Protein Landscape at Drosophila melanogaster Telomere-Associated Sequence Repeats
Protein Landscape at Drosophila melanogaster Telomere-Associated Sequence Repeats
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DOI：
10.1128/mcb.00010-12
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发表时间：
2012-06-01
影响因子：
5.3
通讯作者：
Kingston, Robert E.
中科院分区：
文献类型：
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作者：
Antao, Jose M.;Mason, James M.;Kingston, Robert E.
在每个基因组位点上结合的特定蛋白质组对调节过程和细胞的身份起决定性作用。了解特定基因座的功能需要了解哪些因素与该基因座相互作用，以及蛋白质组成在不同细胞类型中或在响应内部和外部信号时如何变化。分离染色质片段的蛋白质组学分析（PICh）被开发为一种靶向、纯化和鉴定与特定位点相关的蛋白质的工具，并被证明可以纯化人类端粒染色质。在这里，我们开发了这种方法来鉴定与果蝇端粒相关序列（TAS）重复序列相互作用的蛋白质。通过染色质免疫沉淀验证了几个纯化因子是新的tas结合蛋白，并且通过使用基因测试证实了Brahma复合体是端粒位置效应的显性修饰因子。这些结果提供了将PICh方案应用于序列比人类端粒更复杂的位点的有效性的信息，并鉴定了与TAS重复序列结合的蛋白质，这可能有助于TAS生物学和染色质沉默。
The specific set of proteins bound at each genomic locus contributes decisively to regulatory processes and to the identity of a cell. Understanding of the function of a particular locus requires the knowledge of what factors interact with that locus and how the protein composition changes in different cell types or during the response to internal and external signals. Proteomic analysis of isolated chromatin segments (PICh) was developed as a tool to target, purify, and identify proteins associated with a defined locus and was shown to allow the purification of human telomeric chromatin. Here we have developed this method to identify proteins that interact with the Drosophila telomere-associated sequence (TAS) repeats. Several of the purified factors were validated as novel TAS-bound proteins by chromatin immunoprecipitation, and the Brahma complex was confirmed as a dominant modifier of telomeric position effect through the use of a genetic test. These results offer information on the efficacy of applying the PICh protocol to loci with sequence more complex than that found at human telomeres and identify proteins that bind to the TAS repeats, which might contribute to TAS biology and chromatin silencing.