ChAP-MS: a method for identification of proteins and histone posttranslational modifications at a single genomic locus.
ChAP-MS: a method for identification of proteins and histone posttranslational modifications at a single genomic locus.
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DOI：
10.1016/j.celrep.2012.06.019
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发表时间：
2012-07-26
期刊：
影响因子：
8.8
通讯作者：
Tackett AJ
中科院分区：
文献类型：
--
作者：
Byrum SD;Raman A;Taverna SD;Tackett AJ
表观基因组学的领域已经被染色质免疫沉淀方法所改变，该方法提供了将定义的蛋白质或后修饰的蛋白质定位到特定的染色体位点。虽然这些方法帮助我们概念化表观遗传机制，但该领域受到无法以无偏见的方式定义特定基因组位点的蛋白质组和组蛋白修饰等特征的限制。我们开发了一种无偏的方法，即一个独特的天然基因组位点被分离，然后通过高分辨率的蛋白质组学鉴定特异性相关的蛋白质和组蛋白翻译后修饰。该染色质亲和纯化质谱（ChAP-MS）技术用于在转录活性和抑制条件下特异性富集GAL 1染色质的约1，000个碱基对部分，并鉴定特异性结合的蛋白质和组蛋白翻译后修饰。ChAP-MS应该对转录的调控机制产生前所未有的洞察力，并帮助确定表观遗传控制染色质功能的因素。
The field of epigenomics has been transformed by chromatin immunoprecipitation approaches that provide for the localization of a defined protein or posttranslationally-modified protein to specific chromosomal sites. While these approaches have helped us conceptualize epigenetic mechanisms, the field has been limited by the inability to define features like the proteome and histone modifications at a specific genomic locus in an unbiased manner. We developed an unbiased approach whereby a unique native genomic locus was isolated, which was followed by high resolution proteomic identification of specifically associated proteins and histone posttranslational modifications. This Chromatin Affinity Purification with Mass Spectrometry (ChAP-MS) technique was used to specifically enrich a ~1,000 base-pair section of GAL1 chromatin under transcriptionally active and repressive conditions, and to identify the specifically bound proteins and histone posttranslational modifications. ChAP-MS should yield unprecedented insight into the regulatory mechanisms of transcription and help identify factors that epigenetically control chromatin function.
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DOI：
10.1007/978-1-61779-477-3_10
发表时间：
2012
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Byrum, Stephanie;Smart, Sherri K;Larson, Signe;Tackett, Alan J
通讯作者：
Tackett, Alan J
影响因子：
5.3
作者：
Griesenbeck, J;Boeger, H;Kornberg, RD
通讯作者：
Kornberg, RD
影响因子：
3.7
作者：
Bock, Ina;Dhayalan, Arunkumar;Jeltsch, Albert
通讯作者：
Jeltsch, Albert
影响因子：
10.5
作者：
Akiyoshi, Bungo;Nelson, Christian R.;Biggins, Sue
通讯作者：
Biggins, Sue
影响因子：
4.4
作者：
Tackett, AJ;DeGrasse, JA;Chait, BT
通讯作者：
Chait, BT