title: The Flexible Ends of CENP-A Nucleosome Are Required for Mitotic Fidelity.
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.
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
authors: Yohan
authors: Y
authors: Roulland
authors: 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.
authors: Khalid
authors: K
authors: Ouararhni
authors: 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.
authors: Mladen
authors: M
authors: Naidenov
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
authors: Lorrie
authors: L
authors: Ramos
authors: 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.
authors: Muhammad
authors: M
authors: Shuaib
authors: 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.
authors: Sajad Hussain
authors: SH
authors: Syed
authors: 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.
authors: Imtiaz Nizar
authors: IN
authors: Lone
authors: 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.
authors: Ramachandran
authors: R
authors: Boopathi
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
authors: Emeline
authors: E
authors: Fontaine
authors: Department of Integrated Structural Biology, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC)/Université de Strasbourg/CNRS/INSERM, 67404 Illkirch Cedex, France.
authors: Gabor
authors: G
authors: Papai
authors: Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo 162-8480, Japan.
authors: Hiroaki
authors: H
authors: Tachiwana
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
authors: Thierry
authors: T
authors: Gautier
authors: Institut de Biologie Structurale, Université de Grenoble Alpes, 38044 Grenoble, France.
authors: Dimitrios
authors: D
authors: Skoufias
authors: 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.
authors: Kiran
authors: K
authors: Padmanabhan
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France.
authors: Jan
authors: J
authors: Bednar
authors: Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo 162-8480, Japan.
authors: Hitoshi
authors: H
authors: Kurumizaka
authors: Department of Integrated Structural Biology, Institut de Génétique et Biologie Moléculaire et Cellulaire (IGBMC)/Université de Strasbourg/CNRS/INSERM, 67404 Illkirch Cedex, France.
authors: Patrick
authors: P
authors: Schultz
authors: 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.
authors: Dimitar
authors: D
authors: Angelov
authors: 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.
authors: Ali
authors: A
authors: Hamiche
authors: Université de Grenoble Alpes/INSERM U1209/CNRS UMR 5309, 38042 Grenoble Cedex 9, France. Electronic address: stefan.dimitrov@univ-grenoble-alpes.fr.
authors: Stefan
authors: S
authors: Dimitrov
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D001665:Binding Sites; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000172:deficiency / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D020285:Cryoelectron Microscopy; D048749:Cytokinesis; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D005838:Genotype; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D008938:Mitosis / Q000502:physiology*; D008958:Models, Molecular; D009154:Mutation; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000378:metabolism* / Q000648:ultrastructure; D010641:Phenotype; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; C000619701:Cenpa protein, mouse; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D009707:Nucleosomes; C060905:centromere protein C; D004247:DNA