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Published 13 February 2006. doi:10.1083/jcb.200511071
The Rockefeller University Press, 0021-9525 $8.00
JCB, Volume 172, Number 4, 517-528
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Article

Drosophila melanogaster {gamma}-TuRC is dispensable for targeting {gamma}-tubulin to the centrosome and microtubule nucleation



Christel Vérollet1, Nathalie Colombié1, Thomas Daubon1, Henri-Marc Bourbon2, Michel Wright1, and Brigitte Raynaud-Messina1

1 Centre de Recherche en Pharmacologie, Santé, UMR 2587, Centre National de la Recherche Scientifique-Pierre Fabre, Institut de Sciences et Technologies du Médicament de Toulouse, 31432 Toulouse, Cedex 4, France
2 Centre de Biologie du Développement, UMR 5547, Centre National de la Recherche Scientifique-Université Paul Sabatier, IFR109, 31062 Toulouse, Cedex 4, France

Correspondence to Brigitte Raynaud-Messina: brigitte.raynaud-messina{at}istmt.cnrs.fr

In metazoans, {gamma}-tubulin acts within two main complexes, {gamma}-tubulin small complexes ({gamma}-TuSCs) and {gamma}-tubulin ring complexes ({gamma}-TuRCs). In higher eukaryotes, it is assumed that microtubule nucleation at the centrosome depends on {gamma}-TuRCs, but the role of {gamma}-TuRC components remains undefined.

For the first time, we analyzed the function of all four {gamma}-TuRC–specific subunits in Drosophila melanogaster: Dgrip75, Dgrip128, Dgrip163, and Dgp71WD. Grip-motif proteins, but not Dgp71WD, appear to be required for {gamma}-TuRC assembly. Individual depletion of {gamma}-TuRC components, in cultured cells and in vivo, induces mitotic delay and abnormal spindles. Surprisingly, {gamma}-TuSCs are recruited to the centrosomes. These defects are less severe than those resulting from the inhibition of {gamma}-TuSC components and do not appear critical for viability. Simultaneous cosilencing of all {gamma}-TuRC proteins leads to stronger phenotypes and partial recruitment of {gamma}-TuSC. In conclusion, {gamma}-TuRCs are required for assembly of fully functional spindles, but we suggest that {gamma}-TuSC could be targeted to the centrosomes, which is where basic microtubule assembly activities are maintained.

Abbreviations used in this paper: Cnn, centrosomin; dsRNA, double-stranded RNA; {gamma}-TuRC, {gamma}-tubulin ring complex; {gamma}-TuSC, {gamma}-tubulin small complex; P95, probability of 95%; RNAi, RNA interference.


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