title: Chromatin remodeling proteins interact with pericentrin to regulate centrosome integrity.
abstract: Pericentrin is an integral centrosomal component that anchors regulatory and structural molecules to centrosomes. In a yeast two-hybrid screen with pericentrin we identified chromodomain helicase DNA-binding protein 4 (CHD4/Mi2beta). CHD4 is part of the multiprotein nucleosome remodeling deacetylase (NuRD) complex. We show that many NuRD components interacted with pericentrin by coimmunoprecipitation and that they localized to centrosomes and midbodies. Overexpression of the pericentrin-binding domain of CHD4 or another family member (CHD3) dissociated pericentrin from centrosomes. Depletion of CHD3, but not CHD4, by RNA interference dissociated pericentrin and gamma-tubulin from centrosomes. Microtubule nucleation/organization, cell morphology, and nuclear centration were disrupted in CHD3-depleted cells. Spindles were disorganized, the majority showing a prometaphase-like configuration. Time-lapse imaging revealed mitotic failure before chromosome segregation and cytokinesis failure. We conclude that pericentrin forms complexes with CHD3 and CHD4, but a distinct CHD3-pericentrin complex is required for centrosomal anchoring of pericentrin/gamma-tubulin and for centrosome integrity.
authors.affiliation: Program in Molecular Medicine, University of Massachusetts, Worcester, MA 01605, USA.
authors.forename: James Edward
authors.initials: JE
authors.lastname: Sillibourne
authors.forename: Bénédicte
authors.initials: B
authors.lastname: Delaval
authors.forename: Sambra
authors.initials: S
authors.lastname: Redick
authors.forename: Manisha
authors.initials: M
authors.lastname: Sinha
authors.forename: Stephen John
authors.initials: SJ
authors.lastname: Doxsey
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D000818:Animals; D000941:Antigens / Q000378:metabolism*; D001324:Autoantigens / Q000737:chemistry / Q000378:metabolism*; D019556:COS Cells; D018385:Centrosome / Q000378:metabolism*; D002522:Chlorocebus aethiops; D042002:Chromatin Assembly and Disassembly; D048749:Cytokinesis; D004265:DNA Helicases / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism*; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D011485:Protein Binding; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D000941:Antigens; D001324:Autoantigens; C115323:CHD4 protein, human; D034741:RNA, Small Interfering; C086178:pericentrin; D006655:Histone Deacetylases; D056245:Mi-2 Nucleosome Remodeling and Deacetylase Complex; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; C115322:CHD3 protein, human