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

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Hannak,  Eva
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Oegema,  Karen
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Kirkham,  Matthew
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Habermann,  Bianca
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Hyman,  Anthony A.
Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Society;

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Citation

Hannak, E., Oegema, K., Kirkham, M., Gönczy, P., Habermann, B., & Hyman, A. A. (2002). The kinetically dominant assembly pathway for centrosomal asters in Caenorhabditis elegans is gamma-tubulin dependent. Journal of Cell Biology, 157(4), 591-602.


Cite as: http://hdl.handle.net/21.11116/0000-0001-13C8-8
Abstract
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 approximately 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.