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Abstract：
The response to DNA replication stress in eukaryotes is under the control of the ataxia-telangiectasia and Rad3-related (ATR) kinase. ATR responds to single-stranded (ss) DNA to stabilize distressed DNA replication forks, modulate DNA replication firing and prevent cells with damaged DNA or incomplete DNA replication from entering into mitosis. Furthermore, inhibitors of ATR are currently in clinical development either as monotherapies or in combination with agents that perturb DNA replication. To gain a genetic view of the cellular pathways requiring ATR kinase function, we mapped genes whose mutation causes hypersensitivity to ATR inhibitors with genome-scale CRISPR/Cas9 screens. We delineate a consensus set of 117 genes enriched in DNA replication, DNA repair and cell cycle regulators that promote survival when ATR kinase activity is suppressed. We validate 14 genes from this set and report genes not previously described to modulate response to ATR inhibitors. In particular we found that the loss of the POLE3/POLE4 proteins, which are DNA polymerase ε accessory subunits, results in marked hypersensitivity to ATR inhibition. We anticipate that this 117-gene set will be useful for the identification of genes involved in the regulation of genome integrity and the characterization of new biological processes involving ATR, and may reveal biomarkers of ATR inhibitor response in the clinic.
摘要：
真核生物对DNA复制应激的反应受共济失调-毛细血管扩张和Rad3相关（ATR）激酶的控制。ATR响应单链（ss）DNA以稳定受损的DNA复制叉，调节DNA复制激发并防止DNA受损或DNA复制不完全的细胞进入有丝分裂。此外，ATR抑制剂目前作为单一疗法或与干扰DNA复制的药物组合在临床开发中。为了获得需要ATR激酶功能的细胞途径的遗传学观点，我们通过基因组规模的CRISPR/Cas9筛选定位了突变导致ATR抑制剂超敏反应的基因.我们描绘了一组共有的117个富含DNA复制的基因，当ATR激酶活性被抑制时促进存活的DNA修复和细胞周期调节剂。我们验证了该组中的14个基因，并报告了以前未描述的调节ATR抑制剂反应的基因。特别是我们发现POLE3/POLE4蛋白的缺失，它们是DNA聚合酶ε辅助亚基，导致对ATR抑制的明显超敏反应。我们预计，这个117基因集将有助于鉴定参与调节基因组完整性的基因，并表征涉及ATR的新生物过程。并可能在临床上揭示ATR抑制剂反应的生物标志物。
