The chromatin remodeling protein, SRCAP, is critical for deposition of the histone variant H2A.Z at promoters

被引:124
作者
Wong, Madeline M. [1 ]
Cox, Linda K. [1 ]
Chrivia, John C. [1 ]
机构
[1] St Louis Univ, Sch Med, Dept Pharmacol & Physiol Sci, St Louis, MO 63104 USA
关键词
D O I
10.1074/jbc.M703418200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Studies in Saccharomyces cerevisiae indicate the histone variant H2A. Z is deposited at promoters by the chromatin remodeling protein Swr1 and plays a critical role in the regulation of transcription. In higher eukaryotes, however, little is known about the distribution, method of deposition, and function of H2A. Z at promoters. Using biochemical studies, we demonstrated previously that SRCAP ( SNF-2-related CREB-binding protein activator protein), the human ortholog of Swr1, could catalyze deposition of H2A. Z into nucleosomes. To address whether SRCAP directs H2A. Z deposition in vivo, promoters targeted by SRCAP were identified by a chromatin immunoprecipitation ( ChIP)-on-chip assay. ChIP assays on a subset of these promoters confirmed the presence of SRCAP on inactive and active promoters. The highest levels of SRCAP were observed on the active SP-1, G3BP, and FAD synthetase promoters. Detailed analyses of these promoters indicate sites of SRCAP binding overlap or occur adjacent to the sites of H2A. Z deposition. Knockdown of SRCAP levels using siRNA resulted in loss of SRCAP at these promoters, decreased deposition of H2A. Z and acetylated H2A. Z, and a decrease in levels of SP-1, G3BP, and FAD synthetase mRNA. Thus, these studies provide the first evidence that SRCAP is recruited to promoters and is critical for the deposition of H2A.
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页码:26132 / 26139
页数:8
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