Yeast CAF-1 assembles histone (H3-H4)2 tetramers prior to DNA deposition

Duane D. Winkler; Hui Zhou; Mohd A. Dar; Zhiguo Zhang; Karolin Luger

doi:10.1093/nar/gks812

Yeast CAF-1 assembles histone (H3-H4)₂ tetramers prior to DNA deposition

Duane D. Winkler, Hui Zhou, Mohd A. Dar, Zhiguo Zhang, Karolin Luger

Biochemistry and Molecular Biology

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

56 Scopus citations

Abstract

Following acetylation, newly synthesized H3-H4 is directly transferred from the histone chaperone anti-silencing factor 1 (Asf1) to chromatin assembly factor 1 (CAF-1), another histone chaperone that is critical for the deposition of H3-H4 onto replicating DNA. However, it is unknown how CAF-1 binds and delivers H3-H4 to the DNA. Here, we show that CAF-1 binds recombinant H3-H4 with 10- to 20-fold higher affinity than H2A-H2B in vitro, and H3K56Ac increases the binding affinity of CAF-1 toward H3-H4 2-fold. These results provide a quantitative thermodynamic explanation for the specific H3-H4 histone chaperone activity of CAF-1. Surprisingly, H3-H4 exists as a dimer rather than as a canonical tetramer at mid-to-low nanomolar concentrations. A single CAF-1 molecule binds a cross-linked (H3-H4)₂ tetramer, or two H3-H4 dimers that contain mutations at the (H3-H4)₂ tetramerization interface. These results suggest that CAF-1 binds to two H3-H4 dimers in a manner that promotes formation of a (H3-H4)₂ tetramer. Consistent with this idea, we confirm that CAF-1 synchronously binds two H3-H4 dimers derived from two different histone genes in vivo. Together, the data illustrate a clear mechanism for CAF-1- associated H3-H4 chaperone activity in the context of de novo nucleosome (re)assembly following DNA replication.

Original language	English (US)
Pages (from-to)	10139-10149
Number of pages	11
Journal	Nucleic acids research
Volume	40
Issue number	20
DOIs	https://doi.org/10.1093/nar/gks812
State	Published - Nov 2012

ASJC Scopus subject areas

Genetics

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@article{552ba96ee388440a8f5716dd232412a1,

title = "Yeast CAF-1 assembles histone (H3-H4)2 tetramers prior to DNA deposition",

abstract = "Following acetylation, newly synthesized H3-H4 is directly transferred from the histone chaperone anti-silencing factor 1 (Asf1) to chromatin assembly factor 1 (CAF-1), another histone chaperone that is critical for the deposition of H3-H4 onto replicating DNA. However, it is unknown how CAF-1 binds and delivers H3-H4 to the DNA. Here, we show that CAF-1 binds recombinant H3-H4 with 10- to 20-fold higher affinity than H2A-H2B in vitro, and H3K56Ac increases the binding affinity of CAF-1 toward H3-H4 2-fold. These results provide a quantitative thermodynamic explanation for the specific H3-H4 histone chaperone activity of CAF-1. Surprisingly, H3-H4 exists as a dimer rather than as a canonical tetramer at mid-to-low nanomolar concentrations. A single CAF-1 molecule binds a cross-linked (H3-H4)2 tetramer, or two H3-H4 dimers that contain mutations at the (H3-H4)2 tetramerization interface. These results suggest that CAF-1 binds to two H3-H4 dimers in a manner that promotes formation of a (H3-H4)2 tetramer. Consistent with this idea, we confirm that CAF-1 synchronously binds two H3-H4 dimers derived from two different histone genes in vivo. Together, the data illustrate a clear mechanism for CAF-1- associated H3-H4 chaperone activity in the context of de novo nucleosome (re)assembly following DNA replication.",

author = "Winkler, {Duane D.} and Hui Zhou and Dar, {Mohd A.} and Zhiguo Zhang and Karolin Luger",

note = "Funding Information: Funding for open access charge: The National Institutes of Health [GM-067777 to K.L., F32GM096531 to D.D.W. and GM72719 to Z.Z.]; Howard Hughes Medical Institute (to K.L.).",

year = "2012",

month = nov,

doi = "10.1093/nar/gks812",

language = "English (US)",

volume = "40",

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T1 - Yeast CAF-1 assembles histone (H3-H4)2 tetramers prior to DNA deposition

AU - Winkler, Duane D.

