Purification of a Human SRCAP Complex That Remodels Chromatin by Incorporating the Histone Variant H2A.Z into Nucleosomes

The Snf-2-related CREB-binding protein activator protein (SRCAP) serves as a coactivator for a number of transcription factors known to interact with CBP. Swr1, the closest Saccharomyces cerevisiae ortholog of SRCAP, is a component of the chromatin remodeling complex SWR-C, which catalyzes exchange...

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Bibliographic Details
Published in:Biochemistry (Easton) 2006-05, Vol.45 (17), p.5671-5677
Main Authors: Ruhl, Donald D, Jin, Jingji, Cai, Yong, Swanson, Selene, Florens, Laurence, Washburn, Michael P, Conaway, Ronald C, Conaway, Joan Weliky, Chrivia, John C
Format: Article
Language:English
Subjects:
Adenosine Triphosphatases - isolation & purification
Adenosine Triphosphatases - metabolism
Chromatin - drug effects
Chromatin - metabolism
Chromatography, Ion Exchange - methods
Histones - metabolism
Multiprotein Complexes - isolation & purification
Multiprotein Complexes - metabolism
Nucleosomes - metabolism
Proteomics
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins - metabolism
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Summary:The Snf-2-related CREB-binding protein activator protein (SRCAP) serves as a coactivator for a number of transcription factors known to interact with CBP. Swr1, the closest Saccharomyces cerevisiae ortholog of SRCAP, is a component of the chromatin remodeling complex SWR-C, which catalyzes exchange of the histone variant H2A.Z into nucleosomes. In this report, we use a combination of conventional chromatography and anti-SRCAP immunoaffinity chromatography to purify a native human SRCAP complex with a polypeptide composition similar to that of SWR-C, and we show for the first time that this SRCAP-containing complex supports ATP-dependent exchange of histone dimers containing H2B and H2A.Z into mononucleosomes reconstituted with recombinant H2A, H2B, H3, and H4. Our findings, together with previous evidence implicating H2A.Z in transcriptional regulation, suggest that SRCAP's coactivator function may depend on its ability to promote incorporation of H2A.Z into chromatin.
ISSN:0006-2960
1520-4995
DOI:10.1021/bi060043d