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Swc2 is a widely conserved H2AZ-binding module essential for ATP-dependent histone exchange

Nature Structural & Molecular Biology volume 12, pages 10641071 (2005) | Download Citation

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Abstract

The histone variant H2AZ is incorporated preferentially at specific locations in chromatin to modulate chromosome functions. In Saccharomyces cerevisiae, deposition of histone H2AZ is mediated by the multiprotein SWR1 complex, which catalyzes ATP-dependent exchange of nucleosomal histone H2A for H2AZ. Here, we define interactions between SWR1 components and H2AZ, revealing a link between the ATPase domain of Swr1 and three subunits required for the binding of H2AZ. We discovered that Swc2 binds directly to and is essential for transfer of H2AZ. Swc6 and Arp6 are necessary for the association of Swc2 and for nucleosome binding, whereas other subunits, Swc5 and Yaf9, are required for H2AZ transfer but neither H2AZ nor nucleosome binding. Finally, the C-terminal α-helix of H2AZ is crucial for its recognition by SWR1. These findings provide insight on the initial events of histone exchange.

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Acknowledgements

We thank X. Shen (University of Texas, M.D. Anderson Cancer Center) for the pHtz1-2Flag plasmid, T. Tsukiyama (Fred Hutchinson Cancer Research Center) for the yeast strain W1544-4C and the 3× Flag–tagging plasmid and J. Landry and H. Xiao for helpful discussions. This work was supported by the Intramural Research Program of the US National Cancer Institute.

Author information

Affiliations

  1. Laboratory of Molecular Cell Biology, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Building 37, Room 6068, Bethesda, Maryland 20892-4255, USA.

    • Wei-Hua Wu
    • , Samar Alami
    • , Edward Luk
    • , Chwen-Huey Wu
    • , Subhojit Sen
    • , Gaku Mizuguchi
    • , Debbie Wei
    •  & Carl Wu

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Carl Wu.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    mRNA expression levels of SWC2, ARP6, SWC6 and HTZ1 in wild-type, swc2delta arp6Δ and swc6Δ mutant strains.

  2. 2.

    Supplementary Fig. 2

    Protein sequence alignment of S. cerevisiae Swc2 and related proteins from S. pombe, D. melanogaster, M. musculus and H. sapiens.

  3. 3.

    Supplementary Fig. 3

    Association of multiple subunits with Swr1 N2 and ATPase/Insert domains are required for functional replacement of H2AZ in vitro.

  4. 4.

    Supplementary Fig. 4

    An alignment of D. melanogaster H2AvD and S. cerevisiae Htz1 and H2A.

  5. 5.

    Supplementary Fig. 5

    Purified bacterially expressed His6-tagged proteins used in protein interaction assays.

  6. 6.

    Supplementary Table 1

    Yeast strains used in this study.

  7. 7.

    Supplementary Methods

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DOI

https://doi.org/10.1038/nsmb1023

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