Abstract
Defective S-phase checkpoint activation results in an inability to downregulate DNA replication following genotoxic insult such as exposure to ionizing radiation. This 'radioresistant DNA synthesis' (RDS) is a phenotypic hallmark of ataxia–telangiectasia1, a cancer-prone disorder caused by mutations in ATM2. The mismatch repair system principally corrects nucleotide mismatches that arise during replication3. Here we show that the mismatch repair system is required for activation of the S-phase checkpoint in response to ionizing radiation. Cells deficient in mismatch repair proteins showed RDS, and restoration of mismatch repair function restored normal S-phase checkpoint function. Catalytic activation of ATM and ATM-mediated phosphorylation of the protein NBS1 (also called nibrin) occurred independently of mismatch repair. However, ATM-dependent phosphorylation and activation of the checkpoint kinase CHK2 and subsequent degradation of its downstream target, CDC25A, was abrogated in cells lacking mismatch repair. In vitro and in vivo approaches both show that MSH2 binds to CHK2 and that MLH1 associates with ATM. These findings indicate that the mismatch repair complex formed at the sites of DNA damage facilitates the phosphorylation of CHK2 by ATM, and that defects in this mechanism form the molecular basis for the RDS observed in cells deficient in mismatch repair.
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Acknowledgements
The authors thank S. Elledge for supplying the GST–CHK2 plasmid, J. Lazo for the GST–CDC25A plasmid and B. Gebhardt for access to the 137Cs irradiator. This work was supported by grants from the American Cancer Society to K.D.B. and R.B. and the US National Institutes of Health to R.B.
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Brown, K., Rathi, A., Kamath, R. et al. The mismatch repair system is required for S-phase checkpoint activation. Nat Genet 33, 80–84 (2003). https://doi.org/10.1038/ng1052
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DOI: https://doi.org/10.1038/ng1052
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