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A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila

Dobbelaere, J; Josue, F; Suijkerbuijk, S; Baum, B; Tapon, N; Raff, J; (2008) A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila. PLOS BIOL , 6 (9) , Article e224. 10.1371/journal.pbio.0060224. Green and gold open access

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Abstract

Centrosomes comprise a pair of centrioles surrounded by an amorphous pericentriolar material (PCM). Here, we have performed a microscopy-based genome-wide RNA interference (RNAi) screen in Drosophila cells to identify proteins required for centriole duplication and mitotic PCM recruitment. We analysed 92% of the Drosophila genome ( 13,059 genes) and identified 32 genes involved in centrosome function. An extensive series of secondary screens classified these genes into four categories: ( 1) nine are required for centriole duplication, ( 2) 11 are required for centrosome maturation, ( 3) nine are required for both functions, and ( 4) three genes regulate centrosome separation. These 32 hits include several new centrosomal components, some of which have human homologs. In addition, we find that the individual depletion of only two proteins, Polo and Centrosomin (Cnn) can completely block centrosome maturation. Cnn is phosphorylated during mitosis in a Polo-dependent manner, suggesting that the Polo-dependent phosphorylation of Cnn initiates centrosome maturation in flies.

Type:Article
Title:A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila
Open access status:An open access publication. A version is also available from UCL Discovery.
DOI:10.1371/journal.pbio.0060224
Publisher version:http://dx.doi.org/10.1371/journal.pbio.0060224
Language:English
Additional information:© 2008 Dobbelaere et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. This research was supported by Fellowships from Cancer Research UK (JR and NT). During the course of this work, JD was supported by postdoctoral fellowship of the European Molecular Biology Organization (EMBO) and the Human Frontier Science Program. FJ was funded by a studentship from Fundação para a Ciência e a Tecnologia, Programa Doutoral em Biologia Experimental e Biomedicina (PDBEB) (Portugal). BB was funded by the Royal Society, the Ludwig Institute for Cancer Research (LICR), and the EMBO Young Investigator program (YIP).
Keywords:PROTEIN PHOSPHATASE 2A, MITOTIC SPINDLE POLES, AURORA-A KINASE, C-ELEGANS, GAMMA-TUBULIN, CELL-CYCLE, PERICENTRIOLAR MATERIAL, MICROTUBULE NUCLEATION, CAENORHABDITIS-ELEGANS, SPD-2

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