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有读书笔记有附件Genome-Wide Kinetics of Nucleosome Turnover Determined by Metabolic Labeling of Histones

热4 yonghao616 添加于 2010-6-24 13:07 | 1862 次阅读 | 0 个评论

  •  作 者

    Deal RB, Henikoff JG, Henikoff S

  •  摘 要

    Nucleosome disruption and replacement are crucial activities that maintain epigenomes, but these highly dynamic processes have been difficult to study. Here, we describe a direct method for measuring nucleosome turnover dynamics genome-wide. We found that nucleosome turnover is most rapid over active gene bodies, epigenetic regulatory elements, and replication origins in Drosophila cells. Nucleosomes turn over faster at sites for trithorax-group than polycomb-group protein binding, suggesting that nucleosome turnover differences underlie their opposing activities and challenging models for epigenetic inheritance that rely on stability of histone marks. Our results establish a general strategy for studying nucleosome dynamics and uncover nucleosome turnover differences across the genome that are likely to have functional importance for epigenome maintenance, gene regulation, and control of DNA replication.

  •  详细资料

    • 文献种类:期刊
    • 期刊名称: Science
    • 期刊缩写: Science
    • 期卷页: 2010年  第328卷 第5982期 1161-1164页
    • ISBN: 0036-8075

  • 相关链接 DOI URL 

  •  附 件

    PDF附件Genome-Wide Kinetics of Nucleosome Turnover Determined by Metabolic Labeling of Histones 

  •  yonghao616 的文献笔记  订阅

    核小体的解离-重新组合（即转换，turnover）的实时捕捉新技术

    Deal等研发了一项研究核小体实时转换的新技术，即所谓的CATCH-IT（covalent attachment of tags to capture histones and identify turnover）。他们以黑腹果蝇S2细胞为研究对象，技术流程如下：用甲硫氨酸（Met）的替代氨基酸azidohomoalanine（Aha）处理S2细胞，然后当组蛋白翻译时Aha就会参入进新合成的组蛋白中；用生物素（biotin）连接Aha，这样发生新的转换的核小体就会被生物素标记；将染色质打碎成为单个的核小体，用抗生蛋白链菌素（streptavidin）磁珠提取生物素标记的刚发生转换（新合成）的核小体；将这些核小体上的DNA链和高密度的基因芯片杂交，获得这些DNA的信息。这样就可以得知那些DNA区段上的核小体发生了新的解离-合成过程。

    用这一技术，Deal等发现在果蝇细胞中活性基因、表观调控元件和复制区域的核小体会发生转换。

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