1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics
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Insights into the molecular architecture and histone H3-H4 deposition mechanism of yeast Chromatin assembly factor 1

  1. Paul Victor Sauer
  2. Jennifer Timm
  3. Danni Liu
  4. David Sitbon
  5. Elisabetta Boeri-Erba
  6. Christophe Velours
  7. Norbert Mücke
  8. Jörg Langowski
  9. Françoise Ochsenbein
  10. Geneviève Almouzni
  11. Daniel Panne  Is a corresponding author
  1. European Molecular Biology Laboratory, France
  2. CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, France
  3. Institut Curie, France
  4. Université Grenoble Alpes, France
  5. CEA, CNRS, Université Paris-Sud, France
  6. Deutsches Krebsforschungszentrum, Germany
Research Article
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Cite this article as: eLife 2017;6:e23474 doi: 10.7554/eLife.23474

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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Author details

  1. Paul Victor Sauer

    European Molecular Biology Laboratory, Grenoble, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7204-5863
  2. Jennifer Timm

    European Molecular Biology Laboratory, Grenoble, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Danni Liu

    CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, Gif-sur-Yvette, France
    Competing interests
    The authors declare that no competing interests exist.
  4. David Sitbon

    Institut Curie, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Elisabetta Boeri-Erba

    Institut de Biologie Structurale, Université Grenoble Alpes, Grenoble, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Christophe Velours

    Institute for Integrative Biology of the Cell, CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Norbert Mücke

    Abteilung Biophysik der Makromoleküle, Deutsches Krebsforschungszentrum, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Jörg Langowski

    Abteilung Biophysik der Makromoleküle, Deutsches Krebsforschungszentrum, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. Françoise Ochsenbein

    CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, Gif-sur-Yvette, France
    Competing interests
    The authors declare that no competing interests exist.
  10. Geneviève Almouzni

    Institut Curie, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  11. Daniel Panne

    European Molecular Biology Laboratory, Grenoble, France
    For correspondence
    panne@embl.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9158-5507

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.

Reviewing Editor

  1. Paul D. Kaufman, U. Massachusetts Medical School, United States

Publication history

  1. Received: November 21, 2016
  2. Accepted: March 9, 2017
  3. Accepted Manuscript published: March 18, 2017 (version 1)
  4. Version of Record published: April 25, 2017 (version 2)

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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