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Published online
doi:10.1083/jcb.200709102
The Journal of Cell Biology, Vol. 180, No. 4, 771-785
The Rockefeller University Press, 0021-9525 $30.00
© Dammermann et al.
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Article

SAS-4 is recruited to a dynamic structure in newly forming centrioles that is stabilized by the {gamma}-tubulin–mediated addition of centriolar microtubules



Alexander Dammermann, Paul S. Maddox, Arshad Desai, and Karen Oegema

Department of Cellular and Molecular Medicine, Ludwig Institute for Cancer Research, University of California, San Diego, La Jolla, CA 92093

Correspondence to A. Dammermann: adammermann{at}ucsd.edu; or K. Oegema: koegema{at}ucsd.edu

Centrioles are surrounded by pericentriolar material (PCM), which is proposed to promote new centriole assembly by concentrating {gamma}-tubulin. Here, we quantitatively monitor new centriole assembly in living Caenorhabditis elegans embryos, focusing on the conserved components SAS-4 and SAS-6. We show that SAS-4 and SAS-6 are coordinately recruited to the site of new centriole assembly and reach their maximum levels during S phase. Centriolar SAS-6 is subsequently reduced by a mechanism intrinsic to the early assembly pathway that does not require progression into mitosis. Centriolar SAS-4 remains in dynamic equilibrium with the cytoplasmic pool until late prophase, when it is stably incorporated in a step that requires {gamma}-tubulin and microtubule assembly. These results indicate that {gamma}-tubulin in the PCM stabilizes the nascent daughter centriole by promoting microtubule addition to its outer wall. Such a mechanism may help restrict new centriole assembly to the vicinity of preexisting parent centrioles that recruit PCM.

P.S. Maddox's present address is Department of Pathology and Cell Biology, Institute for Research in Immunology and Cancer, Faculty of Medicine, Université de Montréal, Pavillon Marcelle-Coutu, Quai 20, Montreal QC H3T 1J4, Canada

Abbreviations used in this paper: DIC, differential interference contrast; dsRNA, double-stranded RNA; HU, hydroxyurea; PCM, pericentriolar material.


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