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Published online 6 February 2006. doi:10.1083/jcb.200510028
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 172, Number 4, 505-515
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Article

 NEDD1-dependent recruitment of the {gamma}-tubulin ring complex to the centrosome is necessary for centriole duplication and spindle assembly

Laurence Haren1, Marie-Hélène Remy1, Ingrid Bazin1, Isabelle Callebaut2, Michel Wright1, and Andreas Merdes1

1 Institut de Sciences et Technologies du Médicament de Toulouse, Centre National de la Recherche Scientifique/Pierre Fabre, 31400 Toulouse, France
2 Institut de Minéralogie et de Physique des Milieux Condensés, Centre National de la Recherche Scientifique, 75252 Paris, Cedex 05, France

Correspondence to Andreas Merdes: andreas.merdes{at}istmt.cnrs.fr

The centrosome is the major microtubule organizing structure in somatic cells. Centrosomal microtubule nucleation depends on the protein {gamma}-tubulin. In mammals, {gamma}-tubulin associates with additional proteins into a large complex, the {gamma}-tubulin ring complex ({gamma}TuRC). We characterize NEDD1, a centrosomal protein that associates with {gamma}TuRCs. We show that the majority of {gamma}TuRCs assemble even after NEDD1 depletion but require NEDD1 for centrosomal targeting. In contrast, NEDD1 can target to the centrosome in the absence of {gamma}-tubulin. NEDD1-depleted cells show defects in centrosomal microtubule nucleation and form aberrant mitotic spindles with poorly separated poles. Similar spindle defects are obtained by overexpression of a fusion protein of GFP tagged to the carboxy-terminal half of NEDD1, which mediates binding to {gamma}TuRCs. Further, we show that depletion of NEDD1 inhibits centriole duplication, as does depletion of {gamma}-tubulin. Our data suggest that centriole duplication requires NEDD1-dependent recruitment of {gamma}-tubulin to the centrosome.

L. Haren and M.-H. Remy contributed equally to this paper.

Abbreviations used in this paper: {gamma}TuRC, {gamma}-tubulin ring complex; {gamma}TuSC, {gamma}-tubulin small complex; GCP, {gamma}-complex protein; NuMA, nuclear mitotic apparatus protein; RNAi, RNA interference; siRNA, small interfering RNA.


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