Functional genomic screens identify CINP as a genome maintenance protein
Functional genomic screens identify CINP as a genome maintenance protein
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DOI：
10.1073/pnas.0909345106
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发表时间：
2009-11-17
影响因子：
11.1
通讯作者：
Cortez, David
中科院分区：
文献类型：
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作者：
Lovejoy, Courtney A.;Xu, Xin;Cortez, David
DNA 损伤反应 (DDR) 在维持基因组完整性方面发挥着关键作用，并通过促进细胞周期停滞、DNA 修复和细胞凋亡来充当肿瘤发生的屏障。 DDR 不仅会被诱导 DNA 损伤的基因毒剂激活，还会在由激活的癌基因和失活的抑癌基因引起的异常细胞分裂周期中被激活。在这里，我们使用人类细胞中的 RNAi 和 cDNA 过表达筛选来识别当解除管制时会导致 DDR 激活的基因。 RNAi 筛选鉴定出 73 个基因，当这些基因在至少两种细胞类型中沉默时，会导致 DDR 激活。沉默其中几个基因也会导致微核频率增加，微核是遗传不稳定细胞的标志。 cDNA 筛选鉴定出 97 个基因，这些基因在没有任何外源性基因毒性剂的情况下过度表达时会诱导 DDR 激活，其中与癌症相关的基因过多。二次 RNAi 筛选将 CDK2 相互作用蛋白 (CINP) 鉴定为细胞周期检查点蛋白。 CINP 与 ATR 相互作用蛋白相互作用，调节 ATR 依赖性信号传导、复制应激抵抗力和 G2 检查点完整性。
The DNA damage response (DDR) has a critical role in maintaining genome integrity and serves as a barrier to tumorigenesis by promoting cell-cycle arrest, DNA repair, and apoptosis. The DDR is activated not only by genotoxic agents that induce DNA damage, but also during aberrant cell-division cycles caused by activated oncogenes and inactivated tumor suppressors. Here we use RNAi and cDNA overexpression screens in human cells to identify genes that, when deregulated, lead to activation of the DDR. The RNAi screen identified 73 genes that, when silenced in at least two cell types, cause DDR activation. Silencing several of these genes also caused an increased frequency of micronuclei, a marker of genetically unstable cells. The cDNA screen identified 97 genes that when overexpressed induce DDR activation in the absence of any exogenous genotoxic agent, with an overrepresentation of genes linked to cancer. Secondary RNAi screens identified CDK2-interacting protein (CINP) as a cell-cycle checkpoint protein. CINP interacts with ATR-interacting protein and regulates ATR-dependent signaling, resistance to replication stress, and G2 checkpoint integrity.