culture (SILAC)-treated nuclear extracts, RBBP4 and RBBP7 were identified as dynamic core subunits of the NuRD complex, consistent with their proposed role as histone chaperones. Finally, using cross-linking MS, we have uncovered novel features of NuRD molecular architecture that complement our affinity purification-MS/MS data. Altogether, these findings extend our understanding of MBD3-NuRD structure and stability. MBD3 physically interacts with ZNF512B, HDAC1, ZMYND8, GATAD2B, SALL4, GATAD2A, ZNF592, MTA3, ZNF687, CDK2AP1, CHD3, ZNF532, HDAC2, MTA2, CHD4, MTA1, KPNA2, CHD5, RBBP4 and RBBP7 by pull down (View interaction) CDK2AP1 physically interacts with MBD3, MTA3, HDAC2, GATAD2A, CHD4, CDK2AP1, MTA2, HDAC1, MTA1, CHD3, GATAD2B, MBD2, RBBP4 and RBBP7 by pull down (View interaction) MBD3 physically interacts with MTA2, MTA3, RBBP4, RBBP7, HDAC2, HDAC1, CHD4, CHD3 and MTA1 by cross-linking study (View interaction). © 2014 FEBS.

  17. Mobile video with mobile IPv6

    CERN Document Server

    Minoli, Daniel

    2012-01-01

    Increased reliance on mobile devices and streaming of video content are two of the most recent changes that have led those in the video distribution industry to be concerned about the shifting or erosion of traditional advertising revenues. Infrastructure providers also need to position themselves to take advantage of these trends. Mobile Video with Mobile IPv6provides an overview of the current mobile landscape, then delves specifically into the capabilities and operational details of IPv6. The book also addresses 3G and 4G services, the application of Mobile IPv6 to streaming and other mobil

  18. Mobile shearography

    Science.gov (United States)

    Kalms, Michael; Jueptner, Werner

    2005-04-01

    By reason of their sensitivity, accuracy and non-contact as well as non-destructive characteristics, modern optical methods such as digital speckle shearography have found an increasing interest for NDT applications on the factory floor. With new carbon filter technologies and other lightweight constructions in aircraft and automotive manufacturing, adapted examination designs and especially developed testing methods are necessary. Shearography as a coherent optical method has been widely accepted as an useful NDT tool. It is a robust interferometric method to determine locations with maximum stress on various material structures. However, limitations of this technique can be found in the bulky equipment components, the interpretation of the complex sherographic result images and at the work with non-cooperative surfaces (dark absorber, bright shining reflectors). We report a mobile shearography system that was especially designed for investigations at aircraft and automotive constructions.

  19. Joint mobilization.

    Science.gov (United States)

    Saunders, Deborah Gross; Walker, J Randy; Levine, David

    2005-11-01

    Therapeutic touch has been used in human beings to soothe aches and pains. Most dogs also seem to enjoy being touched. Manual therapy techniques are skilled hand movements intended to improve tissue extensibility; increase range of motion; induce relaxation; mobilize or manipulate soft tissue and joints; modulate pain; and reduce soft tissue swelling, inflammation, or restriction. The intent of this article is to provide an overview of the principles of manual therapy, followed by selected treatment techniques for the hip, stifle, elbow, shoulder, carpus.and thoracic and lumbar spine. The techniques of G.D. Maitland, an Australian physical therapist who developed a clinically based approach in the 1960s and 1970s, are emphasized.

  20. [Social mobilization].

    Science.gov (United States)

    Bop, C

    1990-04-01

    One of the principal recommendations from Alma Ata and the Bamako Initiative was the need for communities to take responsibility for their own health--a recommendation that still remains unmet and in need of reform in Africa because of the severe economic recession and lack of resources allocated for health care in the region. The mobilization of communities "is the opposite of passivity and submission." People must demystify the notion that health care is the exclusive right of health professionals and should realize that they themselves can bring about changes from the household to the village levels; community mobilization is an integral component of development planning. African societies have developed very centralized structures requiring changes that only their own communities can bring about. Because women remain the principal agents for the family's health they should be informed, about the multiple dimensions leading to good health care to enable them to provide the rest of the family with good nourishment and health care follow-up. Children are a vulnerable and important group that require preventive care. A UNICEF experiment in Senegal is training 10-13 year old school children to visit the parents of 5 children, inform them about vaccinating their children, and to follow-up on their activities with these "adopted families." The need for short and long-term IEC interventions in Africa are a priority and effective strategies must be found to reach the majority of the rural populations where all obstacles such as the lack of infrastructure and illiteracy exist. Mali has used traditional theatre "Koteba" to reach the rural populations on a variety of health issues such as oral rehydration and diarrhea as well as the Rural Audio Library (it used cassettes rather than books) to reach villagers in their own languages. The worst obstacle facing Africa today is the refusal of officials in power to allow people to manage their own lives, of which health is a

  1. Mobile Inquiry Based Learning

    NARCIS (Netherlands)

    Specht, Marcus

    2012-01-01

    Specht, M. (2012, 8 November). Mobile Inquiry Based Learning. Presentation given at the Workshop "Mobile inquiry-based learning" at the Mobile Learning Day 2012 at the Fernuniversität Hagen, Hagen, Germany.

  2. MOBILITY: A SYSTEMS APPROACH

    Directory of Open Access Journals (Sweden)

    Mykola I. Striuk

    2015-10-01

    Full Text Available A comprehensive study on the problem of mobility in the socio-educational and technical systems was carried out: the evolution of the concept of mobility in scientific sources of XIX–XXI centuries was analyzed and the new sources on the issue of mobility introduced into scientific circulation, the interrelation of the types of mobility in the socio-pedagogical and technical systems are theoretically grounded, an integrative model of mobility in the information society is proposed. The major trends in academic mobility are identified (the transition from student mobility to mobility programs and educational services providers, the new mobility programs (franchising, double/joint degrees, combinations, nostrification etc. are characterized. The new types of mobility providers are reviewed and attention is focused on virtual universities that are now the basis of virtual mobility of students and activities which are based on the use of new ICT in higher education, especially – the Internet and mobile learning environments.

  3. Micro Mobility Marketing

    DEFF Research Database (Denmark)

    Hosbond, Jens Henrik; Skov, Mikael B.

    2008-01-01

    Mobile marketing refers to marketing of services or goods using mobile technology and mobile marketing holds potentially great economical opportunities. Traditionally, mobile marketing has been viewed as mobility in the large taking place virtually anywhere, anytime. Further, research shows...... considerable number of studies on push-based SMS mobile marketing campaigns. This paper explores a related yet different form of mobile marketing namely micro mobility marketing. Micro mobility marketing denotes mobility in the small, meaning that promotion of goods takes place within a circumscribed location......, in our case a medium-sized retail supermarket. Two prototypes based on push and pull marketing strategies are implemented and evaluated. Taking outset in a synthesis of central issues in contemporary research on mobile marketing, we discuss their role in micro mobility marketing to point to similarities...

  4. The binding of lupus-derived autoantibodies to the C-terminal peptide (83-119) of the major SmD1 autoantigen can be mediated by double-stranded DNA and nucleosomes.

    NARCIS (Netherlands)

    Dieker, J.W.C.; Bavel, C.C.A.W. van; Riemekasten, G.; Berden, J.H.M.; Vlag, J. van der

    2006-01-01

    OBJECTIVES: To evaluate the binding of lupus-derived autoantibodies, double-stranded DNA and nucleosomes to the positively charged C-terminal SmD1(residues 83-119) peptide and the full-length SmD protein. METHODS: The binding of lupus-derived monoclonal antibodies, sera from patients with systemic

  5. Abrechnung mobiler Dienste im Mobile Payment Referenzmodell

    OpenAIRE

    Pousttchi, Key

    2006-01-01

    Abrechnung mobiler Dienste im Mobile Payment Referenzmodell / K. Pousttchi, D. G. Wiedemann. - In: Handbuch E-Money, E-Payment & M-Payment / Thomas Lammer (Hrsg.). - Heidelberg : Physica-Verl., 2006. - S. 363-377

  6. Cooperating mobile robots

    Science.gov (United States)

    Harrington, John J.; Eskridge, Steven E.; Hurtado, John E.; Byrne, Raymond H.

    2004-02-03

    A miniature mobile robot provides a relatively inexpensive mobile robot. A mobile robot for searching an area provides a way for multiple mobile robots in cooperating teams. A robotic system with a team of mobile robots communicating information among each other provides a way to locate a source in cooperation. A mobile robot with a sensor, a communication system, and a processor, provides a way to execute a strategy for searching an area.

  7. The male germ cell gene regulator CTCFL is functionally different from CTCF and binds CTCF-like consensus sites in a nucleosome composition-dependent manner

    Directory of Open Access Journals (Sweden)

    Sleutels Frank

    2012-06-01

    Full Text Available Abstract Background CTCF is a highly conserved and essential zinc finger protein expressed in virtually all cell types. In conjunction with cohesin, it organizes chromatin into loops, thereby regulating gene expression and epigenetic events. The function of CTCFL or BORIS, the testis-specific paralog of CTCF, is less clear. Results Using immunohistochemistry on testis sections and fluorescence-based microscopy on intact live seminiferous tubules, we show that CTCFL is only transiently present during spermatogenesis, prior to the onset of meiosis, when the protein co-localizes in nuclei with ubiquitously expressed CTCF. CTCFL distribution overlaps completely with that of Stra8, a retinoic acid-inducible protein essential for the propagation of meiosis. We find that absence of CTCFL in mice causes sub-fertility because of a partially penetrant testicular atrophy. CTCFL deficiency affects the expression of a number of testis-specific genes, including Gal3st1 and Prss50. Combined, these data indicate that CTCFL has a unique role in spermatogenesis. Genome-wide RNA expression studies in ES cells expressing a V5- and GFP-tagged form of CTCFL show that genes that are downregulated in CTCFL-deficient testis are upregulated in ES cells. These data indicate that CTCFL is a male germ cell gene regulator. Furthermore, genome-wide DNA-binding analysis shows that CTCFL binds a consensus sequence that is very similar to that of CTCF. However, only ~3,700 out of the ~5,700 CTCFL- and ~31,000 CTCF-binding sites overlap. CTCFL binds promoters with loosely assembled nucleosomes, whereas CTCF favors consensus sites surrounded by phased nucleosomes. Finally, an ES cell-based rescue assay shows that CTCFL is functionally different from CTCF. Conclusions Our data suggest that nucleosome composition specifies the genome-wide binding of CTCFL and CTCF. We propose that the transient expression of CTCFL in spermatogonia and preleptotene spermatocytes serves to occupy a

  8. The Differential Mobilization of Histones H3.1 and H3.3 by Herpes Simplex Virus 1 Relates Histone Dynamics to the Assembly of Viral Chromatin

    Science.gov (United States)

    Conn, Kristen L.; Hendzel, Michael J.; Schang, Luis M.

    2013-01-01

    During lytic infections, HSV-1 genomes are assembled into unstable nucleosomes. The histones required for HSV-1 chromatin assembly, however, are in the cellular chromatin. We have shown that linker (H1) and core (H2B and H4) histones are mobilized during HSV-1 infection, and proposed that the mobilized histones are available for assembly into viral chromatin. However, the actual relevance of histone mobilization remained unknown. We now show that canonical H3.1 and variant H3.3 are also mobilized during HSV-1 infection. Mobilization required no HSV-1 protein expression, although immediate early or early proteins enhanced it. We used the previously known differential association of H3.3 and H3.1 with HSV-1 DNA to test the relevance of histone mobilization. H3.3 binds to HSV-1 genomes first, whereas H3.1 only binds after HSV-1 DNA replication initiates. Consistently, H3.3 and H3.1 were differentially mobilized. H3.1 mobilization decreased with HSV-1 DNA replication, whereas H3.3 mobilization was largely unaffected by it. These results support a model in which previously mobilized H3.1 is immobilized by assembly into viral chromatin during HSV-1 DNA replication, whereas H3.3 is mobilized and assembled into HSV-1 chromatin throughout infection. The differential mobilizations of H3.3 and H3.1 are consistent with their differential assembly into viral chromatin. These data therefore relate nuclear histone dynamics to the composition of viral chromatin and provide the first evidence that histone mobilization relates to viral chromatin assembly. PMID:24130491

  9. TYPOLOGIES OF MOBILE APPLICATIONS

    OpenAIRE

    Ion Ivan; Alin Zamfiroiu; Dragoş Palaghiţă3

    2013-01-01

    Mobile applications and their particularities are analyzed. Mobile application specific characteristics are defined. Types of applications are identified and analyzed. The paper established differences between mobile applications and mobile application categories. For each identified type the specific structures and development model are identified.

  10. Next generation mobile broadcasting

    CERN Document Server

    Gómez-Barquero, David

    2013-01-01

    Next Generation Mobile Broadcasting provides an overview of the past, present, and future of mobile multimedia broadcasting. The first part of the book-Mobile Broadcasting Worldwide-summarizes next-generation mobile broadcasting technologies currently available. This part covers the evolutions of the Japanese mobile broadcasting standard ISDB-T One-Seg, ISDB-Tmm and ISDB-TSB; the evolution of the South Korean T-DMB mobile broadcasting technology AT-DMB; the American mobile broadcasting standard ATSC-M/H; the Chinese broadcasting technologies DTMB and CMMB; second-generation digital terrestrial

  11. Mobile platform security

    CERN Document Server

    Asokan, N; Dmitrienko, Alexandra

    2013-01-01

    Recently, mobile security has garnered considerable interest in both the research community and industry due to the popularity of smartphones. The current smartphone platforms are open systems that allow application development, also for malicious parties. To protect the mobile device, its user, and other mobile ecosystem stakeholders such as network operators, application execution is controlled by a platform security architecture. This book explores how such mobile platform security architectures work. We present a generic model for mobile platform security architectures: the model illustrat

  12. Charging of mobile services by mobile payment reference model

    OpenAIRE

    Pousttchi, Key; Wiedemann, Dietmar Georg

    2005-01-01

    The purpose of the paper is to analyze mobile payments in the mobile commerce scenario. Therefore, we first classify the mobile payment in the mobile commerce scenario by explaining general offer models, charging concepts, and intermediaries. Second, we describe the mobile payment reference model, especially, the mobile payment reference organization model and different mobile payment standard types. Finally, we conclude our findings.

  13. Abrechnung mobiler Dienste im Mobile-Payment-Referenzmodell

    OpenAIRE

    Pousttchi, Key; Wiedemann, Dietmar Georg

    2005-01-01

    The purpose of the paper is to analyze mobile payments in the mobile commerce scenario. Therefore, we first classify the mobile payment in the mobile commerce scenario by explaining general offer models, charging concepts, and intermediaries. Second, we describe the mobile payment reference model, especially, the mobile payment reference organization model and different mobile payment standard types. Finally, we conclude our findings.

  14. MOBILE TELECOMMUNICATIONS SERVICES AND MOBILE HEALTH DEVICES

    OpenAIRE

    Gheorghe Meghisan; Georgeta-Madalina Meghisan

    2015-01-01

    The purpose of this research paper is to identify the potential of mobile health devices with a positive impact on the public health care system from Romania. More people monitoring their health situation with the help of mobile health applications could lead to less money spent by the public health sector with treating more advanced diseases. Approach/ methodology. The analysis of the Romanian mobile telecommunications market and health situation of the population from Romania was based on s...

  15. Mobile Portal Implementation Strategy

    DEFF Research Database (Denmark)

    Gao, Ping; Damsgaard, Jan

    2005-01-01

    Mobile portal plays an important role in mobile commerce market. Current literature focuses on static analysis on the value chain of mobile portals. This article provides a dynamic perspective on mobile portal strategy. Drawing upon network economics, we describe mobile portal implementation...... as a fourphase process. In different phase, a portal provider has various challenges to overcome and adopt diverse strategies, and correspondingly the regulator has different foci. The conceptual framework proposed in this article offers a basis for further analyses on the market dynamics of mobile commerce......, and can be generalized to studying other networked technologies...

