N terminus of Swr1 binds to histone H2AZ and provides a platform for subunit assembly in the chromatin remodeling complex

Wei Hua Wu, Chwen Huey Wu, Andreas Ladurner, Gaku Mizuguchi, Debbie Wei, Hua Xiao, Ed Luk, Anand Ranjant, Carl Wu

    Research output: Contribution to journalArticle

    58 Citations (Scopus)

    Abstract

    Variant histone H2AZ-containing nucleosomes are involved in the regulation of gene expression. In Saccharomyces cerevisiae, chromatin deposition of histone H2AZ is mediated by the fourteen-subunit SWR1 complex, which catalyzes ATP-dependent exchange of nucleosomal histone H2A for H2AZ. Previous work defined the role of seven SWR1 subunits (Swr1 ATPase, Swc2, Swc3, Arp6, Swc5, Yaf9, and Swc6) in maintaining complex integrity and H2AZ histone replacement activity. Here we examined the function of three additional SWR1 subunits, bromodomain containing Bdf1, actin-related protein Arp4 and Swc7, by analyzing affinity-purified mutant SWR1 complexes. We observed that depletion of Arp4 (arp4-td) substantially impaired the association of Bdf1, Yaf9, and Swc4. In contrast, loss of either Bdf1 or Swc7 had minimal effects on overall complex integrity. Furthermore, the basic H2AZ histone replacement activity of SWR1 in vitro required Arp4, but not Bdf1 or Swc7. Thus, three out of fourteen SWR1 subunits, Bdf1, Swc7, and previously noted Swc3, appear to have roles auxiliary to the basic histone replacement activity. The N-terminal region of the Swr1 ATPase subunit is necessary and sufficient to direct association of Bdf1 and Swc7, as well as Arp4, Act1, Yaf9 and Swc4. This same region contains an additional H2AZ-H2B specific binding site, distinct from the previously identified Swc2 subunit. These findings suggest that one SWR1 enzyme might be capable of binding two H2AZ-H2B dimers, and provide further insight on the hierarchy and interdependency of molecular interactions within the SWR1 complex.

    Original languageEnglish (US)
    Pages (from-to)6200-6207
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume284
    Issue number10
    DOIs
    StatePublished - Mar 6 2009

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    Chromatin Assembly and Disassembly
    Histones
    Chromatin
    Adenosine Triphosphatases
    Molecular interactions
    Nucleosomes
    Gene Expression Regulation
    Gene expression
    Dimers
    Yeast
    Saccharomyces cerevisiae
    Actins
    Adenosine Triphosphate
    Binding Sites
    Enzymes
    Proteins

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    N terminus of Swr1 binds to histone H2AZ and provides a platform for subunit assembly in the chromatin remodeling complex. / Wu, Wei Hua; Wu, Chwen Huey; Ladurner, Andreas; Mizuguchi, Gaku; Wei, Debbie; Xiao, Hua; Luk, Ed; Ranjant, Anand; Wu, Carl.

    In: Journal of Biological Chemistry, Vol. 284, No. 10, 06.03.2009, p. 6200-6207.

    Research output: Contribution to journalArticle

    Wu, WH, Wu, CH, Ladurner, A, Mizuguchi, G, Wei, D, Xiao, H, Luk, E, Ranjant, A & Wu, C 2009, 'N terminus of Swr1 binds to histone H2AZ and provides a platform for subunit assembly in the chromatin remodeling complex', Journal of Biological Chemistry, vol. 284, no. 10, pp. 6200-6207. https://doi.org/10.1074/jbc.M808830200
    Wu, Wei Hua ; Wu, Chwen Huey ; Ladurner, Andreas ; Mizuguchi, Gaku ; Wei, Debbie ; Xiao, Hua ; Luk, Ed ; Ranjant, Anand ; Wu, Carl. / N terminus of Swr1 binds to histone H2AZ and provides a platform for subunit assembly in the chromatin remodeling complex. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 10. pp. 6200-6207.
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    abstract = "Variant histone H2AZ-containing nucleosomes are involved in the regulation of gene expression. In Saccharomyces cerevisiae, chromatin deposition of histone H2AZ is mediated by the fourteen-subunit SWR1 complex, which catalyzes ATP-dependent exchange of nucleosomal histone H2A for H2AZ. Previous work defined the role of seven SWR1 subunits (Swr1 ATPase, Swc2, Swc3, Arp6, Swc5, Yaf9, and Swc6) in maintaining complex integrity and H2AZ histone replacement activity. Here we examined the function of three additional SWR1 subunits, bromodomain containing Bdf1, actin-related protein Arp4 and Swc7, by analyzing affinity-purified mutant SWR1 complexes. We observed that depletion of Arp4 (arp4-td) substantially impaired the association of Bdf1, Yaf9, and Swc4. In contrast, loss of either Bdf1 or Swc7 had minimal effects on overall complex integrity. Furthermore, the basic H2AZ histone replacement activity of SWR1 in vitro required Arp4, but not Bdf1 or Swc7. Thus, three out of fourteen SWR1 subunits, Bdf1, Swc7, and previously noted Swc3, appear to have roles auxiliary to the basic histone replacement activity. The N-terminal region of the Swr1 ATPase subunit is necessary and sufficient to direct association of Bdf1 and Swc7, as well as Arp4, Act1, Yaf9 and Swc4. This same region contains an additional H2AZ-H2B specific binding site, distinct from the previously identified Swc2 subunit. These findings suggest that one SWR1 enzyme might be capable of binding two H2AZ-H2B dimers, and provide further insight on the hierarchy and interdependency of molecular interactions within the SWR1 complex.",
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    AU - Wei, Debbie

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    AU - Luk, Ed

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