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Activation of ATM and Chk2 kinases in relation to the amount of DNA strand breaks

Oncogene volume 23, pages 7691–7700 (2004)Cite this article

Abstract

The diverse checkpoint responses to DNA damage may reflect differential sensitivities by molecular components of the damage-signalling network to the type and amount of lesions. Here, we determined the kinetics of activation of the checkpoint kinases ATM and Chk2 (the latter substrate of ATM) in relation to the initial yield of genomic DNA single-strand (SSBs) and double-strand breaks (DSBs). We show that doses of γ-radiation (IR) as low as 0.25 Gy, which generate vast numbers of SSBs but only a few DSBs per cell (<8), promptly activate ATM kinase and induce the phosphorylation of the ATM substrates p53–Ser15, Nbs1–Ser343 and Chk2–Thr68. The full activation of Chk2 kinase, however, is triggered by treatments inflicting >19 DSBs per cell (e.g. 1 Gy), which cause Chk2 autophosphorylation on Thr387, Chk2-dependent accumulation of p21waf1 and checkpoint arrest in the S phase. Our results indicate that, in contrast to ATM, Chk2 activity is triggered by a greater number of DSBs, implying that, below a certain threshold level of lesions (<19 DSBs), DNA repair can occur through ATM, without enforcing Chk2-dependent checkpoints.

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Acknowledgements

We thank Drs B Vogelstein and F Bunz, The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, Johns Hopkins University, Baltimore, Maryland, for kindly providing the HCT116-Chk2−/− cells. This work was financially supported by the Italian Telethon Foundation grant GP0205/01, the Italian Association for Cancer Research (AIRC), the National Research Council (CNR, grant CU03.00416) and the Italian Ministry of Health (Ricerca Finalizzata). GB is recipient of a fellowship of the Italian Foundation for Cancer Research (FIRC).

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Authors and Affiliations

  1. Department of Experimental Oncology, Istituto Nazionale Tumori, Via G Venezian 1, Milan, 20133, Italy

    Giacomo Buscemi, Paola Perego, Nives Carenini & Domenico Delia

  2. Department of Biochemistry, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan

    Makoto Nakanishi

  3. Ospedale S Andrea, Via di Grotta Rossa 1035, Rome, 00189, Italy

    Luciana Chessa

  4. Departments of Oncology, Pediatric and Adolescent Medicine, Mayo Clinic and Foundation, Rochester, MN, USA

    Junjie Chen

  5. Molecular and Cellular Biology Division, The Queensland Institute of Medical Research, Brisbane, Australia

    KumKum Khanna

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  1. Giacomo Buscemi
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Correspondence to Domenico Delia.

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Buscemi, G., Perego, P., Carenini, N. et al. Activation of ATM and Chk2 kinases in relation to the amount of DNA strand breaks. Oncogene 23, 7691–7700 (2004). https://doi.org/10.1038/sj.onc.1207986

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