  16. Mobile Router Technology Development

    Science.gov (United States)

    Ivancic, William D.; Stewart, David H.; Bell, Terry L.; Kachmar, Brian A.; Shell, Dan; Leung, Kent

    2002-01-01

    Cisco Systems and NASA have been performing joint research on mobile routing technology under a NASA Space Act Agreement. Cisco developed mobile router technology and provided that technology to NASA for applications to aeronautic and space-based missions. NASA has performed stringent performance testing of the mobile router, including the interaction of routing and transport-level protocols. This paper describes mobile routing, the mobile router, and some key configuration parameters. In addition, the paper describes the mobile routing test network and test results documenting the performance of transport protocols in dynamic routing environments.

  17. Head First Mobile Web

    CERN Document Server

    Gardner, Lyza; Grigsby, Jason

    2011-01-01

    Despite the huge number of mobile devices and apps in use today, your business still needs a website. You just need it to be mobile. Head First Mobile Web walks you through the process of making a conventional website work on a variety smartphones and tablets. Put your JavaScript, CSS media query, and HTML5 skills to work-then optimize your site to perform its best in the demanding mobile market. Along the way, you'll discover how to adapt your business strategy to target specific devices. Navigate the increasingly complex mobile landscapeTake both technical and strategic approaches to mobile

  18. 33 CFR 165.835 - Security Zone; Port of Mobile, Mobile Ship Channel, Mobile, AL.

    Science.gov (United States)

    2010-07-01

    ..., Mobile Ship Channel, Mobile, AL. 165.835 Section 165.835 Navigation and Navigable Waters COAST GUARD... § 165.835 Security Zone; Port of Mobile, Mobile Ship Channel, Mobile, AL. (a) Definition. As used in... following areas are security zones: all waters of the Port of Mobile and Mobile Ship Channel— (1) Within 100...

  19. Exploring the Mobility of Mobile Phone Users

    CERN Document Server

    Csáji, Balázs Cs; Traag, V A; Delvenne, Jean-Charles; Huens, Etienne; Van Dooren, Paul; Smoreda, Zbigniew; Blondel, Vincent D

    2013-01-01

    Mobile phone datasets allow for the analysis of human behavior on an unprecedented scale. The social network, temporal dynamics and mobile behavior of mobile phone users have often been analyzed independently from each other using mobile phone datasets. In this article, we explore the connections between various features of human behavior extracted from a large mobile phone dataset. Our observations are based on the analysis of communication data of 100000 anonymized and randomly chosen individuals in a dataset of communications in Portugal. We show that clustering and principal component analysis allow for a significant dimension reduction with limited loss of information. The most important features are related to geographical location. In particular, we observe that most people spend most of their time at only a few locations. With the help of clustering methods, we then robustly identify home and office locations and compare the results with official census data. Finally, we analyze the geographic spread ...

  20. Tandem mobile robot system

    Science.gov (United States)

    Buttz, James H.; Shirey, David L.; Hayward, David R.

    2003-01-01

    A robotic vehicle system for terrain navigation mobility provides a way to climb stairs, cross crevices, and navigate across difficult terrain by coupling two or more mobile robots with a coupling device and controlling the robots cooperatively in tandem.

  1. Mobile IP: Security & application

    NARCIS (Netherlands)

    Tuquerres, G.; Salvador, M.R.; Sprenkels, Ron

    1999-01-01

    As required in the TGS Mobile IP Advanced Module, this paper presents a survey of common security threats which mobile IP networks are exposed to as well as some proposed solutions to deal with such threats.

  2. Seasonality, mobility, and livability.

    Science.gov (United States)

    2012-01-31

    Signature project 4a, Seasonality, Mobility, and Livability investigated the effects of weather, season, built environment, community amenities, attitudes, and demographics on mobility and quality of life (QOL). A four season panel survey exami...

  3. Ion mobility sensor system

    Science.gov (United States)

    Xu, Jun; Watson, David B.; Whitten, William B.

    2013-01-22

    An ion mobility sensor system including an ion mobility spectrometer and a differential mobility spectrometer coupled to the ion mobility spectrometer. The ion mobility spectrometer has a first chamber having first end and a second end extending along a first direction, and a first electrode system that generates a constant electric field parallel to the first direction. The differential mobility spectrometer includes a second chamber having a third end and a fourth end configured such that a fluid may flow in a second direction from the third end to the fourth end, and a second electrode system that generates an asymmetric electric field within an interior of the second chamber. Additionally, the ion mobility spectrometer and the differential mobility spectrometer form an interface region. Also, the first end and the third end are positioned facing one another so that the constant electric field enters the third end and overlaps the fluid flowing in the second direction.

  4. Innovative island mobile vet.

    Science.gov (United States)

    Forster, Dan

    2016-06-11

    One of the UK's first mobile veterinary clinics was recently awarded a Queen's Award for Innovation. Mobile Vet was launched on the Isle of Wight in 2013 by Dan Forster and his wife Kirsty, a veterinary nurse. British Veterinary Association.

  5. Ion mobility spectrometry

    CERN Document Server

    Eiceman, GA

    2005-01-01

    Key Developments for Faster, More Precise Detection Capabilities Driven by the demand for the rapid and advanced detection of explosives, chemical and biological warfare agents, and narcotics, ion mobility spectrometry (IMS) undergone significant refinements in technology, computational capabilities, and understanding of the principles of gas phase ion chemistry and mobility. Beginning with a thorough discussion of the fundamental theories and physics of ion mobility, Ion Mobility Spectrometry, Second Edition describes the recent advances in instrumentation and newly

  6. Staging interrail mobilities

    DEFF Research Database (Denmark)

    Jensen, Martin Trandberg; Gyimóthy, Szilvia; Jensen, Ole B.

    2016-01-01

    This article applies the multiscalar ‘staging mobilities’ framework from the emergent subfield of mobilities design to analyse an enduring European rail travel phenomenon, interrail. This discussion extends and contributes to tourism mobilities research. Second, the article enriches previous...... and seat reservations. To reach these aims, the research design intertwines multi-sited ethnography, netnography, survey and interviews. The conclusion offers theoretical reflections pertaining to the role of mobilities designs and methodical hybrids in tourism mobilities research....

  7. Fixed mobile convergence handbook

    CERN Document Server

    Ahson, Syed A

    2010-01-01

    From basic concepts to future directions, this handbook provides technical information on all aspects of fixed-mobile convergence (FMC). The book examines such topics as integrated management architecture, business trends and strategic implications for service providers, personal area networks, mobile controlled handover methods, SIP-based session mobility, and supervisory and notification aggregator service. Case studies are used to illustrate technical and systematic implementation of unified and rationalized internet access by fixed-mobile network convergence. The text examines the technolo

  8. Social Perspectives on Mobility

    DEFF Research Database (Denmark)

    Uth Thomsen, Thyra; Gudmundsson, Henrik; Drewes Nielsen, Lise

    interdisciplinary theoretical approaches with empirical case studies analysing and appraising innovative policies from Scandinavia, this volume demonstrates that mobility research is a key issue within social enquiry. It addresses three broad themes. Firstly, mobility as a constructed social reality, examining how...... for both research and policy. In the final section of the book new visions for research into sustainability and mobility are laid out....

  9. Mobile Student Information System

    Science.gov (United States)

    Asif, Muhammad; Krogstie, John

    2011-01-01

    Purpose: A mobile student information system (MSIS) based on mobile computing and context-aware application concepts can provide more user-centric information services to students. The purpose of this paper is to describe a system for providing relevant information to students on a mobile platform. Design/methodology/approach: The research…

  10. Mobile energy sharing futures

    DEFF Research Database (Denmark)

    Worgan, Paul; Knibbe, Jarrod; Plasencia, Diego Martinez

    2016-01-01

    We foresee a future where energy in our mobile devices can be shared and redistributed to suit our current task needs. Many of us are beginning to carry multiple mobile devices and we seek to re-evaluate the traditional view of a mobile device as only accepting energy. In our vision, we can...... sharing futures....

  11. Distributed mobility management - framework & analysis

    NARCIS (Netherlands)

    Liebsch, M.; Seite, P.; Karagiannis, Georgios

    Mobile operators consider the distribution of mobility anchors to enable offloading some traffic from their core network. The Distributed Mobility Management (DMM) Working Group is investigating the impact of decentralized mobility management to existing protocol solutions, while taking into account

  12. Origin replication complex binding, nucleosome depletion patterns, and a primary sequence motif can predict origins of replication in a genome with epigenetic centromeres.

    Science.gov (United States)

    Tsai, Hung-Ji; Baller, Joshua A; Liachko, Ivan; Koren, Amnon; Burrack, Laura S; Hickman, Meleah A; Thevandavakkam, Mathuravani A; Rusche, Laura N; Berman, Judith

    2014-09-02

    Origins of DNA replication are key genetic elements, yet their identification remains elusive in most organisms. In previous work, we found that centromeres contain origins of replication (ORIs) that are determined epigenetically in the pathogenic yeast Candida albicans. In this study, we used origin recognition complex (ORC) binding and nucleosome occupancy patterns in Saccharomyces cerevisiae and Kluyveromyces lactis to train a machine learning algorithm to predict the position of active arm (noncentromeric) origins in the C. albicans genome. The model identified bona fide active origins as determined by the presence of replication intermediates on nondenaturing two-dimensional (2D) gels. Importantly, these origins function at their native chromosomal loci and also as autonomously replicating sequences (ARSs) on a linear plasmid. A "mini-ARS screen" identified at least one and often two ARS regions of ≥100 bp within each bona fide origin. Furthermore, a 15-bp AC-rich consensus motif was associated with the predicted origins and conferred autonomous replicating activity to the mini-ARSs. Thus, while centromeres and the origins associated with them are epigenetic, arm origins are dependent upon critical DNA features, such as a binding site for ORC and a propensity for nucleosome exclusion. DNA replication machinery is highly conserved, yet the definition of exactly what specifies a replication origin differs in different species. Here, we utilized computational genomics to predict origin locations in Candida albicans by combining locations of binding sites for the conserved origin replication complex, necessary for replication initiation, together with chromatin organization patterns. We identified predicted sequences that exhibited bona fide origin function and developed a linear plasmid assay to delimit the DNA fragments necessary for origin function. Additionally, we found that a short AC-rich motif, which is enriched in predicted origins, is required for

  13. Mobility Charters and Manifestos

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    2010-01-01

    and ethical positions on mobility prevail. Such ‘imagined correct mobility behavior’ are drawing on larger issues of societal change that need to be brought out in a critical analysis and discussion reflecting the attempts to control, design and orchestrate mobility patterns. The paper therefore argues within......This paper explore a number of different cases of articulating notions of ‘correct’ mobility behavior and practice by looking into charters, manifestos and codes of mobility regulation. Within such discourses of ‘correct mobility’ more or less subtle expressions of power as well as normative...

  14. Mobile Virtual Private Networking

    Science.gov (United States)

    Pulkkis, Göran; Grahn, Kaj; Mårtens, Mathias; Mattsson, Jonny

    Mobile Virtual Private Networking (VPN) solutions based on the Internet Security Protocol (IPSec), Transport Layer Security/Secure Socket Layer (SSL/TLS), Secure Shell (SSH), 3G/GPRS cellular networks, Mobile IP, and the presently experimental Host Identity Protocol (HIP) are described, compared and evaluated. Mobile VPN solutions based on HIP are recommended for future networking because of superior processing efficiency and network capacity demand features. Mobile VPN implementation issues associated with the IP protocol versions IPv4 and IPv6 are also evaluated. Mobile VPN implementation experiences are presented and discussed.

  15. Engaging with mobile methods

    DEFF Research Database (Denmark)

    Jensen, Martin Trandberg

    2014-01-01

    This chapter showcases how mobile methods are more than calibrated techniques awaiting application by tourism researchers, but productive in the enactment of the mobile (Law and Urry, 2004). Drawing upon recent findings deriving from a PhD course on mobility and mobile methods it reveals...... the conceptual ambiguousness of the term ‘mobile methods’. In order to explore this ambiguousness the chapter provides a number of examples deriving from tourism research, to explore how mobile methods are always entangled in ideologies, predispositions, conventions and practice-realities. Accordingly......, the engagements with methods are acknowledged to be always political and contextual, reminding us to avoid essentialist discussions regarding research methods. Finally, the chapter draws on recent fieldwork to extend developments in mobilities-oriented tourism research, by employing auto-ethnography to call...

  16. Embodied Cultures of Mobilities

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    2010-01-01

    and material artifacts. The paper target the complex relationship between the moving, sensing body and the material and built environment of infrastructures and mobility modes in order to explore what norms, and meanings, and everyday life mobility cultures are being produced and re-produced in this process...... contemporary theorists understanding bodily movement, material sites of mobility, and social interactions must be consulted along the road (e.g. Latour’s work on objects and ANT, Thrift’s work on the body and ‘non-representational theory, and Massumi’s notions of affects and emotions related to bodily mobility......The paper explores the relationship between the body and mobility by looking into a number of modes of transportation and their ways of constructing particular engagements with mobility. The ‘mobile embodiments’ are significant to a material and symbolic set of relations between human agents...

  17. European mobility cultures

    DEFF Research Database (Denmark)

    Haustein, Sonja; Nielsen, Thomas A. Sick

    2016-01-01

    More targeted European policies promoting green travel patterns require better knowledge on differing mobility cultures across European regions. As a basis for this, we clustered the EU population into eight mobility styles based on Eurobarometer data. The mobility styles - including, for example......-economic resources. In a second step, the 28 EU member countries were clustered into six country clusters based on their representation of mobility styles. The country clusters indicate the existence of considerably different mobility cultures across the EU. Sub-regions can be identified that have highly different...... positions on the path towards sustainable mobility and therefore different requirements towards European platforms and support measures, e.g. for 'Sustainable Urban Mobility Plans'. The country clusters can provide a starting point for future communication and targeting of European efforts in sustainable...

  18. A different glimpse into mobilities : On the interrelations between daily spatial mobility and social mobility

    OpenAIRE

    Fortunati, Leopoldina; Taipale, Sakari

    2017-01-01

    We explore the link between daily spatial mobility and social mobility, taking changes in the contemporary labor market and family as examples. We propose a new theoretical approach to mobility that is defined as a productive force of social labor. Analyzing the relationship between daily spatial mobility and social mobility, we show that spatial mobility has become a strategy of compensation for the lack of social mobility. Explanations for the increase in daily spatial mobility are also pro...

  19. Genome-wide screen of cell-cycle regulators in normal and tumor cells identifies a differential response to nucleosome depletion.

    Science.gov (United States)

    Sokolova, Maria; Turunen, Mikko; Mortusewicz, Oliver; Kivioja, Teemu; Herr, Patrick; Vähärautio, Anna; Björklund, Mikael; Taipale, Minna; Helleday, Thomas; Taipale, Jussi

    2017-01-17

    To identify cell cycle regulators that enable cancer cells to replicate DNA and divide in an unrestricted manner, we performed a parallel genome-wide RNAi screen in normal and cancer cell lines. In addition to many shared regulators, we found that tumor and normal cells are differentially sensitive to loss of the histone genes transcriptional regulator CASP8AP2. In cancer cells, loss of CASP8AP2 leads to a failure to synthesize sufficient amount of histones in the S-phase of the cell cycle, resulting in slowing of individual replication forks. Despite this, DNA replication fails to arrest, and tumor cells progress in an elongated S-phase that lasts several days, finally resulting in death of most of the affected cells. In contrast, depletion of CASP8AP2 in normal cells triggers a response that arrests viable cells in S-phase. The arrest is dependent on p53, and preceded by accumulation of markers of DNA damage, indicating that nucleosome depletion is sensed in normal cells via a DNA-damage -like response that is defective in tumor cells.

  20. hOGG1 removes solution-accessible 8-oxoG lesions from globally-substituted nucleosomes except at the dyad region.

    Science.gov (United States)

    Bilotti, Katharina; Tarantino, Mary E; Delaney, Sarah

    2018-01-17

    Persistent DNA damage is responsible for mutagenesis, aging, and disease. Repair of the prototypic oxidatively damaged guanine lesion 8-oxo-7,8-dihydroguanine (8-oxoG) is initiated by oxoguanine glycosylase (hOGG1 in humans). In this work, we examine hOGG1 activity on DNA packaged as it is in chromatin, in a nucleosome core particle (NCP). We use synthetic methods to generate a population of NCPs with G to 8-oxoG substitutions and evaluate the global profile of hOGG1 repair in packaged DNA. For several turns of the helix, we observe that solution-accessible 8-oxoG are sites of activity for hOGG1. At the dyad axis, however, hOGG1 activity is suppressed, even at lesions predicted to be solution accessible by hydroxyl radical footprinting (HRF). We predict this diminished activity is due to properties of the DNA unique to the dyad axis and/or the local histone environment. In contrast to the dyad axis, the DNA ends reveal hOGG1 activity at sites predicted by HRF to be both solution accessible and inaccessible. We attribute the lack of correlation between hOGG1 activity and solution accessibility at the ends of the DNA to transient unwrapping of the DNA from the protein core, thus exposing the inward-facing lesions.

