Identification of pathways regulating cell size and cell-cycle progression by RNAi
Identification of pathways regulating cell size and cell-cycle progression by RNAi
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DOI：
10.1038/nature04469
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发表时间：
2006-02-23
期刊：
影响因子：
64.8
通讯作者：
Taipale, J
中科院分区：
文献类型：
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作者：
Björklund, M;Taipale, M;Taipale, J
真核细胞周期的许多高通量功能丧失分析依赖于单细胞酵母菌种酿酒酵母和粟酒裂殖酵母。然而，在多细胞生物体中，额外的控制机制调节细胞周期以指定生物体及其组成器官的大小（1）。为了鉴定这些基因，我们用流式细胞术分析了70%果蝇基因（包括90%人类保守基因）功能丧失对S2细胞细胞周期进程的影响。为了解决冗余问题，我们还同时靶向了参与蛋白质磷酸化的基因及其同源物。我们确定了控制细胞大小，胞质分裂，细胞死亡和/或凋亡，以及细胞周期的G1和G2/M期的基因。基于这些表型通过无监督的层次聚类将基因分类为通路表明，除了经典的调节机制如Myc/Max、Cyclin/Cdk和E2 F之外，S2细胞中的细胞周期进程还受囊泡和核转运蛋白、COP 9信号体活性和四种细胞外信号调节通路（Wnt、p38 β MAPK、FRAP/TOR和JAK/STAT）控制。此外，通过同时分析几种表型，我们确定了一个翻译调节因子，eIF-3 p66，专门影响细胞周期蛋白/Cdk途径的活动。
Many high-throughput loss-of-function analyses of the eukaryotic cell cycle have relied on the unicellular yeast species Saccharomyces cerevisiae and Schizosaccharomyces pombe. In multicellular organisms, however, additional control mechanisms regulate the cell cycle to specify the size of the organism and its constituent organs(1). To identify such genes, here we analysed the effect of the loss of function of 70% of Drosophila genes (including 90% of genes conserved in human) on cell-cycle progression of S2 cells using flow cytometry. To address redundancy, we also targeted genes involved in protein phosphorylation simultaneously with their homologues. We identify genes that control cell size, cytokinesis, cell death and/or apoptosis, and the G1 and G2/M phases of the cell cycle. Classification of the genes into pathways by unsupervised hierarchical clustering on the basis of these phenotypes shows that, in addition to classical regulatory mechanisms such asMyc/Max, Cyclin/Cdk and E2F, cell-cycle progression in S2 cells is controlled by vesicular and nuclear transport proteins, COP9 signalosome activity and four extracellular-signal-regulated pathways (Wnt, p38 beta MAPK, FRAP/TOR and JAK/STAT). In addition, by simultaneously analysing several phenotypes, we identify a translational regulator, eIF-3p66, that specifically affects the Cyclin/Cdk pathway activity.