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Mol Cell. 2016 Aug 18;63(4):674-685. doi: 10.1016/j.molcel.2016.06.023. Epub 2016 Aug 4.

The Flexible Ends of CENP-A Nucleosome Are Required for Mitotic Fidelity.

Author information

1
Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
2
Department of Functional Genomics and Cancer, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC), Equipe Labellisée Ligue Contre le Cancer/Université de Strasbourg/CNRS/INSERM, 67404 Illkirch Cedex, France.
3
Laboratoire de Biologie Moléculaire de la Cellule (LBMC) CNRS/ENSL/UCBL UMR 5239 and Institut NeuroMyoGène - INMG CNRS/UCBL UMR 5310, Université de Lyon, Ecole Normale Supérieure de Lyon, 69007 Lyon, France.
4
Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France; Laboratoire de Biologie Moléculaire de la Cellule (LBMC) CNRS/ENSL/UCBL UMR 5239 and Institut NeuroMyoGène - INMG CNRS/UCBL UMR 5310, Université de Lyon, Ecole Normale Supérieure de Lyon, 69007 Lyon, France.
5
Department of Integrated Structural Biology, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC)/Université de Strasbourg/CNRS/INSERM, 67404 Illkirch Cedex, France.
6
Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo 162-8480, Japan.
7
Institut de Biologie Structurale, Université de Grenoble Alpes, 38044 Grenoble, France.
8
Institut de Génomique Fonctionnelle de Lyon, Ecole Normale Supérieure de Lyon, Centre National de la Recherche Scientifique, UMR 5242, 46 Allée d'Italie, 69364 Lyon Cedex 07, France.
9
Laboratoire de Biologie Moléculaire de la Cellule (LBMC) CNRS/ENSL/UCBL UMR 5239 and Institut NeuroMyoGène - INMG CNRS/UCBL UMR 5310, Université de Lyon, Ecole Normale Supérieure de Lyon, 69007 Lyon, France. Electronic address: dimitar.anguelov@ens-lyon.fr.
10
Department of Functional Genomics and Cancer, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC), Equipe Labellisée Ligue Contre le Cancer/Université de Strasbourg/CNRS/INSERM, 67404 Illkirch Cedex, France; Qatar Biomedical Research Institute, Hamad Bin Khalifa University, P.O. Box 5825, Qatar Foundation, Doha, Qatar. Electronic address: hamiche@igbmc.fr.
11
Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France. Electronic address: stefan.dimitrov@univ-grenoble-alpes.fr.

Abstract

CENP-A is a histone variant, which replaces histone H3 at centromeres and confers unique properties to centromeric chromatin. The crystal structure of CENP-A nucleosome suggests flexible nucleosomal DNA ends, but their dynamics in solution remains elusive and their implication in centromere function is unknown. Using electron cryo-microscopy, we determined the dynamic solution properties of the CENP-A nucleosome. Our biochemical, proteomic, and genetic data reveal that higher flexibility of DNA ends impairs histone H1 binding to the CENP-A nucleosome. Substituting the 2-turn αN-helix of CENP-A with the 3-turn αN-helix of H3 results in compact particles with rigidified DNA ends, able to bind histone H1. In vivo replacement of CENP-A with H3-CENP-A hybrid nucleosomes leads to H1 recruitment, delocalization of kinetochore proteins, and significant mitotic and cytokinesis defects. Our data reveal that the evolutionarily conserved flexible ends of the CENP-A nucleosomes are essential to ensure the fidelity of the mitotic pathway.

PMID:
27499292
DOI:
10.1016/j.molcel.2016.06.023
[Indexed for MEDLINE]
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