  1. The presence of chimeric DNA consisting of 5' regions of the hemoglobin and nucleosome assembly protein-1 genes in Paramecium caudatum macronuclear genomic DNA.

    Science.gov (United States)

    Nishiyama, Norihito; Mikami, Kazuyuki; Ochiai, Takehiko; Yamauchi, Kiyoshi

    2009-04-01

    We detected an unexpected small-sized DNA fragment during polymerase chain reaction (PCR) analysis of the heterogeneity of a macronuclear intergenic region of Paramecium caudatum. Southern blotting of total genomic DNA with the PCR product as a probe indicated that the small-sized DNA fragment constituted part of the macronuclear genome. Sequencing revealed that the PCR product was a chimeric DNA structure that may be generated by tail-to-tail fusion of the 5' region of the hemoglobin (hb) gene to most of the nucleosome assembly protein-1 (nap-1) gene. Short tandem repeats consisting of tetra- and tri-nucleotides exist at the putative cleavage sites in the hb and nap-1 genes, respectively. This feature differs from those found at the boundaries of TA-internal eliminated sequences in the P. aurelia complex and at transposable elements in other species. This suggests that the chimeric DNA is generated by a novel mechanism. Although the chimeric DNA contains the hb and nap-1 promoters, transcripts corresponding to the chimeric DNA were not detected by reverse transcription (RT)-PCR analysis during vegetative cell growth. Possible roles of chimeric DNA are discussed.

  2. Mobile healthcare informatics.

    Science.gov (United States)

    Siau, Keng; Shen, Zixing

    2006-06-01

    Advances in wireless technology give pace to the rapid development of mobile applications. The coming mobile revolution will bring dramatic and fundamental changes to our daily life. It will influence the way we live, the way we do things, and the way we take care of our health. For the healthcare industry, mobile applications provide a new frontier in offering better care and services to patients, and a more flexible and mobile way of communicating with suppliers and patients. Mobile applications will provide important real time data for patients, physicians, insurers, and suppliers. In addition, it will revolutionalize the way information is managed in the healthcare industry and redefine the doctor - patient communication. This paper discusses different aspects of mobile healthcare. Specifically, it presents mobile applications in healthcare, and discusses possible challenges facing the development of mobile applications. Obstacles in developing mobile healthcare applications include mobile device limitations, wireless networking problems, infrastructure constraints, security concerns, and user distrust. Research issues in resolving or alleviating these problems are also discussed in the paper.

  3. Frailty and Mobility.

    Science.gov (United States)

    Eeles, Eamonn; Low Choy, Nancy

    2015-01-01

    Frailty represents a state of heightened vulnerability. Mobility impairment contributes to the construct of frailty and channels adverse events. While mobility disorder is universal at a high burden of frailty, neither mobility nor balance dysfunction is sufficient to fully define frailty. Frailty represents proximity to complex system failure, with higher-order disturbance, such as mobility and balance disturbance, as a consequence. Impairment of mobility and balance is a common manifestation of illness in the frail individual and is therefore a sensitive marker of acute disease, putatively also in delirium. Clinical measurement of mobility and balance should be prioritized. Consequently, assessment tools, such as the de Morton Mobility Index and the Hierarchical Assessment of Balance and Mobility, are being explored, with the sensitivity of the latter illustrated in the acute hospital setting. Walking with speed and under dual/multi-task conditions better differentiates healthier and frail ambulant adults, providing a basis for screening older adults for pre-emptive interventions. Specific mobility and balance interventions reduce falls risk. However, patients with dementia walk too fast for their level of frailty, creating an ethical dimension to rehabilitation and risk. Overall, there is no need for reduced mobility to reinforce the frailty stereotype; both are potentially modifiable and amenable to intervention strategies. 2015 S. Karger AG, Basel.

  4. Social Perspectives on Mobility

    DEFF Research Database (Denmark)

    Uth Thomsen, Thyra; Gudmundsson, Henrik; Drewes Nielsen, Lise

    interdisciplinary theoretical approaches with empirical case studies analysing and appraising innovative policies from Scandinavia, this volume demonstrates that mobility research is a key issue within social enquiry. It addresses three broad themes. Firstly, mobility as a constructed social reality, examining how......Globalisation is heavily dependent on physical transport, as people and goods travel over longer distances and with higher frequency. Movement and mobility have become integrated parts of late modern identity and practice, and a state of flux can be sensed everywhere. Bringing together the latest...... individuals construct notions of mobility in their everyday life and practice. Secondly, mobility as spatial co-ordination and transgression, and finally, mobility as a policy theme, where the contributors explore recent developments in transport policy at national and European levels, suggesting ways forward...

  5. Social Perspectives on Mobility

    DEFF Research Database (Denmark)

    Uth Thomsen, Thyra; Gudmundsson, Henrik; Drewes Nielsen, Lise

    Globalisation is heavily dependent on physical transport, as people and goods travel over longer distances and with higher frequency. Movement and mobility have become integrated parts of late modern identity and practice, and a state of flux can be sensed everywhere. Bringing together the latest...... interdisciplinary theoretical approaches with empirical case studies analysing and appraising innovative policies from Scandinavia, this volume demonstrates that mobility research is a key issue within social enquiry. It addresses three broad themes. Firstly, mobility as a constructed social reality, examining how...... individuals construct notions of mobility in their everyday life and practice. Secondly, mobility as spatial co-ordination and transgression, and finally, mobility as a policy theme, where the contributors explore recent developments in transport policy at national and European levels, suggesting ways forward...

  6. Mens mobile health

    DEFF Research Database (Denmark)

    Levisen, Vinie Diana Hvidbak; Castaño, Francisco Mansilla; Jensen, Camilla Skovbjerg

    2016-01-01

    health applications in relation to fitness, heart rate, blood pressure, BMI, fat and muscle mass, BodyAge? Main messages: A CCT, that contributes with knowledge of how mobile health applications are impacting the physical activity levels among men with little or no education and frequency of how often......Abstract til: 9th European Public Health Conference – All for health – Health for All 9 – 12 November 2016. Danish title: Mænds mobile sundhed English title: Men mobile health Authors: Vinie Diana Hvidbak Levisen, Associated professor, RN, MLP Knowledge Center for Health Promotion University...... Preferences Keywords: Men with little or no education, physical health, mobile health application. Types of presentations: First presentation preference: E-Poster presentations Second presentation preference: Pitch presentation Abstract Background: Men mobile health contributes knowledge of how mobile health...

  7. Talk, Mobility and Materialities

    DEFF Research Database (Denmark)

    McIlvenny, Paul

    The intersection of the quotidian practices of social interaction, learning and mobility outside of the classroom – for example, the ways in which talk shapes how children learn to be actively mobile – has been little studied until recently. This paper develops a social interactional approach...... to analysing talk and mobile action in what are arguably two quintessentially Nordic mobility practices, namely cycling and skiing. More specifically the focus is on investigating and comparing how a child learns to cycle in a bike-friendly urban infrastructure, and how a child learns to ski cross...... locates similarities and differences in how specific materialities are made salient in interactional practices. For example, caregivers talk about weather and surface conditions as a resource for instructed mobile action. Also, a route, a trajectory or a line of movement in a mobile formation...

  8. CERN Mobility Survey

    CERN Multimedia

    GS Department

    2011-01-01

    The Institute of Shipping and Transport of the University of the Aegean and the National Technical University of Athens are partners with CERN in a study of mobility patterns between and within the CERN sites and to that effect have realized a mobility survey dedicated to the CERN community.         The study aims to understand: How you presently get around the CERN sites; What problems you encounter regarding mobility; What your needs are; What improvements you’d like to see; What measures you would like to see implemented most. The replies we receive will enable us to define a general policy promoting the diversity of mobility at CERN and to establish and quantify the strategic actions to be implemented for both the short and medium term. The objectives of the transport mobility plans are to: Facilitate mobility within and between the CERN sites by identifying adequate solutions in response to individual ...

  9. Quality attributes for mobile applications

    OpenAIRE

    Fernandes, João M.; Ferreira, André Leite

    2016-01-01

    A mobile application is a type of software application developed to run on a mobile device. The chapter discusses the main characteristics of mobile devices, since they have a great impact on mobile applications. It also presents the classification of mobile applications according to two main types: native and web-based applications. Finally, this chapter identifies the most relevant types of quality attributes for mobile applications. It shows that the relevant quality attributes for mobile ...

  10. MobiLED: mobile-led and leading via mobile

    CSIR Research Space (South Africa)

    Ford, M

    2009-03-01

    Full Text Available the past three years it has become evident that many of the initiative's innovations have a wider application than originally envisaged. This paper will discuss the results of the education-related MobiLED pilots and expands on the possibilities of using...

  11. Mobile learning in medicine

    Science.gov (United States)

    Serkan Güllüoüǧlu, Sabri

    2013-03-01

    This paper outlines the main infrastructure for implicating mobile learning in medicine and present a sample mobile learning application for medical learning within the framework of mobile learning systems. Mobile technology is developing nowadays. In this case it will be useful to develop different learning environments using these innovations in internet based distance education. M-learning makes the most of being on location, providing immediate access, being connected, and acknowledges learning that occurs beyond formal learning settings, in places such as the workplace, home, and outdoors. Central to m-learning is the principle that it is the learner who is mobile rather than the device used to deliver m learning. The integration of mobile technologies into training has made learning more accessible and portable. Mobile technologies make it possible for a learner to have access to a computer and subsequently learning material and activities; at any time and in any place. Mobile devices can include: mobile phone, personal digital assistants (PDAs), personal digital media players (eg iPods, MP3 players), portable digital media players, portable digital multimedia players. Mobile learning (m-learning) is particularly important in medical education, and the major users of mobile devices are in the field of medicine. The contexts and environment in which learning occurs necessitates m-learning. Medical students are placed in hospital/clinical settings very early in training and require access to course information and to record and reflect on their experiences while on the move. As a result of this paper, this paper strives to compare and contrast mobile learning with normal learning in medicine from various perspectives and give insights and advises into the essential characteristics of both for sustaining medical education.

  12. Mobile Entrepreneurship and Employment

    OpenAIRE

    Imaizumi, Saori; Andjelkovic, Maja

    2012-01-01

    Given its strong recent growth, the global mobile industry is now a major source of employment opportunities. Mobile industry jobs can be categorized into direct and indirect ones, with a diverse labor force supplying each category. Direct jobs are created by mobile operators and manufacturers in professions that range from engineers to managers to sales support staff. The International Telecommunication Union (ITU) estimates that around 1.5 million people are directly employed in the industr...

  13. Enhancing mobile learning security

    OpenAIRE

    Shonola, Shaibu Adekunle; Joy, Mike

    2016-01-01

    Mobile devices have been playing vital roles in modern day education delivery as students can access or download learning materials on their smartphones and tablets, they can also install educational apps and study anytime, anywhere. The need to provide adequate security for portable devices being used for learning cannot be underestimated. In this paper, we present a mobile security enhancement app, designed and developed for Android smart mobile devices in order to promote security awarenes...

  14. Mobile lifestyles : Conceptualizing heterogeneity in mobile youth culture

    NARCIS (Netherlands)

    Vanden Abeele, M.M.P.

    2014-01-01

    Adolescents’ characteristic understanding and use of mobile phones have led observers to speak of a “mobile youth culture.” This article explores whether we can differentiate lifestyles within mobile youth culture. We construct a user typology of Flemish adolescent mobile phone users based on mobile

  15. The setup of a mobile mobility panel for the Netherlands

    NARCIS (Netherlands)

    Geurs, Karst Teunis; Veenstra, Sander; Thomas, Tom

    2013-01-01

    This paper describes the setup of the Dutch Mobile Mobility Panel project, in which GPS-enabled mobile phones (smartphones) are used as a passive multiple-week and multiple-year travel behaviour data collection tool. The data collection methodology used in the Dutch Mobile Mobility Panel comprised

  16. Mobile Test Capabilities

    Data.gov (United States)

    Federal Laboratory Consortium — The Electrical Power Mobile Test capabilities are utilized to conduct electrical power quality testing on aircraft and helicopters. This capability allows that the...

  17. Test for mobility

    Directory of Open Access Journals (Sweden)

    Li Kheng

    2014-06-01

    Full Text Available Mobile application testing is a process by which application software developed for mobile devices is tested, in aspects as functionality, usability and consistency. These applications can be pre-installed or installed from distribution platforms for mobile software. Due mobile devices have witnessed of a phenomenal growth in recent years (a study by Yankee Group predicted to generate $ 4.2 billion in revenue for 2013 through 7000 million downloads of applications the U.S., it is necesary test engineers increasingly specialize. In this article are described the types of applications, the challenges of testing, and its automation.

  18. Mobile Solar Tracker Facility

    Data.gov (United States)

    Federal Laboratory Consortium — NIST's mobile solar tracking facility is used to characterize the electrical performance of photovoltaic panels. It incorporates meteorological instruments, a solar...

  19. Mobile computing handbook

    CERN Document Server

    Ilyas, Mohammad

    2004-01-01

    INTRODUCTION AND APPLICATIONS OF MOBILE COMPUTING Wearable Computing,A. Smailagic and D.P. Siewiorek Developing Mobile Applications: A Lime Primer,G.P. Picco, A.L. Murphy, and G.-C. Roman Pervasive Application Development: Approaches and Pitfalls,G. Banavar, N. Cohen, and D. Soroker ISAM, Joining Context-Awareness and Mobility to Building Pervasive Applications,I. Augustin, A. Corrêa Yamin, J.L. Victória Barbosa, L. Cavalheiro da Silva, R. Araújo Real, G. Frainer, G.G. Honrich Cavalheiro, and C.F. Resin Geyer Integrating Mobile Wireless Devices into the Computational Grid,T. Phan, L. Huan

  20. Understanding pastoral mobility

    DEFF Research Database (Denmark)

    Adriansen, Hanne Kirstine

    2008-01-01

    Based on a case study from Sahelian Senegal, this paper analyses how various actors perceive the importance of pastoral mobility and presents issues of importance for understanding the use of mobility among Fulani of Ferlo. One knowledge system is a scientific one, the 'new rangeland paradigm......'. According to this, pastoral mobility is a means to balance variability in dryland resources; hence, 'nature' is the point of departure. Another knowledge system is local pastoral knowledge. For the pastoralists, the well-being of their animals is the point of departure and mobility is used to ensure...... and how this may change in the future....

  1. Practical mobile forensics

    CERN Document Server

    Bommisetty, Satish; Mahalik, Heather

    2014-01-01

    The book is an easy-to-follow guide with clear instructions on various mobile forensic techniques. The chapters and the topics within are structured for a smooth learning curve, which will swiftly empower you to master mobile forensics. If you are a budding forensic analyst, consultant, engineer, or a forensic professional wanting to expand your skillset, this is the book for you. The book will also be beneficial to those with an interest in mobile forensics or wanting to find data lost on mobile devices. It will be helpful to be familiar with forensics in general but no prior experience is re

  2. Mobile Project Management

    Directory of Open Access Journals (Sweden)

    Catalin BOJA

    2006-01-01

    Full Text Available Based on the development of new communication and data transfer technologies, the mobile solutions for the management process have been able to provide new ways to conduct management actions. This environment describes methods and tools available only here, which will bring information, speed and efficiency to any stage and component of the management process. The paper takes into discussion the impact of the technological development on the management process paradigm. The paper presents the main aspects regarding the business and management models used in mobile management. The role of mobile multimedia informatics applications in mobile management is highlighted.

  3. Programming the Mobile Web

    CERN Document Server

    Firtman, Maximiliano

    2010-01-01

    Today's market for mobile apps goes beyond the iPhone to include BlackBerry, Nokia, Windows Phone, and smartphones powered by Android, webOS, and other platforms. If you're an experienced web developer, this book shows you how to build a standard app core that you can extend to work with specific devices. You'll learn the particulars and pitfalls of building mobile apps with HTML, CSS, and other standard web tools. You'll also explore platform variations, finicky mobile browsers, Ajax design patterns for mobile, and much more. Before you know it, you'll be able to create mashups using Web 2.

