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Vol. 17, Issue 1, 272-282, January 2006

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The Drosophila -Tubulin Small Complex Subunit Dgrip84 Is Required for Structural and Functional Integrity of the Spindle Apparatus

Nathalie Colombié ^*, Christel Vérollet ^*, Paula Sampaio , André Moisand , Claudio Sunkel  , Henri-Marc Bourbon ^||, Michel Wright ^*, and Brigitte Raynaud-Messina ^*

^* Centre de Recherche en Pharmacologie-Santé, Unité Mixte de Recherche 2587, Centre National de la Recherche Scientifique-Pierre Fabre, Institut de Sciences et Technologies du Médicament de Toulouse, 31400 Toulouse, France; ^|| Centre de Biologie du Développement, Unité Mixte de Recherche 5547, Centre National de la Recherche Scientifique-Université Paul Sabatier, Institut d'Exploration Fonctionnelle des Génomes, 31062 Toulouse, France; Instituto de Biologia Molecular e Celular, Universitade do Porto, 4150-180 Porto, Portugal; Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, 4099-003 Porto, Portugal; and Institut de Pharmacologie et de Biologie Structurale, Unité Mixte de Recherche 5089, 31077 Toulouse, France

Submitted August 4, 2005; Accepted October 5, 2005
Monitoring Editor: Tim Stearns

-Tubulin, a protein critical for microtubule assembly, functions within multiprotein complexes. However, little is known about the respective role of -tubulin partners in metazoans. For the first time in a multicellular organism, we have investigated the function of Dgrip84, the Drosophila orthologue of the Saccharomyces cerevisiae -tubulin-associated protein Spc97p. Mutant analysis shows that Dgrip84 is essential for viability. Its depletion promotes a moderate increase in the mitotic index, correlated with the appearance of monopolar or unpolarized spindles, impairment of centrosome maturation, and increase of polyploid nuclei. This in vivo study is strengthened by an RNA interference approach in cultured S2 cells. Electron microscopy analysis suggests that monopolar spindles might result from a failure of centrosome separation and an unusual microtubule assembly pathway via centriolar triplets. Moreover, we point to an involvement of Dgrip84 in the spindle checkpoint regulation and in the maintenance of interphase microtubule dynamics. Dgrip84 also seems essential for male meiosis, ensuring spindle bipolarity and correct completion of cytokinesis. These data sustain that Dgrip84 is required in some aspects of microtubule dynamics and organization both in interphase and mitosis. The nature of a minimal -tubulin complex necessary for proper microtubule organization in the metazoans is discussed.

The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Address correspondence to: Brigitte Raynaud-Messina (brigitte. raynaud-messina{at}istmt.cnrs.fr).

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