Dobbelaere, J and Josue, F and Suijkerbuijk, S and Baum, B and Tapon, N and 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.
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.
|Title:||A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila|
|Open access status:||An open access publication|
|Keywords:||PROTEIN PHOSPHATASE 2A, MITOTIC SPINDLE POLES, AURORA-A KINASE, C-ELEGANS, GAMMA-TUBULIN, CELL-CYCLE, PERICENTRIOLAR MATERIAL, MICROTUBULE NUCLEATION, CAENORHABDITIS-ELEGANS, SPD-2|
|UCL classification:||UCL > School of Life and Medical Sciences > Faculty of Life Sciences > Laboratory for Molecular Cell Biology|
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