  4. Guest Editorial ~ Mobile Learning

    Directory of Open Access Journals (Sweden)

    Mohamed Ally

    2007-06-01

    Full Text Available This special IRRODL issue on mobile learning is timely because of the proliferation of mobile technology in society, globalization, and the need to re-examine how learning materials are designed and delivered for the new generation of learners. In today’s world, people are on the move and are demanding access to learning materials and information anytime and anywhere. At the same time, there is increasing use of mobile technology in different sectors of society to meet the needs of people on the move. In business, there is increasing use of mobile technologies for individuals to conduct their business anywhere and anytime. In healthcare, medical staff are using mobile technologies to access just-in-time information and to enter information in real time. People working in the field away from the central office use mobile technologies to access information and to communicate with other workers. Also, younger generations of learners are using mobile technologies for entertainment and socialization. These learners are using mobile devices to access information and multimedia materials and to communicate with friends. These new generations of learners do not see technology as something foreign. They readily accept technology and consider technology to be part of their lives. Moreover, the use of mobile technology is a 21st Century skill that students and workers must have to function in society.

  5. Predictors of Social Mobilization Speed

    CERN Document Server

    Alstott, Jeff; Velu, Chander

    2013-01-01

    Mobilization across social networks is becoming increasingly influential, but little is known about what traits of individuals and their relationships affect their speed of mobilization between them. We ran a global social mobilization contest and recorded personal traits of the participants and those they mobilized. We examined how those traits influenced the speed of mobilization. Individuals mobilized faster when they heard about the contest directly from the contest organization, and decreased in speed when hearing from less personal source types (e.g. family vs. media). Mobilization was faster when the mobilizer and the mobilized heard about the contest through the same source type, and slower when both individuals were in different countries. Females mobilized other females faster than males mobilized other males. Mobilization was faster with young recruiters and old recruits, and slower with old recruiters and young recruits. These findings suggest ways to optimize the speed of social mobilization.

  6. Mobile human-computer interaction perspective on mobile learning

    CSIR Research Space (South Africa)

    Botha, Adèle

    2010-10-01

    Full Text Available Applying a Mobile Human Computer Interaction (MHCI) view to the domain of education using Mobile Learning (Mlearning), the research outlines its understanding of the influences and effects of different interactions on the use of mobile technology...

  7. Wireless mobile Internet security

    CERN Document Server

    Rhee, Man Young

    2013-01-01

      The mobile industry for wireless cellular services has grown at a rapid pace over the past decade. Similarly, Internet service technology has also made dramatic growth through the World Wide Web with a wire line infrastructure. Realization for complete wired/wireless mobile Internet technologies will become the future objectives for convergence of these technologies thr

  8. Mobile Informal Learning

    NARCIS (Netherlands)

    Börner, Dirk; Glahn, Christian; Specht, Marcus

    2009-01-01

    Börner, D., Glahn, C., & Specht, M. (2009). Mobile Informal Learning. Presentation for the Education in the Wild: contextual and location-based mobile learning in action workshop at the STELLAR Alpine Rendez-Vous 2009. November, 30-December, 3, 2009, Garmisch-Partenkirchen, Germany.

  9. Intergenerational earnings mobilities

    DEFF Research Database (Denmark)

    Hussain, Azhar; Munk, Martin D.; Bonke, Jens

    2009-01-01

    comparisons of intergenerational earnings mobility. We find that intergenerational earnings mobility is found to be substantially lower when hourly wage rates rather than annual earnings are used, whether the latter are inclusive or exclusive of public transfers. Moreover, when the same specifications...

  10. Innovazione nel mobile learning

    Directory of Open Access Journals (Sweden)

    Immaculada Arnedillo-Sànchez

    2008-01-01

    Full Text Available Descrizione, da una prospettiva europea, dell’innovazione nel settore del mobile learning e l’utilizzabilita’ del mobile learning in contesti educativi. Vengono illustrate i principali progetti europei di m-learning e si esamina le prospettive pedagogiche e teoriche relative al campo.

  11. Mobile-first Bootstrap

    CERN Document Server

    Magno, Alexandre

    2013-01-01

    A practical, step-by-step tutorial on developing websites for mobile using Bootstrap.This book is for anyone who wants to get acquainted with the new features available in Bootstrap 3 and who wants to develop websites with the mobile-first feature of Bootstrap. The reader should have a basic knowledge of Bootstrap as a frontend framework.

  12. Children, Mobility, and Space

    DEFF Research Database (Denmark)

    Christensen, Pia; Romero Mikkelsen, Miguel; Nielsen, Thomas Alexander Sick

    2011-01-01

    This article discusses the potentials of a mixed methods approach to the study of children’s mobility patterns. The methodology presented here combined ethnographic fieldwork with global positioning system technology and an interactive questionnaire that children completed via mobile phone. This ...

  13. Mobile light in roses

    NARCIS (Netherlands)

    Marissen, A.; Snel, J.F.H.; Elings, A.; Warmenhoven, M.G.

    2006-01-01

    In order to quantify the claimed differences between mobile and static lighting, and to clarify the mechanisms causing these differences, an experiment with rose `First Red¿ was carried out. In a greenhouse compartment half of the area was equipped with mobile lamps, and the other half with static

  14. Mobile communication security

    NARCIS (Netherlands)

    Broek, F.M.J. van den

    2016-01-01

    Security of the mobile network Fabian van den Broek We looked at the security of the wireless connection between mobile phone and cell towers and suggested possible improvements. The security was analysed on a design level, by looking at the protocols and encryption techniques, but also on an

  15. Journalists and Mobilizing Information

    Science.gov (United States)

    Lemert, James B.; And Others

    1977-01-01

    Analysis of the items in 11 daily newspapers revealed that approximately 36 percent included some "mobilizing information"--information about ways readers might express their preferences and desires. However, only 6 percent of the items containing mobilizing information dealt with political issues or the political process. (GW)

  16. Mastering Mobile Security

    Science.gov (United States)

    Panettieri, Joseph C.

    2007-01-01

    Without proper security, mobile devices are easy targets for worms, viruses, and so-called robot ("bot") networks. Hackers increasingly use bot networks to launch massive attacks against eCommerce websites--potentially targeting one's online tuition payment or fundraising/financial development systems. How can one defend his mobile systems against…

  17. Designing for mobile interaction

    DEFF Research Database (Denmark)

    Nazzi, Elena

    2009-01-01

    The theme of this PhD project is designing for mobile interaction with devices and services, for the accessing, making, and sharing of information, taking into account the dynamic physical and social settings that embrace this interaction. To narrow down this theme, the whole project focuses...... on the exploitation of social interaction --- in particular among senior citizens --- to enhance and support mobile interaction....

  18. Mobile systems development

    DEFF Research Database (Denmark)

    Hosbond, Jens Henrik

    2005-01-01

    This paper takes a systems development perspective on mobility, building on preliminary findings of an on-going multiple case study covering 7 companies. The questions driving this paper are: What are the challenges facing development practice in the mobile industry, how do they affect practice...

  19. Mobile Christian - shuttle flight

    Science.gov (United States)

    2009-01-01

    Erin Whittle, 14, (seated) and Brianna Johnson, 14, look on as Louis Stork, 13, attempts a simulated landing of a space shuttle at StenniSphere. The young people were part of a group from Mobile Christian School in Mobile, Ala., that visited StenniSphere on April 21.

  20. Mobile Apps for Librarians

    Science.gov (United States)

    Power, June L.

    2013-01-01

    In an increasing mobile environment, library and reading-related activities often take place on a phone or tablet device. Not only does this mean that library Web sites must keep mobile navigability in mind, but also develop and utilize apps that allow patrons to interact with information and with libraries. While apps do not serve every purpose,…

  1. Mobile Media Photography

    DEFF Research Database (Denmark)

    Svabo, Connie; Shanks, Michael

    2013-01-01

    Mobile media photography marks a shift in orientation from the image towards photography as a mode of engagement. This leads is to explore the processes of experience and documentation that mobile media help to constitute. We unfold two aspects of the process of photography: photography as temporal...

  2. Mobile4D platform

    CSIR Research Space (South Africa)

    Botha, Adèle

    2010-05-01

    Full Text Available The mobile telecommunications story in Africa and the developing world is a remarkable one. Africa's mobile cellular growth rate has been the highest of any region over the past 5 years, averaging close to 60% year on year. There have been large...

  3. Children, mobility, and space

    DEFF Research Database (Denmark)

    Christensen, Pia; Mikkelsen, Miguel Romero; Nielsen, Thomas Alexander Sick

    2011-01-01

    This article discusses the potentials of a mixed methods approach to the study of children’s mobility patterns. The methodology presented here combined ethnographic fieldwork with global positioning system technology and an interactive questionnaire that children completed via mobile phone. This ...

  4. Mobile Learning Anytime, Anywhere

    Science.gov (United States)

    Hlodan, Oksana

    2010-01-01

    Some educational institutions are taking the leap to mobile learning (m-learning) by giving out free iPods. For example, Abilene Christian University gave iPods or iPhones to freshman students and developed 15 Web applications specifically for the mobile devices. The iPod is not the only ubiquitous m-learning device. Any technology that connects…

  5. Intergenerational earnings mobilities

    DEFF Research Database (Denmark)

    Hussain, Azhar; Munk, Martin D.; Bonke, Jens

    2009-01-01

    This article gives various estimates of intergenerational earnings mobility by applying different earning periods, age brackets, and earning components. The methodology enables us to investigate how sensitive results are to different delimitations and, thereby, to make more accurate international...... comparisons of intergenerational earnings mobility. We find that intergenerational earnings mobility is found to be substantially lower when hourly wage rates rather than annual earnings are used, whether the latter are inclusive or exclusive of public transfers. Moreover, when the same specifications...... are applied for Denmark as for other countries, we find that intergenerational earnings mobility from father to son in Denmark is on the same level as in Sweden, Norway, and Finland, whereas the intergenerational earnings mobility in all the Nordic countries is found to be higher than in the United Kingdom...

  6. Contextualizing mobile IT

    DEFF Research Database (Denmark)

    Messeter, Jörn; Brandt, Eva; Halse, Joachim

    2004-01-01

    Information and communication technologies are moving into the era of ubiquitous computing, with increased density of technology and increased mobility and continuity in use. From a design perspective, addressing the accommodation and coordination of multiple devices and services in situated use...... across different contexts is becoming increasingly important. In the COMIT project, ethnographic fieldwork has been combined with participatory design engaging users, designers and researchers in order to explore mobile IT use as well as the design of mobile IT concepts. Four seclected scenarios from...... the project are presented and discussed regarding implications for the design of mobile IT devices, with particular focus on (1) coping with multiple social contexts, and (2) the configuration and connectivity of mobile devices...

  7. Mobilities, Futures & the City

    DEFF Research Database (Denmark)

    Freudendal-Pedersen, Malene; Kesselring, Sven

    2016-01-01

    The future of cities and regions will be strongly shaped by the mobilities of people, goods, modes of transport, waste and information. In many ways, the ‘why and ‘for what’ often get lost in discourses on planning and designing mobilities. The predominant planning paradigm still conceptualizes...... the future of cities and mobilities as a matter of rather more efficient technologies than of social cohesion, integration and connectivity. Sustainable mobility needs the mobilities of ideas and concepts and the reflexivity of policies. Communicative planning theory and the ‘argumentative turn’ have given...... this article, was to explicitly provide an intersection for reflexivity, interdisciplinarity and exchange, to foster the creation of such stories....

  8. Mobilities and Representations

    DEFF Research Database (Denmark)

    Thelle, Mikkel

    2017-01-01

    , literature, and film. Moreover, we hope the authors of future reviews will reflect on the ways they approached those representations. Such commentaries would provide valuable methodological insights, and we hope to begin that effort with this interview. We have asked four prominent mobility scholars......As the centerpiece of the eighth T2M yearbook, the following interview about representations of mobility signals a new and exciting focus area for Mobility in History. In future issues we hope to include reviews that grapple more with how mobilities have been imagined and represented in the arts...... to consider how they and their peers are currently confronting representations of mobility. This is particularly timely given the growing academic focus on practices, material mediation, and nonrepresentational theories, as well as on bodily reactions, emotions, and feelings that, according to those theories...

  9. Mobile Collocated Interactions

    DEFF Research Database (Denmark)

    Lucero, Andrés; Clawson, James; Lyons, Kent

    2015-01-01

    Mobile devices such as smartphones and tablets were originally conceived and have traditionally been utilized for individual use. Research on mobile collocated interactions has been looking at situations in which collocated users engage in collaborative activities using their mobile devices, thus...... going from personal/individual toward shared/multiuser experiences and interactions. However, computers are getting smaller, more powerful, and closer to our bodies. Therefore, mobile collocated interactions research, which originally looked at smartphones and tablets, will inevitably include ever......-smaller computers, ones that can be worn on our wrists or other parts of the body. The focus of this workshop is to bring together a community of researchers, designers and practitioners to explore the potential of extending mobile collocated interactions to the use of wearable devices....

  10. Mobile Health Information System: A Mobile App

    African Journals Online (AJOL)

    Steve

    It also serves as an inventory control where statistics about a particular illness is known. Generally, Mobile Health Information System (MHIS) is built to help manage, control, and store health information, and at the same time help the health workers on field work to pass accurate and timely report to the necessary quarters.

  11. Tactical Mobile Communications (Communications tactiques mobiles)

    Science.gov (United States)

    1999-11-01

    Stations that are not willing to transmit may10 switch to a sleeping mode. To enable mobile stations to be 8 . addressed and reached even when sleeping ...probe - 31 symbols of a repeated 16 symbol Frank-Heimiller polyphase code. Regularly Re-inserted Preamble - 103 symbols Figure 5 - Waveform structure. A

  12. Gone Mobile? (Mobile Libraries Survey 2010)

    Science.gov (United States)

    Thomas, Lisa Carlucci

    2010-01-01

    Librarians, like patrons and researchers, are caught between traditional library service models and the promise of evolving information technologies. In recent years, professional conferences have strategically featured programs and presentations geared toward building a mobile agenda and adapting or adopting services to meet new demands of mobile…

  13. Friend of GATA (FOG interacts with the nucleosome remodeling and deacetylase complex (NuRD to support primitive erythropoiesis in Xenopus laevis.

    Directory of Open Access Journals (Sweden)

    Mizuho S Mimoto

    Full Text Available Friend of GATA (FOG plays many diverse roles in adult and embryonic hematopoiesis, however the mechanisms by which it functions and the roles of potential interaction partners are not completely understood. Previous work has shown that overexpression of FOG in Xenopus laevis causes loss of blood suggesting that in contrast to its role in mammals, FOG might normally function to repress erythropoiesis in this species. Using loss-of-function analysis, we demonstrate that FOG is essential to support primitive red blood cell (RBC development in Xenopus. Moreover, we show that it is specifically required to prevent excess apoptosis of circulating primitive RBCs and that in the absence of FOG, the pro-apoptotic gene Bim-1 is strongly upregulated. To identify domains of FOG that are essential for blood development and, conversely, to begin to understand the mechanism by which overexpressed FOG represses primitive erythropoiesis, we asked whether FOG mutants that are unable to interact with known co-factors retain their ability to rescue blood formation in FOG morphants and whether they repress erythropoiesis when overexpressed in wild type embryos. We find that interaction of FOG with the Nucleosome Remodeling and Deacetylase complex (NuRD, but not with C-terminal Binding Protein, is essential for normal primitive RBC development. In contrast, overexpression of all mutant and wild type constructs causes a comparable repression of primitive erythropoiesis. Together, our data suggest that a requirement for FOG and its interaction with NuRD during primitive erythropoiesis are conserved in Xenopus and that loss of blood upon FOG overexpression is due to a dominant-interfering effect.

  14. What is mobile in entrepreneurship?

    OpenAIRE

    Lundmark, Erik; Waern, Åsa

    2008-01-01

    This report focuses on the relation between entrepreneurship and different aspects of mobility and provides a basis for further studies. To date the entrepreneurship field has focused little on the role of mobility and the studies that do, have predominantly focused on labour mobility. We argue that entrepreneurship and mobility are intertwined and that limiting the concept of mobility to labour mobility limits our view of the entrepreneurial process. The study is based on a literature review...

