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

Research output: Contribution to journalArticle

53 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 languageEnglish (US)
Pages (from-to)10139-10149
Number of pages11
JournalNucleic acids research
Volume40
Issue number20
DOIs
StatePublished - Nov 2012

ASJC Scopus subject areas

  • Genetics

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    Winkler, D. D., Zhou, H., Dar, M. A., Zhang, Z., & Luger, K. (2012). Yeast CAF-1 assembles histone (H3-H4)2 tetramers prior to DNA deposition. Nucleic acids research, 40(20), 10139-10149. https://doi.org/10.1093/nar/gks812