ATR inhibition rewires cellular signaling networks induced by replication stress
ATR inhibition rewires cellular signaling networks induced by replication stress
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DOI：
10.1002/pmic.201500172
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发表时间：
2016-02-01
期刊：
影响因子：
3.4
通讯作者：
Beli, Petra
中科院分区：
文献类型：
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作者：
Wagner, Sebastian A.;Oehler, Hannah;Beli, Petra
复制叉的减慢或停滞通常被称为复制应激，并且由多种原因引起，例如DNA损伤、核苷酸耗尽、RNA-DNA杂交和癌基因激活。共济失调毛细血管扩张和Rad 3相关激酶（ATR）在细胞对复制应激的反应中起重要作用，并且ATR的抑制已经成为用于治疗表现出高水平复制应激的癌症的治疗策略。然而，复制应激诱导的细胞信号转导和ATR的底物谱尚未得到系统的研究。在这项研究中，我们采用定量MS为基础的蛋白质组学来定义后核苷酸耗尽诱导的复制应激和复制叉崩溃ATR抑制细胞信号。我们证明，复制压力的结果增加磷酸化的一个子集的蛋白质，其中许多参与RNA剪接和转录，以前没有与细胞复制应激反应。此外，我们的数据揭示了复制应激后的ATR依赖性磷酸化，并在MCM 6、TOPBP 1、RAD 51 AP 1和PSMD 4上发现了新的推定ATR靶位点。我们确定ATR抑制重新布线由复制应激诱导的细胞信号网络，并导致ATM驱动的双链断裂修复信号的激活。
The slowing down or stalling of replication forks is commonly known as replication stress and arises from multiple causes such as DNA lesions, nucleotide depletion, RNA-DNA hybrids, and oncogene activation. The ataxia telangiectasia and Rad3-related kinase (ATR) plays an essential role in the cellular response to replication stress and inhibition of ATR has emerged as therapeutic strategy for the treatment of cancers that exhibit high levels of replication stress. However, the cellular signaling induced by replication stress and the substrate spectrum of ATR has not been systematically investigated. In this study, we employed quantitative MS-based proteomics to define the cellular signaling after nucleotide depletion-induced replication stress and replication fork collapse following ATR inhibition. We demonstrate that replication stress results in increased phosphorylation of a subset of proteins, many of which are involved in RNA splicing and transcription and have previously not been associated with the cellular replication stress response. Furthermore, our data reveal the ATR-dependent phosphorylation following replication stress and discover novel putative ATR target sites on MCM6, TOPBP1, RAD51AP1, and PSMD4. We establish that ATR inhibition rewires cellular signaling networks induced by replication stress and leads to the activation of the ATM-driven double-strand break repair signaling.