title: A method for genetically installing site-specific acetylation in recombinant histones defines the effects of H3 K56 acetylation.
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.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, England, UK.
authors.forename: Heinz
authors.initials: H
authors.lastname: Neumann
authors.forename: Susan M
authors.initials: SM
authors.lastname: Hancock
authors.forename: Ruth
authors.initials: R
authors.lastname: Buning
authors.forename: Andrew
authors.initials: A
authors.lastname: Routh
authors.forename: Lynda
authors.initials: L
authors.lastname: Chapman
authors.forename: Joanna
authors.initials: J
authors.lastname: Somers
authors.forename: Tom
authors.initials: T
authors.lastname: Owen-Hughes
authors.forename: John
authors.initials: J
authors.lastname: van Noort
authors.forename: Daniela
authors.initials: D
authors.lastname: Rhodes
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
mesh_terms: D000107:Acetylation; D019943:Amino Acid Substitution / Q000502:physiology; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics; D042002:Chromatin Assembly and Disassembly / Q000187:drug effects / Q000502:physiology; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006657:Histones / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives / Q000235:genetics / Q000378:metabolism*; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D012965:Sodium Chloride / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D006657:Histones; D009707:Nucleosomes; C103337:RSC complex, S cerevisiae; D011994:Recombinant Proteins; C445179:SWI-SNF-B chromatin-remodeling complex; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C002315:N(alpha)-acetyllysine; D012965:Sodium Chloride; D000604:Amino Acyl-tRNA Synthetases; D008239:Lysine