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DNA-dependent protein kinase is not required for the p53-dependent response to DNA damage

Nature volume 400, pages 81–83 (1999)Cite this article

Abstract

Damage to DNA in the cell activates the tumour-suppressor protein p53 (ref. 1), and failure of this activation leads to genetic instability and a predisposition to cancer. It is therefore crucial to understand the signal transduction mechanisms that connect DNA damage with p53 activation. The enzyme known as DNA-dependent protein kinase (DNA-PK) has been proposed to be an essential activator of p53 (refs 2, 3), but the evidence for its involvement in this pathway is controversial3,4. We now show that the p53 response is fully functional in primary mouse embryonic fibroblasts lacking DNA-PK: irradiation-induced DNA damage in these defective fibroblasts induces a normal response of p53 accumulation, phosphorylation of a p53 serine residue at position 15, nuclear localization and binding to DNA of p53. The upregulation of p53-target genes and cell-cycle arrest also occur normally. The DNA-PK-deficient cell line SCGR11 contains a homozygous mutation in the DNA-binding domain of p53, which may explain the defective response by p53 reported in this line3. Our results indicate that DNA-PK activity is not required for cells to mount a p53-dependent response to DNA damage.

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Figure 1: Cell-cycle arrest response of DNA-PK−/− MEFs following irradiation.
Figure 2: Accumulation and localization of p53 protein.
Figure 3: p53 binding and transactivation in DNA-PK−/− MEFs.

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Acknowledgements

We thank R. S. Haltiguanger and S. Tronick for help with Ab421 coupling. This work was supported by the MRC (A.P., P.A.J.), by a postdoctoral fellowship from the NIH (G.S.J.), by grants from the NIH and from the G. Harold and Leila Y. Mathers Charitable Foundation (G.M.W. and M.B.), in part by the Wellcome Trust, by the Human Frontiers Science Program (F.B.), and in part by the Leukemia Society of America and by a grant from the National Cancer Institute, NIH (G.E.T.). M.H. is an R.M. Phillips senior research fellow; G.E.T. is a scholar of the Leukemia Society of America.

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  1. Gretchen S. Jimenez, Fredrik Bryntesson and Michael Hubank: These authors contributed equally to this work

Authors and Affiliations

  1. Gene Expression Laboratory, The Salk Institute, La Jolla, 92037, California, USA

    Gretchen S. Jimenez, Michelle Beeche & Geoffrey M. Wahl

  2. Trafford Centre, University of Sussex, Brighton, BN1 9RY, Sussex, UK

    Fredrik Bryntesson & Guillermo E. Taccioli

  3. Department of Microbiology, Boston University School of Medicine, Boston, 02118, Massachusetts, USA

    Maria I. Torres-Arzayus, Anne Priestley & Penny A. Jeggo

  4. MRC Cell Mutation Unit, University of Sussex, Brighton, BN1 9RR, Sussex, UK

    Shin'ichi Saito, Kazuyasu Sakaguchi & Ettore Appella

  5. Laboratory of Cell Biology, National Cancer Institute, National Institute of Health, Bethesda, 20892, Maryland, USA

    Fredrik Bryntesson

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  1. Gretchen S. Jimenez
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Correspondence to Geoffrey M. Wahl.

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Jimenez, G., Bryntesson, F., Torres-Arzayus, M. et al. DNA-dependent protein kinase is not required for the p53-dependent response to DNA damage. Nature 400, 81–83 (1999). https://doi.org/10.1038/21913

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