Biochemical assay for histone H2A.Z replacement by the yeast SWR1 chromatin remodeling complex

Gaku Mizuguchi, Wei Hua Wu, Samar Alami, Ed Luk

    Research output: Chapter in Book/Report/Conference proceedingChapter

    13 Scopus citations

    Abstract

    The evolutionarily conserved histone variant H2A.Z has an important role in the regulation of gene expression and the establishment of a buffer to the spread of silent heterochromatin. Saccharomyces cerevisiae Swr1, a Swi2/Snf2-related ATPase, is the catalytic core of a multisubunit chromatin remodeling enzyme, called the SWR1 complex, that efficiently replaces conventional histone H2A in nucleosomes with histone H2A.Z. Swr1 is required for the deposition of histone H2A.Z at stereotypical promoter locations in vivo, and Swr1 and H2A.Z commonly regulate a subset of yeast genes. Here, we describe an integrated nucleosome assembly-histone replacement system whereby histone exchange by chromatin remodeling activities may be analyzed in vitro. The system demonstrates ATP- and SWR1-complex-dependent replacement of histone H2A for histone H2A.Z on a preassembled nucleosome array. This system may also be adapted to analyze dynamic interactions between chromatin remodeling and modifying enzymes, histone chaperones, and nucleosome substrates containing canonical, variant, or covalently modified histones.

    Original languageEnglish (US)
    Title of host publicationMethods in Enzymology
    PublisherAcademic Press Inc.
    Pages275-291
    Number of pages17
    DOIs
    StatePublished - 2012

    Publication series

    NameMethods in Enzymology
    Volume512
    ISSN (Print)0076-6879
    ISSN (Electronic)1557-7988

    Keywords

    • Chromatin remodeling
    • H2A.Z
    • Histone exchange
    • Histone octamer
    • Histone variant
    • Htz1
    • Isw1
    • Nap1
    • Nucleosome assembly
    • Swr1

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

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  • Cite this

    Mizuguchi, G., Wu, W. H., Alami, S., & Luk, E. (2012). Biochemical assay for histone H2A.Z replacement by the yeast SWR1 chromatin remodeling complex. In Methods in Enzymology (pp. 275-291). (Methods in Enzymology; Vol. 512). Academic Press Inc.. https://doi.org/10.1016/B978-0-12-391940-3.00012-3