AU - Zhou, Hui

AU - Dar, Mohd A.

AU - Zhang, Zhiguo

AU - Luger, Karolin

N1 - Funding Information: Funding for open access charge: The National Institutes of Health [GM-067777 to K.L., F32GM096531 to D.D.W. and GM72719 to Z.Z.]; Howard Hughes Medical Institute (to K.L.).

PY - 2012/11

Y1 - 2012/11

N2 - Following acetylation, newly synthesized H3-H4 is directly transferred from the histone chaperone anti-silencing factor 1 (Asf1) to chromatin assembly factor 1 (CAF-1), another histone chaperone that is critical for the deposition of H3-H4 onto replicating DNA. However, it is unknown how CAF-1 binds and delivers H3-H4 to the DNA. Here, we show that CAF-1 binds recombinant H3-H4 with 10- to 20-fold higher affinity than H2A-H2B in vitro, and H3K56Ac increases the binding affinity of CAF-1 toward H3-H4 2-fold. These results provide a quantitative thermodynamic explanation for the specific H3-H4 histone chaperone activity of CAF-1. Surprisingly, H3-H4 exists as a dimer rather than as a canonical tetramer at mid-to-low nanomolar concentrations. A single CAF-1 molecule binds a cross-linked (H3-H4)2 tetramer, or two H3-H4 dimers that contain mutations at the (H3-H4)2 tetramerization interface. These results suggest that CAF-1 binds to two H3-H4 dimers in a manner that promotes formation of a (H3-H4)2 tetramer. Consistent with this idea, we confirm that CAF-1 synchronously binds two H3-H4 dimers derived from two different histone genes in vivo. Together, the data illustrate a clear mechanism for CAF-1- associated H3-H4 chaperone activity in the context of de novo nucleosome (re)assembly following DNA replication.

AB - Following acetylation, newly synthesized H3-H4 is directly transferred from the histone chaperone anti-silencing factor 1 (Asf1) to chromatin assembly factor 1 (CAF-1), another histone chaperone that is critical for the deposition of H3-H4 onto replicating DNA. However, it is unknown how CAF-1 binds and delivers H3-H4 to the DNA. Here, we show that CAF-1 binds recombinant H3-H4 with 10- to 20-fold higher affinity than H2A-H2B in vitro, and H3K56Ac increases the binding affinity of CAF-1 toward H3-H4 2-fold. These results provide a quantitative thermodynamic explanation for the specific H3-H4 histone chaperone activity of CAF-1. Surprisingly, H3-H4 exists as a dimer rather than as a canonical tetramer at mid-to-low nanomolar concentrations. A single CAF-1 molecule binds a cross-linked (H3-H4)2 tetramer, or two H3-H4 dimers that contain mutations at the (H3-H4)2 tetramerization interface. These results suggest that CAF-1 binds to two H3-H4 dimers in a manner that promotes formation of a (H3-H4)2 tetramer. Consistent with this idea, we confirm that CAF-1 synchronously binds two H3-H4 dimers derived from two different histone genes in vivo. Together, the data illustrate a clear mechanism for CAF-1- associated H3-H4 chaperone activity in the context of de novo nucleosome (re)assembly following DNA replication.

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DO - 10.1093/nar/gks812

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EP - 10149

JO - Nucleic acids research

JF - Nucleic acids research

IS - 20

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Yeast CAF-1 assembles histone (H3-H4)₂ tetramers prior to DNA deposition

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