Identifying the Epigenetic Regulation of Arsenic Exposure

 描述（由申请人提供）：大量大分子蛋白质相互作用必须在染色质上正确发生，以驱动染色体生物学的功能方面，如基因转录、DNA复制、重组、修复和姐妹染色单体分离。分析蛋白质如何在体内与染色质相互作用，以指导这些活动仍然是一个重大的挑战，由于其协会的时间和动态的性质。我们最近开发了一种称为染色质亲和纯化与质谱（ChAP-MS）的技术，该技术提供了一个本地1 kb的染色体部分的富集，用于蛋白质相互作用和相关的组蛋白翻译后修饰（PTM）的位点特异性鉴定。在这项拨款申请中，我们计划使用这种尖端的方法来定义组蛋白PTM和蛋白质调节转录在砷反应位点的芽殖酵母。我们假设，ChAP-MS将提供一个全面和公正的鉴定所有组蛋白修饰和蛋白质调节转录的砷基因座在S。啤酒。我们将采用以下目的来验证这一假设：使用ChAP-MS来确定S.啤酒。

 DESCRIPTION (provided by applicant): A multitude of macromolecular protein interactions must properly occur on chromatin to drive functional aspects of chromosome biology like gene transcription, DNA replication, recombination, repair and sister chromatid segregation. Analyzing how proteins interact in vivo with chromatin to direct these activities remains a significant challenge due to the temporal and dynamic nature of their associations. We have recently developed a technique termed Chromatin Affinity Purification with Mass Spectrometry (ChAP-MS) that provides for the enrichment of a native 1 kb section of a chromosome for site-specific identification of protein interactions and associated histone posttranslational modifications (PTMs). In this grant application, we plan to use this cutting-edge approach to define the histone PTMs and proteins regulating transcription at the arsenic response locus in budding yeast. We hypothesize that ChAP-MS will provide for a comprehensive and unbiased identification of all histone modifications and proteins regulating transcription at the arsenic locus in S. cerevisiae. We will pursue the following Aim to test this hypothesis: Use ChAP-MS to define the histone PTMs and proteins regulating transcription at the arsenic response locus in S. cerevisiae.