The chromatin remodeling protein, SRCAP, is critical for deposition of the histone variant H2A.Z at promoters.
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.Z.
Wong
Madeline M
MM
Department of Pharmacological and Physiological Science, Saint Louis University School of Medicine, Saint Louis, Missouri 63104, USA.
Cox
Linda K
LK
Chrivia
John C
JC
D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D045744:Cell Line, Tumor; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects / Q000502:physiology*; D047369:Chromatin Immunoprecipitation; D004265:DNA Helicases; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D046228:Microarray Analysis; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism*; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics; D000075223:Poly-ADP-Ribose Binding Proteins; D020365:RNA Helicases; D000067776:RNA Recognition Motif Proteins; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D020218:Response Elements / Q000502:physiology*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000096:biosynthesis / Q000235:genetics
D002352:Carrier Proteins; D006657:Histones; D009707:Nucleosomes; D000075223:Poly-ADP-Ribose Binding Proteins; D000067776:RNA Recognition Motif Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D029701:Saccharomyces cerevisiae Proteins; D016329:Sp1 Transcription Factor; D009713:Nucleotidyltransferases; C020143:FMN adenylyltransferase; D000251:Adenosine Triphosphatases; C481261:Swr1 protein, S cerevisiae; D004265:DNA Helicases; C119652:SRCAP protein, human; C116070:G3BP1 protein, human; D020365:RNA Helicases