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Originally published as MBC in Press, 10.1091/mbc.E06-07-0604 on July 11, 2007

Vol. 18, Issue 9, 3667-3680, September 2007

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Chromatin Remodeling Proteins Interact with Pericentrin to Regulate Centrosome IntegrityFormula Formula

James Edward Sillibourne*, Bénédicte Delaval, Sambra Redick, Manisha Sinha, and Stephen John Doxsey

Program in Molecular Medicine, University of Massachusetts, Worcester, MA 01605

Submitted July 17, 2006; Revised June 12, 2007; Accepted June 29, 2007
Monitoring Editor: Tim Stearns

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.

This article was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E06-07-0604) on July 11, 2007.

Formula Formula The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

* Present address: Institut Curie, Section Recherche Unité Mixte de Recherche 144 du Centre National de la Recherche Scientifique, 26 Rue d'Ulm, Paris 75248, France.

Address correspondence to: Stephen J. Doxsey (stephen.doxsey{at}umassmed.edu).




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