Purification of a human SRCAP complex that remodels chromatin by incorporating the histone variant H2A.Z into nucleosomes

Donald D. Ruhl, Jingji Jin, Yong Cai, Selene Swanson, Laurence Florens, Michael P. Washburn, Ronald C. Conaway, Joan Weliky Conaway, John C. Chrivia

Research output: Contribution to journalArticlepeer-review

178 Scopus citations

Abstract

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-SRC AP 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.

Original languageEnglish (US)
Pages (from-to)5671-5677
Number of pages7
JournalBiochemistry
Volume45
Issue number17
DOIs
StatePublished - May 2 2006
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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