a genome-wide rnai screen to dissect centriole duplication and centrosome maturation in drosophila全基因组rnai屏幕在果蝇解剖中心体复制和中心体成熟.pdfVIP

PLoS BIOLOGY A Genome-Wide RNAi Screen to Dissect Centriole Duplication and Centrosome Maturation in Drosophila 1* ´2 1¤ 3 2 1* Jeroen Dobbelaere , Filipe Josue , Saskia Suijkerbuijk , Buzz Baum , Nicolas Tapon , Jordan Raff 1 The Gurdon Institute, Cambridge, United Kingdom, 2 Apoptosis and Proliferation Control Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom, 3 Research Institute Medical Research Council (MRC) Laboratory of Molecular Cell Biology, University College London (UCL), London, United Kingdom 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.