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Histone H2A.Z acid patch residues required for deposition and function

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Abstract

The incorporation of histone variants is one mechanism used by the eukaryotic cell to alter the generally repressive chromatin template. However, the exact molecular mechanisms that direct this incorporation are not well understood. The SWR1 chromatin remodeling complex that binds to and directs incorporation of histone variant H2A.Z into chromatin has been characterized, but significantly less information is available concerning the requirements on the H2A.Z target molecule. We performed an unbiased mutagenic screen designed to elucidate the function of H2A.Z in Saccharomyces cerevisiae. The screen identified residues within the conserved acidic patch of H2A.Z as being important for the function of the variant. We characterized single point mutations in the patch that are phenotypically sensitive to a variety of growth conditions and are expressed at lower protein levels, but are functionally defective (htz1-D99A, htz1-D99K, and htz1-E101K). The mutants were significantly less detectable by chromatin immunoprecipitation at PHO5, a gene previously described to be enriched for H2A.Z. These results identify acidic patch residues of H2A.Z that are critical for mediating deposition and function in chromatin, and represent potential candidates for the interaction of H2A.Z with its deposition and/or targeting machinery.

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Acknowledgments

We gratefully acknowledge Jinmei Li for plasmid constructions, Peter Decker and Alex Ward for technical advice, and Nazir Barekzi for technical assistance. This work was supported by GM28920 and GM60444 from the National Institutes of General Medical Sciences, National Institutes of Health, to M.M.S. M.S.S. was supported in part by a Visiting Professorship from the American Society for Cell Biology.

Conflict of interest

The authors declare that they have no conflict of interest.

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Author notes

  1. Kurt Jensen

    Present address: Department of Neurology, McKnight Brain Institute, University of Florida, 100 S. Newell Dr., P.O. Box 100236, Gainesville, FL, 32610-0236, USA

  2. Maria Soledad Santisteban

    Present address: Department of Biology, University of North Carolina at Pembroke, One University Drive, P.O. Box 1510, Pembroke, NC, 28372-1510, USA

Authors and Affiliations

  1. Department of Microbiology, University of Virginia Health System, University of Virginia, P.O. Box 800734, Charlottesville, VA, 22908-0734, USA

    Kurt Jensen, Maria Soledad Santisteban, Craig Urekar & M. Mitchell Smith

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  1. Kurt Jensen
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  2. Maria Soledad Santisteban
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Correspondence to Kurt Jensen.

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Communicated by A. Aguilera.

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Jensen, K., Santisteban, M.S., Urekar, C. et al. Histone H2A.Z acid patch residues required for deposition and function. Mol Genet Genomics 285, 287–296 (2011). https://doi.org/10.1007/s00438-011-0604-5

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  • DOI: https://doi.org/10.1007/s00438-011-0604-5

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