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

doi:10.1016/j.molcel.2009.07.027

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

Research output: Contribution to journal › Article › peer-review

420 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 language	English
Pages (from-to)	153-163
Number of pages	11
Journal	Molecular Cell
Volume	36
Issue number	1
DOIs	https://doi.org/10.1016/j.molcel.2009.07.027
Publication status	Published - 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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title = "A method for genetically installing site-specific acetylation in recombinant histories 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.",

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

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AU - Neumann, Heinz

AU - Hancock, Susan M.

AU - Buning, Ruth

AU - Routh, Andrew

AU - Chapman, Lynda

AU - Somers, Joanna

AU - Owen-Hughes, Tom

AU - van Noort, John

AU - Rhodes, Daniela

AU - Chin, Jason W.

PY - 2009/10/9

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N2 - 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.

AB - 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.

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KW - NM CHROMATIN FIBER

KW - HISTONE H3

KW - LINKER HISTONE

KW - SACCHAROMYCES-CEREVISIAE

KW - H4-K16 ACETYLATION

KW - BINDING MODULES

KW - DNA-DAMAGE

KW - PHD FINGER

KW - LYSINE 56

U2 - 10.1016/j.molcel.2009.07.027

DO - 10.1016/j.molcel.2009.07.027

M3 - Article

C2 - 19818718

SN - 1097-2765

VL - 36

SP - 153

EP - 163

JO - Molecular Cell

JF - Molecular Cell

IS - 1

ER -

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

Abstract

Keywords

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