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  • Original Article
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hSMG-1 and ATM sequentially and independently regulate the G1 checkpoint during oxidative stress

Oncogene volume 27, pages 4065–4074 (2008)Cite this article

Abstract

Genotoxic stress activates the phosphatidylinositol 3-kinase-like kinases (PIKKs) that phosphorylate proteins involved in cell cycle arrest, DNA repair and apoptosis. Previous work showed that the PIKK ataxia telangiectasia mutated (ATM) but not ATM and Rad3 related phosphorylates p53 (Ser15) during hyperoxia, a model of prolonged oxidative stress and DNA damage. Here, we show hSMG-1 is responsible for the rapid and early phosphorylation of p53 (Ser15) and that ATM helps maintain phosphorylation after 24 h. Despite reduced p53 phosphorylation and abundance in cells depleted of hSMG-1 or ATM, levels of the p53 target p21 were still elevated and the G1 checkpoint remained intact. Conditional overexpression of p21 in p53-deficient cells revealed that hyperoxia also stimulates wortmannin-sensitive degradation of p21. siRNA depletion of hSMG-1 or ATM restored p21 stability and the G1 checkpoint during hyperoxia. These findings establish hSMG-1 as a proximal regulator of DNA damage signaling and reveal that the G1 checkpoint is tightly regulated during prolonged oxidative stress by both PIKK-dependent synthesis and proteolysis of p21.

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Acknowledgements

We thank Peter Vitiello, Melissa Wu, and Jennifer Gewandter for critiquing this manuscript. This work was funded in part by National Institutes of Health Grants HL-67392 (MA O’Reilly) and ES-01247. NIH training grant ES-07026 supported S Gehen.

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Authors and Affiliations

  1. Department of Environmental Medicine, The University of Rochester, Rochester, NY, USA

    S C Gehen

  2. Department of Pediatrics, School of Medicine and Dentistry, The University of Rochester, Rochester, NY, USA

    R J Staversky & M A O'Reilly

  3. Department of Biochemistry and Biophysics, The University of Rochester, Rochester, NY, USA

    R A Bambara

  4. Department of Radiation Oncology, The University of Rochester, Rochester, NY, USA

    P C Keng

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  1. S C Gehen
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  2. R J Staversky
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  3. R A Bambara
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  4. P C Keng
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  5. M A O'Reilly
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Correspondence to M A O'Reilly.

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Gehen, S., Staversky, R., Bambara, R. et al. hSMG-1 and ATM sequentially and independently regulate the G1 checkpoint during oxidative stress. Oncogene 27, 4065–4074 (2008). https://doi.org/10.1038/onc.2008.48

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