2. Academic Validation
  3. The Flexible Ends of CENP-A Nucleosome Are Required for Mitotic Fidelity

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

  • Mol Cell. 2016 Aug 18;63(4):674-685. doi: 10.1016/j.molcel.2016.06.023.
Yohan Roulland 1,  Khalid Ouararhni 2,  Mladen Naidenov 3,  Lorrie Ramos 1,  Muhammad Shuaib 2,  Sajad Hussain Syed 4,  Imtiaz Nizar Lone 4,  Ramachandran Boopathi 4,  Emeline Fontaine 1,  Gabor Papai 5,  Hiroaki Tachiwana 6,  Thierry Gautier 1,  Dimitrios Skoufias 7,  Kiran Padmanabhan 8,  Jan Bednar 1,  Hitoshi Kurumizaka 6,  Patrick Schultz 5,  Dimitar Angelov 9,  Ali Hamiche 10,  Stefan Dimitrov 11
Affiliations

Affiliations

  • 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.
PMID: 27499292 DOI: 10.1016/j.molcel.2016.06.023
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.

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