Insights into the molecular architecture and histone H3-H4 deposition mechanism of yeast Chromatin assembly factor 1
- European Molecular Biology Laboratory, France
- CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, France
- Institut Curie, France
- Université Grenoble Alpes, France
- CEA, CNRS, Université Paris-Sud, France
- Deutsches Krebsforschungszentrum, Germany
Abstract
How the very first step in nucleosome assembly, deposition of histone H3-H4 as tetramers or dimers on DNA, is accomplished remains largely unclear. Here we report that yeast chromatin assembly factor 1 (CAF1), a conserved histone chaperone complex that deposits H3-H4 during DNA replication, binds a single H3-H4 heterodimer in solution. We identify a new DNA binding domain in the large Cac1 subunit of CAF1, which is required for high-affinity DNA binding by the CAF1 three-subunit complex, and which is distinct from the previously described C-terminal winged-helix domain. CAF1 binds preferentially to DNA molecules longer than 40 bp, and two CAF1-H3-H4 complexes concertedly associate with DNA molecules of this size, resulting in deposition of H3-H4 tetramers. While DNA binding is not essential for H3-H4 tetrasome deposition in vitro, it is required for efficient DNA synthesis-coupled nucleosome assembly. Mutant histones with impaired H3-H4 tetramerization interactions fail to release from CAF1, indicating that DNA deposition of H3-H4 tetramers by CAF1 requires a hierarchical cooperation between DNA binding, H3-H4 deposition and histone tetramerization.
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Funding
Agence Nationale de la Recherche (ANR-16-CE11-0028-02)
- Paul Victor Sauer
- Jennifer Timm
- Danni Liu
- David Sitbon
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2017, Sauer et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Insights into the molecular architecture and histone H3-H4 deposition mechanism of yeast Chromatin assembly factor 1
eLife 6:e23474.
https://doi.org/10.7554/eLife.23474
