Analysis of the Heat Shock Factor Complex in Mammalian HSP70 Promoter.
Analysis of the Heat Shock Factor Complex in Mammalian HSP70 Promoter.
复制标题
哺乳动物 HSP70 启动子中热休克因子复合物的分析。
DOI：
10.1007/978-1-4939-2522-3_4
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Nakai A.
中科院分区：
文献类型：
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作者：
Fujimoto M;Takii R;Hayashida N;Nakai A.
热休克反应的特征是诱导热休克蛋白（HSPs）的产生，是调节细胞蛋白质稳态能力的重要机制之一。在哺乳动物中，热休克因子1（HSF 1）在非应激和应激细胞中转录调节热休克蛋白的表达。最近的报道表明，HSF 1-RPA复合物组成性地获得进入核小体DNA的部分通过招募组蛋白伴侣和染色质重塑组分。在这里，我们描述的策略，以取代内源性HSF 1异位表达的HSF 1或其突变体，并检测HSF 1转录复合物，包括RPA在体内的两个热休克反应元件位于靠近在人类或小鼠HSP 70启动子染色质免疫沉淀法具有高灵敏度和特异性。
The heat shock response is characterized by the induction of heat shock proteins (HSPs) and is one of prominent mechanisms that regulate proteostasis capacity in the cell. In mammals, heat shock factor 1 (HSF1) regulates the expression of HSPs transcriptionally in both unstressed and stressed cells. Recent reports show that the HSF1-RPA complex constitutively gains access to nucleosomal DNA in part by recruiting a histone chaperone and a chromatin-remodeling component. Here, we describe the strategies to substitute endogenous HSF1 with ectopically expressed HSF1 or its mutant and to detect the occupancy of HSF1 transcription complex including RPA in vivo on two heat shock response elements located close together in the human or mouse HSP70 promoters by chromatin immunoprecipitation assay with high sensitivity and specificity.