Analysis of chromatin structure by in vivo formaldehyde cross-linking
Analysis of chromatin structure by in vivo formaldehyde cross-linking
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DOI：
10.1006/meth.1996.0407
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发表时间：
1997-02-01
期刊：
影响因子：
--
通讯作者：
Paro, R
中科院分区：
文献类型：
--
作者：
Orlando, V;Strutt, H;Paro, R
最近的研究表明，高阶染色质结构在许多发育上重要的基因调控机制中发挥着重要的作用。特别是，在酵母和果蝇等遗传模型系统中的分析发现了参与染色质结构调节的新蛋白质。这些蛋白质中的许多并不直接与DNA结合，而是在大的多聚体复合体中相互作用。为了确定受这些多蛋白复合体调控的DNA元件，必须设计出替代DNA-蛋白质分析标准方法的方法。在这里，我们提出了一种方法，通过在体内用甲醛交联细胞来保存高阶染色质结构的结构。然后使用免疫沉淀策略来识别感兴趣的染色体蛋白质的DNA靶标。这种方法可以用来研究蛋白质在延伸的染色体区域上的高分辨率分布。(C)1997年学术出版社。
Recent advances leave no doubt that higher order chromatin structures play a fundamental role in many developmentally important mechanisms of gene regulation. In particular analyses in genetic model systems like yeast and Drosophila uncovered novel proteins that are involved in the regulation of chromatin structures. Many of these proteins do not bind directly to DNA but interact in large multimeric complexes. To identify the DNA elements regulated by these multiprotein complexes, alternative approaches to the standard methods of DNA-protein analysis had to be devised. Here we present a method that preserves the architecture of the higher order chromatin structures by crosslinking cells in vivo with formaldehyde. An immunoprecipitation strategy is then used to identify the DNA targets of chromosomal proteins of interest. This method can be applied to study the distribution of proteins at high resolution over extended chromosomal regions. (C) 1997 Academic Press.