  15. HUMAN INTERACTION WITH MOBILE APPLICATIONS

    OpenAIRE

    Alin Zamfiroiu; Emanuel Herteliu; Bogdan Vintila

    2012-01-01

    Computing - human interaction is a very important paradigm because informatics applications are created to be used by people via human interaction. Nowadays mobile applications are more used so is necessarily to talk about mobile - human interaction. In this paper types of mobile devices are presented. Citizen oriented character of mobile application and his utility are described. Different means of interactions with mobile devices are analyzed and in the end of the paper direction of mobile ...

  16. Client Mobile Software Design Principles for Mobile Learning Systems

    Directory of Open Access Journals (Sweden)

    Qing Tan

    2009-01-01

    Full Text Available In a client-server mobile learning system, client mobile software must run on the mobile phone to acquire, package, and send student’s interaction data via the mobile communications network to the connected mobile application server. The server will receive and process the client data in order to offer appropriate content and learning activities. To develop the mobile learning systems there are a number of very important issues that must be addressed. Mobile phones have scarce computing resources. They consist of heterogeneous devices and use various mobile operating systems, they have limitations with their user/device interaction capabilities, high data communications cost, and must provide for device mobility and portability. In this paper we propose five principles for designing Client mobile learning software. A location-based adaptive mobile learning system is presented as a proof of concept to demonstrate the applicability of these design principles.

  17. Mobile sensing systems.

    Science.gov (United States)

    Macias, Elsa; Suarez, Alvaro; Lloret, Jaime

    2013-12-16

    Rich-sensor smart phones have made possible the recent birth of the mobile sensing research area as part of ubiquitous sensing which integrates other areas such as wireless sensor networks and web sensing. There are several types of mobile sensing: individual, participatory, opportunistic, crowd, social, etc. The object of sensing can be people-centered or environment-centered. The sensing domain can be home, urban, vehicular… Currently there are barriers that limit the social acceptance of mobile sensing systems. Examples of social barriers are privacy concerns, restrictive laws in some countries and the absence of economic incentives that might encourage people to participate in a sensing campaign. Several technical barriers are phone energy savings and the variety of sensors and software for their management. Some existing surveys partially tackle the topic of mobile sensing systems. Published papers theoretically or partially solve the above barriers. We complete the above surveys with new works, review the barriers of mobile sensing systems and propose some ideas for efficiently implementing sensing, fusion, learning, security, privacy and energy saving for any type of mobile sensing system, and propose several realistic research challenges. The main objective is to reduce the learning curve in mobile sensing systems where the complexity is very high.

  18. Limits of social mobilization.

    Science.gov (United States)

    Rutherford, Alex; Cebrian, Manuel; Dsouza, Sohan; Moro, Esteban; Pentland, Alex; Rahwan, Iyad

    2013-04-16

    The Internet and social media have enabled the mobilization of large crowds to achieve time-critical feats, ranging from mapping crises in real time, to organizing mass rallies, to conducting search-and-rescue operations over large geographies. Despite significant success, selection bias may lead to inflated expectations of the efficacy of social mobilization for these tasks. What are the limits of social mobilization, and how reliable is it in operating at these limits? We build on recent results on the spatiotemporal structure of social and information networks to elucidate the constraints they pose on social mobilization. We use the DARPA Network Challenge as our working scenario, in which social media were used to locate 10 balloons across the United States. We conduct high-resolution simulations for referral-based crowdsourcing and obtain a statistical characterization of the population recruited, geography covered, and time to completion. Our results demonstrate that the outcome is plausible without the presence of mass media but lies at the limit of what time-critical social mobilization can achieve. Success relies critically on highly connected individuals willing to mobilize people in distant locations, overcoming the local trapping of diffusion in highly dense areas. However, even under these highly favorable conditions, the risk of unsuccessful search remains significant. These findings have implications for the design of better incentive schemes for social mobilization. They also call for caution in estimating the reliability of this capability.

  19. Mobile Sensing Systems

    Directory of Open Access Journals (Sweden)

    Elsa Macias

    2013-12-01

    Full Text Available Rich-sensor smart phones have made possible the recent birth of the mobile sensing research area as part of ubiquitous sensing which integrates other areas such as wireless sensor networks and web sensing. There are several types of mobile sensing: individual, participatory, opportunistic, crowd, social, etc. The object of sensing can be people-centered or environment-centered. The sensing domain can be home, urban, vehicular… Currently there are barriers that limit the social acceptance of mobile sensing systems. Examples of social barriers are privacy concerns, restrictive laws in some countries and the absence of economic incentives that might encourage people to participate in a sensing campaign. Several technical barriers are phone energy savings and the variety of sensors and software for their management. Some existing surveys partially tackle the topic of mobile sensing systems. Published papers theoretically or partially solve the above barriers. We complete the above surveys with new works, review the barriers of mobile sensing systems and propose some ideas for efficiently implementing sensing, fusion, learning, security, privacy and energy saving for any type of mobile sensing system, and propose several realistic research challenges. The main objective is to reduce the learning curve in mobile sensing systems where the complexity is very high.

  20. Mobile Sensing Systems

    Science.gov (United States)

    Macias, Elsa; Suarez, Alvaro; Lloret, Jaime

    2013-01-01

    Rich-sensor smart phones have made possible the recent birth of the mobile sensing research area as part of ubiquitous sensing which integrates other areas such as wireless sensor networks and web sensing. There are several types of mobile sensing: individual, participatory, opportunistic, crowd, social, etc. The object of sensing can be people-centered or environment-centered. The sensing domain can be home, urban, vehicular… Currently there are barriers that limit the social acceptance of mobile sensing systems. Examples of social barriers are privacy concerns, restrictive laws in some countries and the absence of economic incentives that might encourage people to participate in a sensing campaign. Several technical barriers are phone energy savings and the variety of sensors and software for their management. Some existing surveys partially tackle the topic of mobile sensing systems. Published papers theoretically or partially solve the above barriers. We complete the above surveys with new works, review the barriers of mobile sensing systems and propose some ideas for efficiently implementing sensing, fusion, learning, security, privacy and energy saving for any type of mobile sensing system, and propose several realistic research challenges. The main objective is to reduce the learning curve in mobile sensing systems where the complexity is very high. PMID:24351637

  1. Mobile and contextual learning

    Directory of Open Access Journals (Sweden)

    Agnes Kukulska-Hulme

    2009-12-01

    Full Text Available Is mobile learning just a part of everyday learning? This is a relevant question in an age when most people throughout the world now have access to mobile phones and mobility is increasingly taken for granted. In one sense, mobile learning is no different to carrying a textbook or learning through conversations at home, as part of formal education or in the workplace. The technology may be more engaging, but is the learning any different? Despite the ubiquity of mobile phones, smartphones, mp3 players and, increasingly, access to Wi-Fi connections and GPS navigation, the reasons for using mobile and wireless technologies in education are not yet widely known. There is significant specialist expertise, built from ten years of research (and more recently, reflective practice to demonstrate the unique characteristics of mobile learning, which include orchestrating shared learning with personal devices across formal and informal settings, providing immediately useful information, offering timely revision and reflection, connecting real and virtual locations, and enriching field trips and cultural visits. This knowledge needs to be disseminated and examined from new perspectives.

  2. Cooperative Mobile Web Browsing

    Directory of Open Access Journals (Sweden)

    Zhang Q

    2009-01-01

    Full Text Available This paper advocates a novel approach for mobile web browsing based on cooperation among wireless devices within close proximity operating in a cellular environment. In the actual state of the art, mobile phones can access the web using different cellular technologies. However, the supported data rates are not sufficient to cope with the ever increasing traffic requirements resulting from advanced and rich content services. Extending the state of the art, higher data rates can only be achieved by increasing complexity, cost, and energy consumption of mobile phones. In contrast to the linear extension of current technology, we propose a novel architecture where mobile phones are grouped together in clusters, using a short-range communication such as Bluetooth, sharing, and accumulating their cellular capacity. The accumulated data rate resulting from collaborative interactions over short-range links can then be used for cooperative mobile web browsing. By implementing the cooperative web browsing on commercial mobile phones, it will be shown that better performance is achieved in terms of increased data rate and therefore reduced access times, resulting in a significantly enhanced web browsing user experience on mobile phones.

  3. Editorial: Mobile (March 2010

    Directory of Open Access Journals (Sweden)

    Thomas Kunz

    2010-02-01

    Full Text Available Open source software and hardware has become an accepted way of developing new and interesting applications in many information and communication technology domains: operating systems, databases, Web infrastructure, and applications. It's not surprising that with the increasing popularity of mobile handheld devices, users and researchers have explored the power of open approaches to providing innovative new applications and services in this domain. However, unlike personal computers and the Internet, mobile handsets were tightly controlled by mobile network operators (MNOs who developed a vertical ecosystem by integrating the communication infrastructure, the handheld device hardware, and often the applications installed on those devices. The software and protocols running the mobile communications infrastructure and devices are often standardized by membership-only bodies, where large MNOs and manufacturers have a predominant influence. These players invest significant financial resources into shaping the industry along their vision to gain a competitive advantage. A current example is the ongoing battle about the dominant radio access technology for 4G cellular systems: LTE vs. Wimax. These trends have changed recently. Companies such as Google, Nokia, or Openmoko and Industry Alliances such as the Open Handset Alliance are providing the core building blocks, both in hardware as well as software, of increasingly open mobile devices. This issue of the OSBR reviews the relevant trends in the open mobile platform space from a number of perspectives. As the articles in these issue show, there is a lot of exciting ongoing work that brings the power of open source development to the mobile space. This trend is not just confined to the mobile devices as there are also efforts in the development of open mobile infrastructure elements and whole systems.

  4. Mobile Context Toolbox

    DEFF Research Database (Denmark)

    Stopczynski, Arkadiusz; Larsen, Jakob Eg; Skomail, Lukasz

    2010-01-01

    In this paper we describe an open framework utilizing sensors and application data on the Maemo mobile platform enabling rapid prototyping of context-aware mobile applications. The framework has an extensible layered architecture allowing new hardware and software sensors and features to be added...... to the context framework. We present initial results from in-the-wild experiments where contextual data was acquired using the tool. In the experiments 6 participants were using a Nokia N900 mobile phone continuously with a logger application for an average of 33 days. The study has provided valuable insights...

  5. Mobile Game Probes

    DEFF Research Database (Denmark)

    Borup Lynggaard, Aviaja

    2006-01-01

    This paper will examine how probes can be useful for game designers in the preliminary phases of a design process. The work is based upon a case study concerning pervasive mobile phone games where Mobile Game Probes have emerged from the project. The new probes are aimed towards a specific target...... group and the goal is to specify the probes so they will cover the most relevant areas for our project. The Mobile Game Probes generated many interesting results and new issues occurred, since the probes came to be dynamic and favorable for the process in new ways....

  6. Mobile sociology. 2000.

    Science.gov (United States)

    Urry, John

    2010-01-01

    This article seeks to develop a manifesto for a sociology concerned with the diverse mobilities of peoples, objects, images, information, and wastes; and of the complex interdependencies between, and social consequences of, such diverse mobilities. A number of key concepts relevant for such a sociology are elaborated: 'gamekeeping', networks, fluids, scapes, flows, complexity and iteration. The article concludes by suggesting that a 'global civil society' might constitute the social base of a sociology of mobilities as we move into the twenty-first century.

  7. MOBILIZING KNOWLEDGE IN DENMARK

    DEFF Research Database (Denmark)

    Holm, Claus

    2011-01-01

    competition state – that is to be realized by fulfilling the ambitions for comprehensive knowledge mobilization through education.1 On the other hand, Denmark is having a hard time finding a broad consensus when what is, in principle, supposed to be a common political will is to be translated into knowledge...... mobilization. This lack of consensus can be traced to the political-administrative level and to the relationship between representatives of education research and representatives of the education system. So, if the concept of knowledge mobilization is not especially exploited in Denmark, it can be seen...

  8. Mobile ad hoc networking

    CERN Document Server

    John Wiley & Sons

    2004-01-01

    "Assimilating the most up-to-date information on research and development activities in this rapidly growing area, Mobile Ad Hoc Networking covers physical, data link, network, and transport layers, as well as application, security, simulation, and power management issues in sensor, local area, personal, and mobile ad hoc networks. Each of the book's sixteen chapters has been written by a top expert and discusses in-depth the most important topics in the field. Mobile Ad Hoc Networking is an excellent reference and guide for professionals seeking an in-depth examination of topics that also provides a comprehensive overview of the current state-of-the-art."--Jacket.

  9. Social Perspectives on Mobility

    DEFF Research Database (Denmark)

    Uth Thomsen, Thyra; Gudmundsson, Henrik; Drewes Nielsen, Lise

    Globalisation is heavily dependent on physical transport, as people and goods travel over longer distances and with higher frequency. Movement and mobility have become integrated parts of late modern identity and practice, and a state of flux can be sensed everywhere. Bringing together the latest...... interdisciplinary theoretical approaches with empirical case studies analysing and appraising innovative policies from Scandinavia, this volume demonstrates that mobility research is a key issue within social enquiry. It addresses three broad themes. Firstly, mobility as a constructed social reality, examining how...

  10. Mobile communication and intermediality

    DEFF Research Database (Denmark)

    Helles, Rasmus

    2013-01-01

    The article argues the importance of intermediality as a concept for research in mobile communication and media. The constant availability of several, partially overlapping channels for communication (texting, calls, email, Facebook, etc.) requires that we adopt an integrated view of the various...... communicative affordances of mobile devices in order to understand how people choose between them for different purposes. It is argued that mobile communication makes intermediality especially central, as the choice of medium is detached from the location of stationary media and begins to follow the user across...

  11. Mobile computing for radiology.

    Science.gov (United States)

    Auffermann, William F; Chetlen, Alison L; Sharma, Arjun; Colucci, Andrew T; DeQuesada, Ivan M; Grajo, Joseph R; Kung, Justin W; Loehfelm, Thomas W; Sherry, Steven J

    2013-12-01

    The rapid advances in mobile computing technology have the potential to change the way radiology and medicine as a whole are practiced. Several mobile computing advances have not yet found application to the practice of radiology, while others have already been applied to radiology but are not in widespread clinical use. This review addresses several areas where radiology and medicine in general may benefit from adoption of the latest mobile computing technologies and speculates on potential future applications. Copyright © 2013 AUR. Published by Elsevier Inc. All rights reserved.

  12. MOBILE GAME HALMA MULTIPLAYER

    Directory of Open Access Journals (Sweden)

    Muhammad Dhimas

    2015-04-01

    Full Text Available Handphone besides as a communication tool also has a function as a medium of entertainment. Various multimedia services and communications facilities contained therein, one Bluetooth. Games is one application that always exist in the mobile phone, and with a wider variety of games development by utilizing the services in mobile. In this research, the development of the multiplayer games for mobile phones utilizing Bluetooth communication media using the programming language Java Micro Edition (J2ME. Design method using the grapple, NetBeans IDE 6.1 is used as tools to assist programming.

  13. Data mining mobile devices

    CERN Document Server

    Mena, Jesus

    2013-01-01

    With today's consumers spending more time on their mobiles than on their PCs, new methods of empirical stochastic modeling have emerged that can provide marketers with detailed information about the products, content, and services their customers desire.Data Mining Mobile Devices defines the collection of machine-sensed environmental data pertaining to human social behavior. It explains how the integration of data mining and machine learning can enable the modeling of conversation context, proximity sensing, and geospatial location throughout large communities of mobile users

  14. Intergenerational earnings mobilities

    DEFF Research Database (Denmark)

    Hussain, Azhar; Munk, Martin D.; Bonke, Jens

    2009-01-01

    This article gives various estimates of intergenerational earnings mobility by applying different earning periods, age brackets, and earning components. The methodology enables us to investigate how sensitive results are to different delimitations and, thereby, to make more accurate international...... are applied for Denmark as for other countries, we find that intergenerational earnings mobility from father to son in Denmark is on the same level as in Sweden, Norway, and Finland, whereas the intergenerational earnings mobility in all the Nordic countries is found to be higher than in the United Kingdom...

