Abstract:
DNA methylation and posttranslational modifications of histones instruct gene expression in eukaryotes. Besides canonical histones, histone variants also play a critical role in transcriptional regulation. One of the best studied histone variants in plants is H2A.Z whose removal from gene bodies correlates with increased transcriptional activity. The eviction of H2A.Z is regulated by environmental cues such as increased ambient temperatures, and current models suggest that H2A.Z functions as a transcriptional buffer preventing environmentally responsive genes from undesired activation. To monitor temperature-dependent H2A.Z dynamics, chromatin immunoprecipitation (ChIP) of H2A.Z-occupied DNA can be performed. The following protocol describes a quick and easy ChIP approach to study in vivo H2A.Z occupancy.
摘要:
组蛋白的DNA甲基化和翻译后修饰指示真核生物中的基因表达。除了规范的组蛋白,组蛋白变体在转录调控中也起着关键作用。植物中研究最好的组蛋白变体之一是H2A。从基因体中去除的Z与增加的转录活性相关。H2A的驱逐。Z受环境线索的调节,如环境温度升高,目前的模型表明H2A。Z起转录缓冲液的作用,防止环境应答基因不希望的激活。监测与温度相关的H2A。Z动力学,H2A的染色质免疫沉淀(ChIP)。可以进行Z-占据的DNA。以下方案描述了一种快速简便的ChIP方法来研究体内H2A。Z占用率。