A large-scale in vivo RNAi screen to identify genes involved in Notch-mediated follicle cell differentiation and cell cycle switches.
A large-scale in vivo RNAi screen to identify genes involved in Notch-mediated follicle cell differentiation and cell cycle switches.
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DOI：
10.1038/srep12328
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发表时间：
2015-07-24
影响因子：
4.6
通讯作者：
Deng WM
中科院分区：
文献类型：
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作者：
Jia D;Soylemez M;Calvin G;Bornmann R;Bryant J;Hanna C;Huang YC;Deng WM
在果蝇的卵发生过程中，卵泡细胞依次经历三个不同的细胞周期程序：有丝分裂周期、内周期和基因扩增。Notch信号在调节卵泡细胞分化和细胞周期开关中起核心作用；它的激活对于有丝分裂周期/内循环（M/E）开关至关重要。Cut是Notch信号传导和细胞周期调节因子之间的连接物，在细胞内周期阶段被Notch特异性下调。为了确定在M/E转换过程中信号通路是如何协调的，并鉴定参与卵泡细胞分化的新基因，我们通过诱导敲低基因表达和检查卵泡细胞中的Cut表达进行了体内RNAi筛选。我们筛选了2205个RNAi细胞系，发现了33个在M/E转换过程中调控Cut表达的基因。这些基因通过另外两个Notch信号下游因子（后见之明和Broad）的染色得到证实，并通过多个独立的RNAi细胞系进行了验证。我们应用基因本体软件寻找丰富的生物学意义，并将我们的结果与其他出版物进行比较，以寻找跨组织的保守基因。具体来说，我们发现前卵泡细胞比后卵泡细胞更早进入内循环，确定胰岛素- pi3k通路参与精确的M/E开关，并提出Nejire是卵子发生过程中Notch信号传导的辅助因子。
During Drosophila oogenesis, follicle cells sequentially undergo three distinct cell-cycle programs: the mitotic cycle, endocycle, and gene amplification. Notch signaling plays a central role in regulating follicle-cell differentiation and cell-cycle switches; its activation is essential for the mitotic cycle/endocycle (M/E) switch. Cut, a linker between Notch signaling and cell-cycle regulators, is specifically downregulated by Notch during the endocycle stage. To determine how signaling pathways coordinate during the M/E switch and to identify novel genes involved in follicle cell differentiation, we performed an in vivo RNAi screen through induced knockdown of gene expression and examination of Cut expression in follicle cells. We screened 2205 RNAi lines and found 33 genes regulating Cut expression during the M/E switch. These genes were confirmed with the staining of two other Notch signaling downstream factors, Hindsight and Broad, and validated with multiple independent RNAi lines. We applied gene ontology software to find enriched biological meaning and compared our results with other publications to find conserved genes across tissues. Specifically, we found earlier endocycle entry in anterior follicle cells than those in the posterior, identified that the insulin-PI3K pathway participates in the precise M/E switch, and suggested Nejire as a cofactor of Notch signaling during oogenesis.