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  • Original Article
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Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint

Oncogene volume 25, pages 5921–5932 (2006)Cite this article

Abstract

The S-phase DNA damage checkpoint is activated by DNA damage to delay DNA synthesis allowing time to resolve the replication block. We previously discovered the p53-dependent S-phase DNA damage checkpoint in mouse zygotes fertilized with irradiated sperm. Here, we report that the same p53 dependency holds in mouse embryonic fibroblasts (MEFs) at low doses of irradiation. DNA synthesis in p53 wild-type (WT) MEFs was suppressed in a biphasic manner in which a sharp decrease below 2.5 Gy was followed by a more moderate decrease up to 10 Gy. In contrast, p53−/− MEFs exhibited radioresistant DNA synthesis below 2.5 Gy whereas the cells retained the moderate suppression above 5 Gy. DNA fiber analysis revealed that 1 Gy irradiation suppressed replication fork progression in p53 WT MEFs, but not in p53−/− MEFs. Proliferating cell nuclear antigen (PCNA), clamp loader of DNA polymerase, was phosphorylated in WT MEFs after 1 Gy irradiation and redistributed to form foci in the nuclei. In contrast, PCNA was not phosphorylated and dissociated from chromatin in 1 Gy-irradiated p53−/− MEFs. These results demonstrate that the novel low-dose-specific p53-dependent S-phase DNA damage checkpoint is likely to regulate the replication fork movement through phosphorylation of PCNA.

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Acknowledgements

We thank Drs Donald MacPhee, Martin Levin, Yosef Shiloh and Mirit I Aladjem for critical reading of the manuscript. This work was supported by a Grant-in-aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan and by a grant from Nuclear Safety Research Association.

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Authors and Affiliations

  1. Department of Late Effect Studies, Radiation Biology Center, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto, Japan

    T Shimura, M Toyoshima, S K Adiga, H Nagai & O Niwa

  2. Department of Radiation System Biology, Radiation Biology Center, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto, Japan

    T Kunoh

  3. Graduate School of Biosphere Science, Hiroshima University, Hiroshima, Japan

    N Shimizu

  4. Medical Research Institute, Kanazawa Medical University, Uchinada-machi, Kahoku-gun, Ishikawa, Japan

    M Inoue

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  1. T Shimura
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Correspondence to T Shimura.

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Shimura, T., Toyoshima, M., Adiga, S. et al. Suppression of replication fork progression in low-dose-specific p53-dependent S-phase DNA damage checkpoint. Oncogene 25, 5921–5932 (2006). https://doi.org/10.1038/sj.onc.1209624

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