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Published 13 May 2002. doi:10.1083/jcb.200202047
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© The Rockefeller University Press, 0021-9525/2002/5/591 $5.00
The Journal of Cell Biology, Volume 157, Number 4, May 13, 2002 591-602

Article

 The kinetically dominant assembly pathway for centrosomal asters in Caenorhabditis elegans is {gamma}-tubulin dependent

Eva Hannak1, Karen Oegema1, Matthew Kirkham1, Pierre Gönczy2, Bianca Habermann1 and Anthony A. Hyman1

1 Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
2 Institut Suisse de Recherche Expérimentale sur le Cancer, CH-1066 Epalinges s/Lausanne, Switzerland

Address correspondence to Anthony Hyman, Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany. Tel.: 49-351-210-1700. Fax: 49-351-210-1289. E-mail: hyman{at}mpi-cbg.de

{gamma}-Tubulin–containing complexes are thought to nucleate and anchor centrosomal microtubules (MTs). Surprisingly, a recent study (Strome, S., J. Powers, M. Dunn, K. Reese, C.J. Malone, J. White, G. Seydoux, and W. Saxton. Mol. Biol. Cell. 12:1751–1764) showed that centrosomal asters form in Caenorhabditis elegans embryos depleted of {gamma}-tubulin by RNA-mediated interference (RNAi). Here, we investigate the nucleation and organization of centrosomal MT asters in C. elegans embryos severely compromised for {gamma}-tubulin function. We characterize embryos depleted of ~98% centrosomal {gamma}-tubulin by RNAi, embryos expressing a mutant form of {gamma}-tubulin, and embryos depleted of a {gamma}-tubulin–associated protein, CeGrip-1. In all cases, centrosomal asters fail to form during interphase but assemble as embryos enter mitosis. The formation of these mitotic asters does not require ZYG-9, a centrosomal MT-associated protein, or cytoplasmic dynein, a minus end–directed motor that contributes to self-organization of mitotic asters in other organisms. By kinetically monitoring MT regrowth from cold-treated mitotic centrosomes in vivo, we show that centrosomal nucleating activity is severely compromised by {gamma}-tubulin depletion. Thus, although unknown mechanisms can support partial assembly of mitotic centrosomal asters, {gamma}-tubulin is the kinetically dominant centrosomal MT nucleator.

Key Words: microtubule; mitosis; grip; Spc; tbg-1


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