Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants

Junbiao Dai, Edel M. Hyland, Daniel S. Yuan, Hailiang Huang, Joel S. Bader, Jef D. Boeke

    Research output: Contribution to journalArticle

    Abstract

    Nucleosome structural integrity underlies the regulation of DNA metabolism and transcription. Using a synthetic approach, a versatile library of 486 systematic histone H3 and H4 substitution and deletion mutants that probes the contribution of each residue to nucleosome function was generated in Saccharomyces cerevisiae. We probed fitness contributions of each residue to perturbations of chromosome integrity and transcription, mapping global patterns of chemical sensitivities and requirements for transcriptional silencing onto the nucleosome surface. Each histone mutant was tagged with unique molecular barcodes, facilitating identification of histone mutant pools through barcode amplification, labeling, and TAG microarray hybridization. Barcodes were used to score complex phenotypes such as competitive fitness in a chemostat, DNA repair proficiency, and synthetic genetic interactions, revealing new functions for distinct histone residues and new interdependencies among nucleosome components and their modifiers.

    Original languageEnglish (US)
    Pages (from-to)1066-1078
    Number of pages13
    JournalCell
    Volume134
    Issue number6
    DOIs
    StatePublished - Sep 19 2008

    Keywords

    • CHEMBIO
    • PROTEINS

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

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  • Cite this

    Dai, J., Hyland, E. M., Yuan, D. S., Huang, H., Bader, J. S., & Boeke, J. D. (2008). Probing Nucleosome Function: A Highly Versatile Library of Synthetic Histone H3 and H4 Mutants. Cell, 134(6), 1066-1078. https://doi.org/10.1016/j.cell.2008.07.019