ATR inhibition rewires cellular signaling networks induced by replication stress.
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.
['Wagner|Sebastian A|SA|', 'Oehler|Hannah|H|', 'Voigt|Andrea|A|', 'Dalic|Denis|D|', 'Freiwald|Anja|A|', 'Serve|Hubert|H|', 'Beli|Petra|P|']
D000595:Amino Acid Sequence D000970:Antineoplastic Agents D064007:Ataxia Telangiectasia Mutated Proteins D002352:Carrier Proteins D002453:Cell Cycle D045744:Cell Line, Tumor D002470:Cell Survival D004247:DNA D004261:DNA Replication D004268:DNA-Binding Proteins D053263:Gene Regulatory Networks D006801:Humans D006918:Hydroxyurea D064151:Minichromosome Maintenance Complex Component 6 D008969:Molecular Sequence Data D009687:Nuclear Proteins D010006:Osteoblasts D010750:Phosphoproteins D010766:Phosphorylation D046988:Proteasome Endopeptidase Complex D025941:Protein Interaction Mapping D012326:RNA Splicing D016601:RNA-Binding Proteins D015398:Signal Transduction D013312:Stress, Physiological D014158:Transcription, Genetic
Cancer ATR kinase DNA replication stress Phosphoproteomics Quantitative MS Systems biology