A method for genetically installing site-specific acetylation in recombinant histories defines the effects of H3 K56 acetylation

Heinz Neumann, Susan M. Hancock, Ruth Buning, Andrew Routh, Lynda Chapman, Joanna Somers, Tom Owen-Hughes, John van Noort, Daniela Rhodes, Jason W. Chin

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    319 Citations (Scopus)

    Abstract

    Lysine acetylation of histones defines the epigenetic status of human embryonic stem cells and orchestrates DNA replication, chromosome condensation, transcription, telomeric silencing, and DNA repair. A detailed mechanistic explanation of these phenomena is impeded by the limited availability of homogeneously acetylated histones. We report a general method for the production of homogeneously and site-specifically acetylated recombinant histones by genetically encoding acetyl-lysine. We reconstitute histone octamers, nucleosomes, and nucleosomal arrays bearing defined acetylated lysine residues. With these designer nucleosomes, we demonstrate that, in contrast to the prevailing dogma, acetylation of H3 K56 does not directly affect the compaction of chromatin and has modest effects on remodeling by SWI/SNF and RSC. Single-molecule FRET experiments reveal that H3 K56 acetylation increases DNA breathing 7-fold. Our results provide a molecular and mechanistic underpinning for cellular phenomena that have been linked with K56 acetylation.

    Original languageEnglish
    Pages (from-to)153-163
    Number of pages11
    JournalMolecular Cell
    Volume36
    Issue number1
    DOIs
    Publication statusPublished - 9 Oct 2009

    Keywords

    • NUCLEOSOME CORE PARTICLE
    • NM CHROMATIN FIBER
    • HISTONE H3
    • LINKER HISTONE
    • SACCHAROMYCES-CEREVISIAE
    • H4-K16 ACETYLATION
    • BINDING MODULES
    • DNA-DAMAGE
    • PHD FINGER
    • LYSINE 56

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