A Method for Genetically Installing Site-Specific Acetylation in Recombinant Histones 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

Research output: Contribution to journalArticle

310 Scopus citations

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 (US)
Pages (from-to)153-163
Number of pages11
JournalMolecular cell
Volume36
Issue number1
DOIs
StatePublished - Oct 9 2009
Externally publishedYes

Keywords

  • DNA
  • PROTEINS

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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    Neumann, H., Hancock, S. M., Buning, R., Routh, A., Chapman, L., Somers, J., Owen-Hughes, T., van Noort, J., Rhodes, D., & Chin, J. W. (2009). A Method for Genetically Installing Site-Specific Acetylation in Recombinant Histones Defines the Effects of H3 K56 Acetylation. Molecular cell, 36(1), 153-163. https://doi.org/10.1016/j.molcel.2009.07.027