  15. Mobility Management in Denmark

    DEFF Research Database (Denmark)

    Guasco, Clement Nicolas

    2012-01-01

    There is today an undisputed superiority of cars on any other means of transportation, both practically and psychologically. However, the reliance on cars as the main means of transportation has led to a number of social and environmental problems. Mobility Management aims at managing the way...... people fulfil their mobility needs so that they rely less on their cars and start using alternatives such as bicycles and public transport. This article analyses the possibility to use businesses as vectors for introducing Mobility Management within social practices in Denmark in the light of the recent...

  16. Intergenerational earnings mobilities

    DEFF Research Database (Denmark)

    Hussain, Azhar; Munk, Martin D.; Bonke, Jens

    2009-01-01

    This article gives various estimates of intergenerational earnings mobility by applying different earning periods, age brackets, and earning components. The methodology enables us to investigate how sensitive results are to different delimitations and, thereby, to make more accurate international...... and the United States....... are applied for Denmark as for other countries, we find that intergenerational earnings mobility from father to son in Denmark is on the same level as in Sweden, Norway, and Finland, whereas the intergenerational earnings mobility in all the Nordic countries is found to be higher than in the United Kingdom...

  17. Mobile networks architecture

    CERN Document Server

    Perez, Andre

    2013-01-01

    This book explains the evolutions of architecture for mobiles and summarizes the different technologies:- 2G: the GSM (Global System for Mobile) network, the GPRS (General Packet Radio Service) network and the EDGE (Enhanced Data for Global Evolution) evolution;- 3G: the UMTS (Universal Mobile Telecommunications System) network and the HSPA (High Speed Packet Access) evolutions:- HSDPA (High Speed Downlink Packet Access),- HSUPA (High Speed Uplink Packet Access),- HSPA+;- 4G: the EPS (Evolved Packet System) network.The telephone service and data transmission are the

  18. Mobile Internet: Politics of Mobile Code and Networks

    NARCIS (Netherlands)

    L. Mosemghvdlishvili (Lela)

    2015-01-01

    markdownabstract__Abstract__ The term mobile internet refers to accessing the internet through (cellular) mobile devices and represents the convergence of mobile telephony, internet services, and personal computing. Whereas iMode (mobile internet and data services) was introduced in Japan as long

  19. Proxemic Mobile Collocated Interactions

    DEFF Research Database (Denmark)

    Porcheron, Martin; Lucero, Andrés; Quigley, Aaron

    2016-01-01

    Recent research on mobile collocated interactions has been looking at situations in which collocated users engage in collaborative activities using their mobile devices. However, existing practices fail to fully account for the culturally-dependent spatial relationships between people and their d......Recent research on mobile collocated interactions has been looking at situations in which collocated users engage in collaborative activities using their mobile devices. However, existing practices fail to fully account for the culturally-dependent spatial relationships between people...... would allow devices to not only react to presence and interaction, but also other indicators, such as the interpersonal distance people naturally use in everyday life. The aim of this one-day workshop is to bring together a community of researchers, designers and practitioners who are interested...

  20. Promoting regional mobility

    DEFF Research Database (Denmark)

    Jensen, Anne

    Pricing of transport has been part of EU's common transport policy since this gained momentum in the early 1990s. Since then, it has been closely connected to the trans-European transport network (TEN-T) and to rising demands of efficient mobility systems at a local, regional and Community scale....... Development of pricing policies is contested at Community level and has taken place in a clash between different policy rationalities. Significantly though, the effects of the pricing policies are closely related to regional mobility systems, e.g. through financing large trans-border infrastructure projects...... and establishing common technical charging systems thus changing the conditions for regional mobility. This paper explores how policies of infrastructure pricing shape new ways of governing mobility which influences trans-border, regional policy-making. The key findings are that there is a tendency to include...

  1. Towards urban mobility designs

    DEFF Research Database (Denmark)

    Lanng, Ditte Bendix; Harder, Henrik; Jensen, Ole B.

    2012-01-01

    Functionalist traffic design aims at a rational organisation of movement from A to B in a segregated and uniform physical environment. Such urban areas have been criticized for being exclusively functional and lacking sensuous and social qualities. Recent research on mobilities challenges the more...... are concerned with. - Third, we introduce an embodied mobile perspective through insights from the mobilities turn. - Fourth, through our own journey experience en route we present a mapping of the transit site. - We conclude by outlining a few propositions on the design challenge and design potentials......, in relation to re-conceptualisation of the transit site, to mapping, and to design intervention. These propositions form part of our on-going work with urban mobility designs....

  2. Intergenerational earnings mobilities

    DEFF Research Database (Denmark)

    Hussain, Azhar; Munk, Martin D.; Bonke, Jens

    2009-01-01

    This article gives various estimates of intergenerational earnings mobility by applying different earning periods, age brackets, and earning components. The methodology enables us to investigate how sensitive results are to different delimitations and, thereby, to make more accurate international...

  3. Mobile Learning Applications Audit

    Directory of Open Access Journals (Sweden)

    Paul POCATILU

    2010-01-01

    Full Text Available While mobile learning (m-learning applications have proven their value in educational activities, there is a need to measure their reliability, accessibility and further more their trustworthiness. Mobile devices are far more vulnerable then classic computers and present inconvenient interfaces due to their size, hardware limitations and their mobile connectivity. Mobile learning applications should be audited to determine if they should be trusted or not, while multimedia contents like automatic speech recognition (ASR can improve their accessibility. This article will start with a brief introduction on m-learning applications, then it will present the audit process for m-learning applications, it will iterate their specific security threats, it will define the ASR process, and it will elaborate how ASR can enhance accessibility of these types of applications.

  4. Cooperative Mobile Web Browsing

    DEFF Research Database (Denmark)

    Perrucci, GP; Fitzek, FHP; Zhang, Qi

    2009-01-01

    extension of current technology, we propose a novel architecture where mobile phones are grouped together in clusters, using a short-range communication such as Bluetooth, sharing, and accumulating their cellular capacity. The accumulated data rate resulting from collaborative interactions over short......This paper advocates a novel approach for mobile web browsing based on cooperation among wireless devices within close proximity operating in a cellular environment. In the actual state of the art, mobile phones can access the web using different cellular technologies. However, the supported data...... rates are not sufficient to cope with the ever increasing trafic requirements resulting from advanced and rich content services. Extending the state of the art, higher data rates can only be achieved by increasing complexity, cost, and energy consumption of mobile phones. In contrast to the linear...

  5. jQuery Mobile

    CERN Document Server

    Reid, Jon

    2011-01-01

    Native apps have distinct advantages, but the future belongs to mobile web apps that function on a broad range of smartphones and tablets. Get started with jQuery Mobile, the touch-optimized framework for creating apps that look and behave consistently across many devices. This concise book provides HTML5, CSS3, and JavaScript code examples, screen shots, and step-by-step guidance to help you build a complete working app with jQuery Mobile. If you're already familiar with the jQuery JavaScript library, you can use your existing skills to build cross-platform mobile web apps right now. This b

  6. Mobility Data Analytics Center.

    Science.gov (United States)

    2016-01-01

    Mobility Data Analytics Center aims at building a centralized data engine to efficiently manipulate : large-scale data for smart decision making. Integrating and learning the massive data are the key to : the data engine. The ultimate goal of underst...

  7. Persuasive Mobile Health Applications

    Science.gov (United States)

    Garcia Wylie, Carlos; Coulton, Paul

    With many industrialized societies bearing the cost of an increasingly sedentary lifestyle on the health of their populations there is a need to find new ways of encouraging physical activity to promote better health and well being. With the increasing power of mobile phones and the recent emergence of personal heart rate monitors, aimed at dedicated amateur runners, there is now a possibility to develop “Persuasive Mobile Health Applications” to promote well being through the use of real-time physiological data and persuade users to adopt a healthier lifestyle. In this paper we present a novel general health monitoring software for mobile phones called Heart Angel. This software is aimed at helping users monitor, record, as well as improve their fitness level through built-in cardio-respiratory tests, a location tracking application for analyzing heart rate exertion over time and location, and a fun mobile-exergame called Health Defender.

  8. Mens mobile health

    DEFF Research Database (Denmark)

    Levisen, Vinie Diana Hvidbak; Castaño, Francisco Mansilla; Jensen, Camilla Skovbjerg

    2016-01-01

    Preferences Keywords: Men with little or no education, physical health, mobile health application. Types of presentations: First presentation preference: E-Poster presentations Second presentation preference: Pitch presentation Abstract Background: Men mobile health contributes knowledge of how mobile health...... applications affect the physical activity levels by men with little or no education and the frequency of how often they think and do something to promote their health. Men with little or no education have both the lowest life expectancy and longest patient delay, and there are not conducted researches...... and control group. N = 71, 20-62 years old men, little or no education, employed at industrial company. Intervention group N= 35, control group N = 36. There is performed a baseline, an 6 months intervention period, outcome measurement. The intervention: Intervention group use mobile application, registration...

  9. Making Everyday Mobility

    DEFF Research Database (Denmark)

    Wind, Simon

    practices. Specific heterogeneous configurations in mobility practices facilitate instrumental movement of family members, but can also engender care, quality time of togetherness, recreational and productive in-betweens, as well as sensorial and emotional experiences through the orchestration of affective...

  10. Mobile Tools | Smokefree 60+

    Science.gov (United States)

    These mobile resources can help you quit when you're on the go. SmokefreeTXT SmokefreeTXT is a mobile text messaging service designed for adults across the United States who are trying to quit smoking. The program offers 24/7 encouragement, advice, and tips to help smokers quit smoking and stay quit. If you are interested in signing up, fill out this form.

  11. REST based mobile applications

    Science.gov (United States)

    Rambow, Mark; Preuss, Thomas; Berdux, Jörg; Conrad, Marc

    2008-02-01

    Simplicity is the major advantage of REST based webservices. Whereas SOAP is widespread in complex, security sensitive business-to-business aplications, REST is widely used for mashups and end-user centric applicatons. In that context we give an overview of REST and compare it to SOAP. Furthermore we apply the GeoDrawing application as an example for REST based mobile applications and emphasize on pros and cons for the use of REST in mobile application scenarios.

  12. Soft Soil Mobility

    Science.gov (United States)

    2016-02-11

    any other provision of law , no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a...TYPE Final 3. DATES COVERED (From - To) 4. TITLE AND SUBTITLE Test Operations Procedure (TOP) 02-2-619A Soft Soil Mobility 5a. CONTRACT NUMBER...TOP supersedes TOP 02-2-619, Soft Soil Vehicle Mobility, dated 21 May 1970. Marginal notations are not used in this revision to identify changes

  13. Emerging Mobile Networking Architectures

    Science.gov (United States)

    2006-12-01

    years to address this problem area. Basic concepts began as early a the 1970s with DARPA sponsored "packet radio" research, yet a new flurry of design...routing, MAC , middleware). The appropriate mechanisms are also not orthogonal to the intended application or deployment scenario of the mobile ad hoc...technology that can improve the capability of future mobile architectures Original work dates back to DARPA packet radio work 1970s More recent flurry of

  14. Mobile multiple access study

    Science.gov (United States)

    1977-01-01

    Multiple access techniques (FDMA, CDMA, TDMA) for the mobile user and attempts to identify the current best technique are discussed. Traffic loading is considered as well as voice and data modulation and spacecraft and system design. Emphasis is placed on developing mobile terminal cost estimates for the selected design. In addition, design examples are presented for the alternative techniques of multiple access in order to compare with the selected technique.

  15. "Mobile Fashion" Application

    OpenAIRE

    Kashanipour, Morvarid

    2012-01-01

    This master thesis investigates studies on fashion oriented people according to the "Outfit-Centric Accessories" concept. The outfit-centric accessories concept originated from recent research study by Juhlin and Zhang (2011) about mobile phone representation in fashion and Aesthetic of Interaction area of Human-Computer Interaction (HCI). The term outfit-centric accessories originated from clothing and wearer. In this concept an outfit is playing a role as the centerpiece and a mobile phone ...

  16. Mobile Intelligent Autonomous Systems

    OpenAIRE

    Jitendra R. Raol; Ajith Gopal

    2010-01-01

    Mobile intelligent autonomous systems (MIAS) is a fast emerging research area. Although it can be regarded as a general R&D area, it is mainly directed towards robotics. Several important subtopics within MIAS research are:(i) perception and reasoning, (ii) mobility and navigation,(iii) haptics and teleoperation, (iv) image fusion/computervision, (v) modelling of manipulators, (vi) hardware/software architectures for planning and behaviour learning leadingto robotic architecture, (vii) ve...

  17. Social Perspectives on Mobility

    DEFF Research Database (Denmark)

    Uth Thomsen, Thyra; Gudmundsson, Henrik; Drewes Nielsen, Lise

    Globalisation is heavily dependent on physical transport, as people and goods travel over longer distances and with higher frequency. Movement and mobility have become integrated parts of late modern identity and practice, and a state of flux can be sensed everywhere. Bringing together the latest...... for both research and policy. In the final section of the book new visions for research into sustainability and mobility are laid out....

  18. Mobile Apps and Romanian Mobile Devices Users’ Preferences

    OpenAIRE

    Tutunea, Mihaela Filofteia

    2014-01-01

    During the last years the development of mobile technologies generated the formation of a new segment of software industry dedicated to applications for mobile devices. At their turn, mobile apps created new tendencies in software development but also generated important revenues at the level of the specialized industry as well as at global level. From this perspective, the present article, on one hand, describes an image of the mobile infrastructure, mobile applications ...

  19. Mastering jQuery mobile

    CERN Document Server

    Lambert, Chip

    2015-01-01

    You've started down the path of jQuery Mobile, now begin mastering some of jQuery Mobile's higher level topics. Go beyond jQuery Mobile's documentation and master one of the hottest mobile technologies out there. Previous JavaScript and PHP experience can help you get the most out of this book.

  20. Mobility. Snapshot Report, Fall 2011

    Science.gov (United States)

    National Student Clearinghouse, 2011

    2011-01-01

    This paper presents information on student mobility for 2011. It offers data on the following: (1) Mobility Rates by Student Enrollment Status; (2) Mobile Student Enrollment at 2-/4-Year Institutions; and (3) Mobile Student Enrollment at Public/Private Institutions.

  1. Mobility decline in old age

    DEFF Research Database (Denmark)

    Rantakokko, Merja; Mänty, Minna Regina; Rantanen, Taina

    2013-01-01

    Mobility is important for community independence. With increasing age, underlying pathologies, genetic vulnerabilities, physiological and sensory impairments, and environmental barriers increase the risk for mobility decline. Understanding how mobility declines is paramount to finding ways...... to promote mobility in old age....

  2. Libraries and the Mobile Web

    Science.gov (United States)

    Hanson, Cody

    2011-01-01

    In 2011, cell phones and mobile devices are ubiquitous. The vast majority of Americans now own cell phones, and over half of them have mobile access to the Internet through a phone or other mobile device. For libraries to stay relevant, they must be able to offer content and services through the mobile web. In this issue of "Library Technology…

  3. Mobile medical image retrieval

    Science.gov (United States)

    Duc, Samuel; Depeursinge, Adrien; Eggel, Ivan; Müller, Henning

    2011-03-01

    Images are an integral part of medical practice for diagnosis, treatment planning and teaching. Image retrieval has gained in importance mainly as a research domain over the past 20 years. Both textual and visual retrieval of images are essential. In the process of mobile devices becoming reliable and having a functionality equaling that of formerly desktop clients, mobile computing has gained ground and many applications have been explored. This creates a new field of mobile information search & access and in this context images can play an important role as they often allow understanding complex scenarios much quicker and easier than free text. Mobile information retrieval in general has skyrocketed over the past year with many new applications and tools being developed and all sorts of interfaces being adapted to mobile clients. This article describes constraints of an information retrieval system including visual and textual information retrieval from the medical literature of BioMedCentral and of the RSNA journals Radiology and Radiographics. Solutions for mobile data access with an example on an iPhone in a web-based environment are presented as iPhones are frequently used and the operating system is bound to become the most frequent smartphone operating system in 2011. A web-based scenario was chosen to allow for a use by other smart phone platforms such as Android as well. Constraints of small screens and navigation with touch screens are taken into account in the development of the application. A hybrid choice had to be taken to allow for taking pictures with the cell phone camera and upload them for visual similarity search as most producers of smart phones block this functionality to web applications. Mobile information access and in particular access to images can be surprisingly efficient and effective on smaller screens. Images can be read on screen much faster and relevance of documents can be identified quickly through the use of images contained in

  4. Mobility Network and Safety

    Directory of Open Access Journals (Sweden)

    Adriana Galderisi

    2010-04-01

    Full Text Available Mobility network is crucial for ensuring territorial safety with respect to natural and technological hazards. They represent a basic support to community’s everyday life although being exposed elements often characterized by high vulnerability to different hazards and, in the meanwhile, strategic equipments for emergency management. Physical damages or the lack in functioning of those networks may greatly increase the loss of human lives caused by hazardous events as well as produce relevant economic damages at medium and long term. Although the relevance of the mobility networks in assuring territorial safety is at present largely recognized, risk analyses have been long focused on buildings’ vulnerability or, even where they have paid attention to mobility network, they have been mainly focused on the physical damages that a given hazard could may induce on individual elements of such network. It is recent the awareness that mobility network represents a system, characterized by relevant interdependences both among its elements and among network infrastructures and urban systems. Based on these assumptions, this paper points out the heterogeneous aspects of the mobility network vulnerability and their relevance in increasing the overall territorial or urban vulnerability to hazardous events. Therefore, an in-depth investigation of the concept of mobility network vulnerability is provided, in order to highlight the aspects mostly investigated and more recent research perspectives. Finally, a case study in the Campania Region is presented in order to point out how traditional risk analyses, generally referred to individual hazards, can sometimes led to invest in the mobility network improvement or development which, targeted to increase the security of a territory result, on the opposite, in an increase of the territorial vulnerability.

