Insights into the molecular architecture and histone H3-H4 deposition mechanism of yeast Chromatin assembly factor 1.
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.
European Molecular Biology Laboratory, Grenoble, France.
Paul Victor
0000-0001-7204-5863
PV
Sauer
European Molecular Biology Laboratory, Grenoble, France.
Jennifer
J
Timm
CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, Gif-sur-Yvette, France.|Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France.
Danni
D
Liu
Institut Curie, PSL Research University, CNRS, UMR3664, Equipe Labellisée Ligue contre le Cancer, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, CNRS, UMR3664, Paris, France.
David
D
Sitbon
Université Grenoble Alpes, Institut de Biologie Structurale (IBS), Grenoble, France.|Commissariat à l'Energie Atomique et aux Energies Alternatives (CEA), Grenoble, France.
Elisabetta
E
Boeri-Erba
Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France.
Christophe
C
Velours
Abteilung Biophysik der Makromoleküle, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Norbert
N
Mücke
Abteilung Biophysik der Makromoleküle, Deutsches Krebsforschungszentrum, Heidelberg, Germany.
Jörg
J
Langowski
CEA, DRF, SB2SM, Laboratoire de Biologie Structurale et Radiobiologie, Gif-sur-Yvette, France.|Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Sud, Gif-sur-Yvette, France.
Françoise
F
Ochsenbein
Institut Curie, PSL Research University, CNRS, UMR3664, Equipe Labellisée Ligue contre le Cancer, Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, CNRS, UMR3664, Paris, France.
Geneviève
G
Almouzni
European Molecular Biology Laboratory, Grenoble, France.
Daniel
0000-0001-9158-5507
D
Panne
D004261:DNA Replication; D004271:DNA, Fungal / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D011485:Protein Binding; D055503:Protein Multimerization; D012260:Ribonucleases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
D004271:DNA, Fungal; D006657:Histones; D029701:Saccharomyces cerevisiae Proteins; D012260:Ribonucleases; C079009:POP2 protein, S cerevisiae
DNA replication; S. cerevisiae; biochemistry; biophysics; chromatin; chromatin assembly factor 1; histone chaperone; histones; structural biology