  5. Mobile applications in dermatology.

    Science.gov (United States)

    Brewer, Ann Chang; Endly, Dawnielle C; Henley, Jill; Amir, Mahsa; Sampson, Blake P; Moreau, Jacqueline F; Dellavalle, Robert P

    2013-11-01

    With advancements in mobile technology, cellular phone-based mobile applications (apps) may be used in the practice and delivery of dermatologic care. To identify and categorize the variety of current mobile apps available in dermatology for patients and providers. Dermatology-related search terms were queried in the online app stores of the most commonly used mobile platforms developed by Apple, Android, Blackberry, Nokia, and Windows. Applications were assigned to categories based on description. Popularity, price, and reviews were recorded and target audiences were determined through websites offering online mobile apps. Number, type, and price of mobile apps in dermatology. A total of 229 dermatology-related apps were identified in the following categories: general dermatology reference (61 [26.6%]), self-surveillance/diagnosis (41 [17.9%]), disease guide (39 [17.0%]), educational aid (20 [8.7%]), sunscreen/UV recommendation (19 [8.3%]), calculator (12 [5.2%]), teledermatology (8 [3.5%]), conference (6 [2.6%]), journal (6 [2.6%]), photograph storage/sharing (5 [2.2%]), dermoscopy (2 [0.9%]), pathology (2 [0.9%]), and other (8 [3.5%]). The most reviewed apps included Ultraviolet ~ UV Index (355 reviews), VisualDx (306), SPF (128), iSore (61), and SpotMole (50). There were 209 unique apps, with 17 apps existing on more than 1 operating system. More than half of the apps were offered free of charge (117 [51.1%]). Paid apps (112 [48.9%]) ranged from $0.99 to $139.99 (median, $2.99). Target audiences included patient (117 [51.1%]), health care provider (94 [41.0%]), and both (18 [7.9%]). The widespread variety and popularity of mobile apps demonstrate a great potential to expand the practice and delivery of dermatologic care.

  6. MobilED: a mobile tools and services platform for formal and informal learning

    CSIR Research Space (South Africa)

    Ford, M

    2009-03-01

    Full Text Available MobilED is a South African initiative aimed at designing teaching and learning environments that are meaningfully enhanced with mobile technologies and services. The deliverables are the development of a set of scenarios and guidelines on how mobile...

  7. Mobile Apps and Romanian Mobile Devices Users’ Preferences

    Directory of Open Access Journals (Sweden)

    Mihaela Filofteia TUTUNEA

    2014-10-01

    Full Text Available During the last years the development of mobile technologies generated the formation of a new segment of software industry dedicated to applications for mobile devices. At their turn, mobile apps created new tendencies in software development but also generated important revenues at the level of the specialized industry as well as at global level. From this perspective, the present article, on one hand, describes an image of the mobile infrastructure, mobile applications and their users, on the other hand, includes a study regarding Romanian users of mobile devices in order to identify the types of applications usually employed in mobile activities, the time dedicated to these activities, the number of downloaded applications and their cost levels. We consider that the results of this study may be useful to mobile app users and developers, and mobile device providers.

  8. Mobile phones, social ties, and collective action mobilization in China

    DEFF Research Database (Denmark)

    Liu, Jun

    2017-01-01

    participants for protests, this study observes that mobile communication in China embodies guanxi, the indigenous social tie in Chinese society that introduces reciprocity as an influential facilitator of collective actions. The embedment of reciprocity facilitates the proliferation of mobilizing calls......To provide a better understanding of mobile phones as a recruitment tool in collective actions, this study explores the use of mobile phones for mobilizing protest in China. Using in-depth interviews and investigating four cases in which Chinese people employed mobile devices to recruit......, legitimizes mobilizing appeals, generates obligations and consolidates solidarity for collective actions. The study concludes with a consideration of the relevance of mobile phones for the embedment of reciprocity in social ties in the mobilization of collective action in authoritarian regimes such as China....

  9. Efficient Mobility Management Signalling in Network Mobility Supported PMIPV6

    Directory of Open Access Journals (Sweden)

    Ananthi Jebaseeli Samuelraj

    2015-01-01

    Full Text Available Proxy Mobile IPV6 (PMIPV6 is a network based mobility management protocol which supports node’s mobility without the contribution from the respective mobile node. PMIPV6 is initially designed to support individual node mobility and it should be enhanced to support mobile network movement. NEMO-BSP is an existing protocol to support network mobility (NEMO in PMIPV6 network. Due to the underlying differences in basic protocols, NEMO-BSP cannot be directly applied to PMIPV6 network. Mobility management signaling and data structures used for individual node’s mobility should be modified to support group nodes’ mobility management efficiently. Though a lot of research work is in progress to implement mobile network movement in PMIPV6, it is not yet standardized and each suffers with different shortcomings. This research work proposes modifications in NEMO-BSP and PMIPV6 to achieve NEMO support in PMIPV6. It mainly concentrates on optimizing the number and size of mobility signaling exchanged while mobile network or mobile network node changes its access point.

  10. Mobile systems capability plan

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1996-09-01

    This plan was prepared to initiate contracting for and deployment of these mobile system services. 102,000 cubic meters of retrievable, contact-handled TRU waste are stored at many sites around the country. Also, an estimated 38,000 cubic meters of TRU waste will be generated in the course of waste inventory workoff and continuing DOE operations. All the defense TRU waste is destined for disposal in WIPP near Carlsbad NM. To ship TRU waste there, sites must first certify that the waste meets WIPP waste acceptance criteria. The waste must be characterized, and if not acceptable, subjected to additional processing, including repackaging. Most sites plan to use existing fixed facilities or open new ones between FY1997-2006 to perform these functions; small-quantity sites lack this capability. An alternative to fixed facilities is the use of mobile systems mounted in trailers or skids, and transported to sites. Mobile systems will be used for all characterization and certification at small sites; large sites can also use them. The Carlsbad Area Office plans to pursue a strategy of privatization of mobile system services, since this offers a number of advantages. To indicate the possible magnitude of the costs of deploying mobile systems, preliminary estimates of equipment, maintenance, and operating costs over a 10-year period were prepared and options for purchase, lease, and privatization through fixed-price contracts considered.

  11. Mobile propeller dynamometer validation

    Science.gov (United States)

    Morris, Mason Wade

    With growing interest in UAVs and OSU's interest in propeller performance and manufacturing, evaluating UAV propeller and propulsion system performance has become essential. In attempts to evaluate these propellers a mobile propeller dynamometer has been designed, built, and tested. The mobile dyno has been designed to be cost effective through the ability to load it into the back of a test vehicle to create simulated forward flight characteristics. This allows much larger propellers to be dynamically tested without the use of large and expensive wind tunnels. While evaluating the accuracy of the dyno, several improvements had to be made to get accurate results. The decisions made to design and improve the mobile propeller dyno will be discussed along with attempts to validate the dyno by comparing its results against known sources. Another large part of assuring the accuracy of the mobile dyno is determining if the test vehicle will influence the flow going into the propellers being tested. The flow into the propeller needs to be as smooth and uniform as possible. This is determined by characterizing the boundary layer and accelerated flow over the vehicle. This evaluation was accomplished with extensive vehicle aerodynamic measurements with the use of full-scale tests using a pitot-rake and the actual test vehicle. Additional tests were conducted in Oklahoma State University's low speed wind tunnel with a 1/8-scale model using qualitative flow visualization with smoke. Continuing research on the mobile dyno will be discussed, along with other potential uses for the dyno.

  12. [Mobility and balance].

    Science.gov (United States)

    Schmitt, Katrin; Kressig, Reto W

    2008-08-01

    Quality of life is strongly associated with the mobility of elderly people. Falls often cause restricted mobility, a decline in activities of daily living and an increased risk of institutionalisation. Frailty, commonly associated with aging, is a biologic syndrome of decreased resistance to stressors, resulting from declines across multiple physiological systems. Changes in mobility and gait constitute part of the frailty syndrome. Since more than one third of persons over the age of 65 fall each year, prevention of falls is very important. Already while taking the patients' history special emphasis should be laid on matters associated with an increased risk of falling, such as the use of more than four medications. To assess mobility several brief tests exist (i.e. Timed up & go [17], Walking while Talking [20]) which immediately yield information regarding mobility and falling risk. Patients with poor performance on such tests or those with a history of several falls should undergo a spatio-temporal gait analysis in order to determine a possible cause as well as suitable interventions. Additionally, the objective measurement of temporo-spatial gait parameters under dual task conditions may detect deficits in cognitive function. Several interventions have been shown to have favourable effects on gait stability and the occurrence of falls. Proprioceptive problems can be partially compensated for by wearing special shoes. Also, different movement exercises such as Tai Chi Chuan, Jaques-Dalcroze eurhythmics and social dancing are associated with better balance and gait safety, and a reduction of falls.

  13. Mobility decline in old age

    DEFF Research Database (Denmark)

    Nilsson, C.J.; Siersma, V.; Mänty, Minna Regina

    2014-01-01

    BACKGROUND: Mobility-related fatigue and low socioeconomic position predicts mobility limitations and disability in old age, but the interplay between these two factors is unknown. To evaluate whether mobility-related fatigue is a stronger risk factor for mobility limitations in certain...... estimating equations were performed using four waves of data on 2874 individuals without mobility limitations at baseline from The Danish Intervention Study on Preventive Home Visits. RESULTS: Low socioeconomic position and mobility-related fatigue are risk factors for mobility limitations in old age...... category (high socioeconomic position and no fatigue) of -0.52, psocioeconomic position and -0.96, psocioeconomic position. CONCLUSIONS: Mobility-related fatigue is not a significantly stronger risk factor for subsequent mobility limitations among...

  14. Are mobile phones harmful?

    DEFF Research Database (Denmark)

    Blettner, M; Berg, Gabriele

    2000-01-01

    in cells. Implications of these experimental results on public health concerns are yet unclear. Few epidemiological studies are available on the use of mobile phones or on the radiofrequency exposure and the development of cancer. Most of these studies have no or little quantitative exposure data......There is increasing public interest in health risks of mobile phone use. Although there is a vast body of material on the biological effects of radiofrequency fields, current risk assessment is still limited. The article describes several hypotheses and results of biological effects such as thermal...... effect, genetic and carcinogenic effects and cancer related investigations. Mobile phones transmit and receive waves of frequencies mainly at 800-1800 MHz. Findings on the thermal effect of acute exposure to radiofrequency fields were consistent, resulting in an increase of cellular, tissue or body...

  15. Intensities of Mobility

    DEFF Research Database (Denmark)

    Bissell, David; Vannini, Phillip; Jensen, Ole B.

    2017-01-01

    to complex social and geographical dynamics of transport, housing, lifestyle, and employment. Yet, the experiential dimensions of long-distance commuting have not received the attention that they deserve within research on mobilities. Drawing from fieldwork conducted in Australia, Canada, and Denmark...... this paper aims to further develop our collective understanding of the experiential particulars of long-distance workers or ‘supercommuters’. Rather than focusing on the extensive dimensions of mobilities that are implicated in broad social patterns and trends, our paper turns to the intensive dimensions......This paper explores the intensities of long-distance commuting journeys in order to understand how bodily sensibilities become attuned to the regular mobilities which they undertake. More people are travelling farther to and from work than ever before, owing to a variety of factors which relate...

  16. Supplier Resource Mobilization

    DEFF Research Database (Denmark)

    Ellegaard, Chris; Kragh, Hanne; Andersen, Poul Houman

    Companies that wish to draw benefit from supplier resources beyond standard market offerings must actively make an effort to mobilize these resources. This task has been a major topic of interest in the management literature, but the continued reported problems of companies attempting to influence...... suppliers suggests that the knowledge of this challenge is yet inadequate. The purpose of this paper is therefore to establish a research agenda on supplier resource mobilization that can improve this knowledge. To fulfill the purpose we review the management literature and identify the most prevalent...... theoretical perspectives. This review, synthesis, and resultant discussion allow us to propose that future research should look closer at the resource activation process on the supplier side, the role of the buyer-supplier relationship in resource mobilization, and the approach of the buying company...

  17. Graphene mobility mapping

    DEFF Research Database (Denmark)

    Buron, Jonas Christian Due; Pizzocchero, Filippo; Jepsen, Peter Uhd

    2015-01-01

    Carrier mobility and chemical doping level are essential figures of merit for graphene, and large-scale characterization of these properties and their uniformity is a prerequisite for commercialization of graphene for electronics and electrodes. However, existing mapping techniques cannot directly...... assess these vital parameters in a non-destructive way. By deconvoluting carrier mobility and density from non-contact terahertz spectroscopic measurements of conductance in graphene samples with terahertz-transparent backgates, we are able to present maps of the spatial variation of both quantities over...... graphene indicates dominance by charged scatterers. Unexpectedly, significant variations in mobility rather than doping are the cause of large conductance inhomogeneities, highlighting the importance of statistical approaches when assessing large-area graphene transport properties....

  18. Mobile Probing and Probes

    DEFF Research Database (Denmark)

    Duvaa, Uffe; Ørngreen, Rikke; Weinkouff Mathiasen, Anne-Gitte

    2013-01-01

    Mobile probing is a method, developed for learning about digital work situations, as an approach to discover new grounds. The method can be used when there is a need to know more about users and their work with certain tasks, but where users at the same time are distributed (in time and space......). Mobile probing was inspired by the cultural probe method, and was influenced by qualitative interview and inquiry approaches. The method has been used in two subsequent projects, involving school children (young adults at 15-17 years old) and employees (adults) in a consultancy company. Findings point...... to mobile probing being a flexible method for uncovering the unknowns, as a way of getting rich data to the analysis and design phases. On the other hand it is difficult to engage users to give in depth explanations, which seem easier in synchronous dialogs (whether online or face2face). The development...

  19. Mobile Probing and Probes

    DEFF Research Database (Denmark)

    Duvaa, Uffe; Ørngreen, Rikke; Weinkouff, Anne-Gitte

    2012-01-01

    and space). Mobile probing was inspired by the cultural probe method, and was influenced by qualitative interview and inquiry approaches. The method has been used in two subsequent projects, involving school children (young adults at 15-17 years old) and employees (adults) in a consultancy company. Findings......Mobile probing is a method, which has been developed for learning about digital work situations, as an approach to discover new grounds. The method can be used when there is a need to know more about users and their work with certain tasks, but where users at the same time are distributed (in time...... point to mobile probing being a flexible method for uncovering the unknowns, as a way of getting rich data to the analysis and design phases. On the other hand it is difficult to engage users to give in depth explanations, which seem easier in synchronous dialogs (whether online or face2face...

  20. Identification and Characterization of the Lamprey High-Mobility Group Box 1 Gene

    Science.gov (United States)

    Pang, Yue; Xiao, Rong; Liu, Xin; Li, Qingwei

    2012-01-01

    High-mobility group box 1 (HMGB1), a highly conserved DNA-binding protein, plays an important role in maintaining nucleosome structures, transcription, and inflammation. We identified a homolog of HMGB1 in the Japanese lamprey (Lampetra japonica). The Lampetra japonica HMGB1 gene (Lj-HMGB1) has over 70% sequence identity with its homologs in jawed vertebrates. Despite the reasonably high sequence identity with other HMGB1 proteins, Lj-HMGB1 did not group together with these proteins in a phylogenetic analysis. We examined Lj-HMGB1 expression in lymphocyte-like cells, and the kidneys, heart, gills, and intestines of lampreys before and after the animals were challenged with lipopolysaccharide (LPS) and concanavalin A (ConA). Lj-HMGB1 was initially expressed at a higher level in the heart, but after treatment with LPS and ConA only the gills demonstrated a significant up-regulation of expression. The recombinant Lj-HMGB1 (rLj-HMGB1) protein bound double-stranded DNA and induced the proliferation of human adenocarcinoma cells to a similar extent as human HMGB1. We further revealed that Lj-HMGB1 was able to induce the production of tumor necrosis factor-α (TNF-α), a pro-inflammatory mediator, in activated human acute monocytic leukemia cells. These results suggest that lampreys use HMGB1 to activate their innate immunity for the purpose of pathogen defense. PMID:22563397

  1. Identification and characterization of the lamprey high-mobility group box 1 gene.

    Directory of Open Access Journals (Sweden)

    Yue Pang

    Full Text Available High-mobility group box 1 (HMGB1, a highly conserved DNA-binding protein, plays an important role in maintaining nucleosome structures, transcription, and inflammation. We identified a homolog of HMGB1 in the Japanese lamprey (Lampetra japonica. The Lampetra japonica HMGB1 gene (Lj-HMGB1 has over 70% sequence identity with its homologs in jawed vertebrates. Despite the reasonably high sequence identity with other HMGB1 proteins, Lj-HMGB1 did not group together with these proteins in a phylogenetic analysis. We examined Lj-HMGB1 expression in lymphocyte-like cells, and the kidneys, heart, gills, and intestines of lampreys before and after the animals were challenged with lipopolysaccharide (LPS and concanavalin A (ConA. Lj-HMGB1 was initially expressed at a higher level in the heart, but after treatment with LPS and ConA only the gills demonstrated a significant up-regulation of expression. The recombinant Lj-HMGB1 (rLj-HMGB1 protein bound double-stranded DNA and induced the proliferation of human adenocarcinoma cells to a similar extent as human HMGB1. We further revealed that Lj-HMGB1 was able to induce the production of tumor necrosis factor-α (TNF-α, a pro-inflammatory mediator, in activated human acute monocytic leukemia cells. These results suggest that lampreys use HMGB1 to activate their innate immunity for the purpose of pathogen defense.

  2. Epochal neuroinflammatory role of high mobility group box 1 in central nervous system diseases

    Directory of Open Access Journals (Sweden)

    Seidu A. Richard

    2017-05-01

    Full Text Available The central nervous system (CNS is enriched with a developed reaction reserve dubbed “neuroinflammation”, which facilitates it to cope with pathogens, toxins, traumata and degeneration. Inflammation is a significant biological activity in reaction to injury, infection, and trauma agonized by cells or tissues. A positive inflammatory reaction mechanism removes attacking pathogens, initiating wound healing and angiogenesis. The High Mobility Group Box 1 (HMGB1 protein is abundant and ubiquitous nuclear proteins that bind to DNA, nucleosome and other multi-protein complexes in a dynamic and reversible fashion to regulate DNA processing in the context of chromatin. Complex genetic and physiological variations as well as environmental factors that drive emergence of chromosomal instability, development of unscheduled cell death, skewed differentiation, and altered metabolism are central to the pathogenesis of human diseases and disorders. HMGB1 protein, senses and coordinates the cellular stress response and plays a critical role not only inside of the cell as a DNA chaperone, chromosome guardian, autophagy sustainer, and protector from apoptotic cell death, but also outside the cell as the prototypic damage associated molecular pattern molecule (DAMP. This DAMP, in conjunction with other factors such as cytokine, chemokine, and growth factor activity, orchestrating the inflammatory and immune response. All of these characteristics make HMGB1 a critical molecular target in multiple human diseases including infectious diseases, ischemia, immune disorders, neurodegenerative diseases, metabolic disorders, and cancer. With regards to these various disease condition above, our review focus on the role of HMGB1 and CNS Diseases.

  3. Effects of urban land-use on largescale stonerollers in the Mobile River Basin, Birmingham, AL.

    Science.gov (United States)

    Iwanowicz, D; Black, M C; Blazer, V S; Zappia, H; Bryant, W

    2016-04-01

    During the spring and fall of 2001 and the spring of 2002 a study was conducted to evaluate the health of the largescale stoneroller (Campostoma oligolepis) populations in streams along an urban land-use gradient. Sites were selected from a pool of naturally similar sub-basins (eco-region, basin size, and geology) of the Mobile River basin (MRB), using an index of urban intensity derived from infrastructure, socioeconomic, and land-use data. This urban land-use gradient (ULUG) is a multimetric indicator of urban intensity, ranging from 0 (background) to 100 (intense urbanization). Campostoma sp. have been used previously as indicators of stream health and are common species found in all sites within the MRB. Endpoints used to determine the effects of urban land-use on the largescale stoneroller included total glutathione, histology, hepatic apoptosis, condition factor and external lesions. Liver glutathione levels were positively associated with increasing urban land-use (r(2) = 0.94). Histopathological examination determined that some abnormalities and lesions were correlated with the ULUG and generally increased in prevalence or severity with increasing urbanization. Liver macrophage aggregates were positively correlated to the ULUG. The occurrence of nucleosomal ladders (indicating apoptotic cell death) did not correspond with urban intensity in a linear fashion. Apoptosis, as well as prevalence and severity of a myxozoan parasite, appeared to have a hormetic dose-response relationship. The majority of the biomarkers suggested fish health was compromised in areas where the ULUG ≥ 36.

  4. The Space Mobile Network

    Science.gov (United States)

    Israel, David

    2017-01-01

    The definition and development of the next generation space communications and navigation architecture is underway. The primary goals are to remove communications and navigations constraints from missions and to enable increased autonomy. The Space Mobile Network (SMN) is an architectural concept that includes new technology and operations that will provide flight systems with an similar user experience to terrestrial wireless mobile networks. This talk will describe the SMN and its proposed new features, such as Disruption Tolerant Networking (DTN), optical communications, and User Initiated Services (UIS).

  5. To practice mobility

    DEFF Research Database (Denmark)

    Winther, Ida Wentzel

    2015-01-01

    Children’s perspectives are practically absent in new mobility studies. In this article, I wish to describe and analyze how a number of children handle having to move between homes, parents and siblings, and how they practically, emotionally and socially navigate in this changeable landscape. My...... aim is to explore mobility as an embodied and emotional practice in which children employ different strategies. I focus on bodily micro practices, routines and coping strategies, the intermediate space that occurs on their continual journeys, and the feeling of being dispensable. It is an ethnographic...

  6. Autonomous mobile robot teams

    Science.gov (United States)

    Agah, Arvin; Bekey, George A.

    1994-01-01

    This paper describes autonomous mobile robot teams performing tasks in unstructured environments. The behavior and the intelligence of the group is distributed, and the system does not include a central command base or leader. The novel concept of the Tropism-Based Cognitive Architecture is introduced, which is used by the robots in order to produce behavior transforming their sensory information to proper action. The results of a number of simulation experiments are presented. These experiments include worlds where the robot teams must locate, decompose, and gather objects, and defend themselves against hostile predators, while navigating around stationary and mobile obstacles.

  7. Professional mobile application development

    CERN Document Server

    McWherter, Jeff

    2012-01-01

    Create applications for all major smartphone platforms Creating applications for the myriad versions and varieties of mobile phone platforms on the market can be daunting to even the most seasoned developer. This authoritative guide is written in such as way that it takes your existing skills and experience and uses that background as a solid foundation for developing applications that cross over between platforms, thereby freeing you from having to learn a new platform from scratch each time. Concise explanations walk you through the tools and patterns for developing for all the mobile platfo

  8. Urban Mobility in Indonesia

    OpenAIRE

    2011-01-01

    The April 2011 edition of Prakarsa has the theme “Urban Mobility”. In its pages you will find both an overview of the latest international approaches for achieving mobility and an examination of mobility issues in select Indonesian cities. Feature articles include “An Introduction to Urban Mobility” by Peter Midgley; “Freeing Jakarta From Gridlock” by Danang Parikesit; “The Surabaya Story: Problems and Solutions for Improving Urban Mobility” by William Vincent; and “Solving Bogor's ‘Angkot Pr...

  9. Mobile light in roses

    OpenAIRE

    Marissen, A.; Snel, J.F.H.; Elings, A.; Warmenhoven, M.G.

    2006-01-01

    In order to quantify the claimed differences between mobile and static lighting, and to clarify the mechanisms causing these differences, an experiment with rose `First Red¿ was carried out. In a greenhouse compartment half of the area was equipped with mobile lamps, and the other half with static lamps. By mounting the lamps in the centre of the greenhouse, a light gradient was achieved in both treatments. Experimental plots were chosen with daily light integrals of 1 to 3 and 3 to 5 mol m-2...

  10. Correlation ion mobility spectroscopy

    Science.gov (United States)

    Pfeifer, Kent B [Los Lunas, NM; Rohde, Steven B [Corrales, NM

    2008-08-26

    Correlation ion mobility spectrometry (CIMS) uses gating modulation and correlation signal processing to improve IMS instrument performance. Closely spaced ion peaks can be resolved by adding discriminating codes to the gate and matched filtering for the received ion current signal, thereby improving sensitivity and resolution of an ion mobility spectrometer. CIMS can be used to improve the signal-to-noise ratio even for transient chemical samples. CIMS is especially advantageous for small geometry IMS drift tubes that can otherwise have poor resolution due to their small size.

  11. Mobile network maintenance (GSM)

    CERN Multimedia

    IT Department

    2009-01-01

    Maintenance work will be carried out on the CERN mobile network infrastructure (GSM) on the 23 and 24 July from 6 p.m. to 6 a.m. in order to replace discontinued equipment and to increase the bandwidth capacity of the GSM mobile network. All CERN GSM emitters (40 units) will be moved one by one to the new infrastructure during the maintenance. The call of a user connected to an emitter at the time of its maintenance will be cut off. However, the general overlapping of the GSM radio coverage should mean that users are able immediately to call again should their call be interrupted. IT/CS/CS

  12. Lazy Mobile Intruders

    DEFF Research Database (Denmark)

    Mödersheim, Sebastian Alexander; Nielson, Flemming; Nielson, Hanne Riis

    We present a new technique for analyzing platforms that execute potentially malicious code, such as web-browsers, mobile phones, or virtualized infrastructures. Rather than analyzing given code, we ask what code an intruder could create to break a security goal of the platform. To avoid searching...... a small, abstract formalism that models the essence of mobile code. We provide a decision procedure for security against arbitrary intruder ambients when the honest ambients can only perform a bounded number of steps and without path constraints in communication....

  13. Segway robotic mobility platform

    Science.gov (United States)

    Nguyen, Hoa G.; Morrell, John; Mullens, Katherine D.; Burmeister, Aaron B.; Miles, Susan; Farrington, Nathan; Thomas, Kari M.; Gage, Douglas W.

    2004-12-01

    The Segway Robotic Mobility Platform (RMP) is a new mobile robotic platform based on the self-balancing Segway Human Transporter (HT). The Segway RMP is faster, cheaper, and more agile than existing comparable platforms. It is also rugged, has a small footprint, a zero turning radius, and yet can carry a greater payload. The new geometry of the platform presents researchers with an opportunity to examine novel topics, including people-height sensing and actuation modalities. This paper describes the history and development of the platform, its characteristics, and a summary of current research projects involving the platform at various institutions across the United States.

  14. Profiling the Mobile Customer

    DEFF Research Database (Denmark)

    Jessen, Pernille Wegener; King, Nancy J.

    2010-01-01

    Mobile customers are increasingly being tracked and profiled by behavioural advertisers to enhance delivery of personalized advertising. This type of profiling relies on automated processes that mine databases containing personally-identifying or anonymous consumer data, and it raises a host...... of significant concerns about privacy and data protection. This second article in a two part series on "Profiling the Mobile Customer" explores how to best protect consumers' privacy and personal data through available mechanisms that include industry self-regulation, privacy-enhancing technologies...

  15. An electromagnetic perpetuum mobile?

    OpenAIRE

    Grøn, Øyvind; Næss, Sigurd Kirkevold

    2008-01-01

    A charge moving freely in orbit around the Earth radiates according to Larmor's formula. If the path is closed, it would constitute a perpetuum mobile. The solution to this energy paradox is found in an article by C. M. DeWitt and B. DeWitt from 1964. The main point is that the equation of motion of a radiating charge is modified in curved spacetime. In the present article we explain the physics behind this modification, and use the generalized equation to solve the perpetuum mobile paradox.

  16. Protocols in Mobile Electronic Commerce

    OpenAIRE

    Mr. Vivek B. Patil; Mr.Deepak G. Awate

    2017-01-01

    Mobile-commerce, also known by other terms such as M-Commerce or m-Commerce, is the capability to follow commerce with the help of a hand held devise like mobile phone, a Personal digital assistant PDA, a smart phone, or other emerging mobile equipment such as dash top mobile devices. Since decade Mobile-commerce is being expressed as follows: Mobile-commerce is where any transaction, involving the transfer of rights or ownership to use goods and services, which is initialized and/or processe...

  17. Instant XenMobile MDM

    CERN Document Server

    Lakhani, Aamir

    2013-01-01

    Get to grips with a new technology, understand what it is and what it can do for you, and then get to work with the most important features and tasks. Written in a user friendly style, this guide will get readers up and running with XenMobile MDM.If you want to run your mobile enterprises on XenMobile, or work on a BYOD strategy within your organization, then this is the ideal book for you. XenMobile MDM comprehensively explores how to set up and use XenMobile to provision, secure, and manage mobile devices.

  18. Mobile Service Platform: A Middleware for Nomadic Mobile Service Provisioning

    NARCIS (Netherlands)

    van Halteren, Aart; Pawar, P.

    Nowadays mobile devices are characterized by higher processing power, lower costs, multiple network interfaces, ability to support multiple auxiliary devices and connect to the Internet using a wireless network. The applications collecting statistics concerning the mobile user, computational and

  19. Mobile agents basic concepts, mobility models, and the tracy toolkit

    CERN Document Server

    Braun, Peter

    2005-01-01

    Mobile agents are software nomads that act as your personal representative, working autonomously through networks. They are able to visit network nodes directly using available computing power and are not limited by platform. This emerging field is now poised to become a cornerstone for new Web-based ubiquitous computing environments. Mobile Agents provides a practical introduction to mobile agent technology and surveys the state of the art in mobile agent research. Students and researchers can use the book as an introduction to the concepts and possibilities of this field and as an overview of ongoing research. Developers can use it to identify the capabilities of the technology to decide if mobile agents are the right solution for them. Practioners can also gain hands-on experience in programming mobile agents through exploration of the source code for a complete mobile agent environment available through the companion website.*Summarizes the state of the art in mobile agent research*Identifies the benefits...

  20. On the extent of researcher mobility and indicators for mobility

    Energy Technology Data Exchange (ETDEWEB)

    Barriere, S.; Baard, P.; Karlsson, S.; Jacobsson, C.; Tumpane, J.

    2016-07-01

    Researchers' mobility is significant for the quality of research, although mobility does not necessarily have an intrinsic value. Sweden appears to have a lover degree of mobility when compared to a selection of successful research countries. The Swedish Research Council shall, according to the instruction from the government , promote researcher mobility. This study explores job-mobility within the Swedish academic system based on register studies and draw some conclusions on the degree of mobility in different different disciplines, differences between men and women as well as on the development of comprehensive and easy to follow indicators for mobility. The study intends to provide support for recommendations on researcher mobility within the Research Council's overarching goal to promote excellence in research. (Author)