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DNA-dependent protein kinase acts upstream of p53 in response to DNA damage

Nature volume 394, pages 700–704 (1998)Cite this article

Abstract

The tumour suppressor p53 becomes activated as a transcription factor in response to DNA damage1,2,3, but the mechanism for this activation is unclear. A good candidate for an upstream activator of p53 is the DNA-dependent protein kinase (DNA-PK) that depends on the presence of DNA breaks for its activity4,5,6. Here we investigate the link between DNA damage and the activation of DNA-PK and of p53. To determine whether DNA-PK is an upstream mediator of the p53 DNA-damage response, we analysed a severe combined-immunodeficiency (SCID) mouse cell line, SCGR11 (refs 7, 8), and the human glioma cell line M059J (ref. 9). Both cell lines lack any detectable DNA-PK activity. We find that p53 is incapable of binding to DNA in the absence of DNA-PK, that DNA-PK is necessary but not sufficient for activation of p53 sequence-specific DNA binding, and that this activation occurs in response to DNA damage. Our results establish DNA-PK as a link between DNA damage and p53 activation, and reveal the existence of a mammalian DNA-damage-response pathway.

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Figure 1: The effect of DNA damage on p53 DNA-binding activity in nuclear extracts from DNA-PK-deficient SCGR11 cells.
Figure 2: The effect of DNA damage on p53-dependent transactivation in the SCGR11 cells.
Figure 3: Comparing primary SCID cells and the SCRG11 SCID cell line in terms of DNA-PK activity and p53 DNA-binding activity.
Figure 4: The effect of M059K and M059J nuclear extracts on DNA-binding activity of in vitro -synthesized p53 and SCGR11-cell p53.
Figure 5: Defining the role of DNA-PK in p53 DNA-binding activity.

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Acknowledgements

We thank P. Jeggo for SCGR11 SCID cells, J. Allalunis-Turner for M059J and M059K cells, J. Milner for the wild-type murine p53 plasmid pSP6p53-Ala, B. Vogelstein for pG13CAT, J. Martinez for the pAb421 antibody, D. Chan for purifying DNA-PK, and E. Rattner and B. Carson for help with the preparation of mouse embryo fibroblasts. This work was supported by a grant (to P.W.K.L.) from the Alberta Cancer Board; K.G.M. was a recipient of the Alberta Heritage Foundation for Medical Research studentship.

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Authors and Affiliations

  1. Departments of Microbiology and Infectious Diseases, Calgary, T2N 4N1, Alberta, Canada

    Richard A. Woo, Kevin G. McLure & Patrick W. K. Lee

  2. Biological Sciences, Calgary, T2N 4N1, Alberta, Canada

    Susan P. Lees-Miller

  3. Biochemistry and Molecular Biology, University of Calgary, Calgary, T2N 4N1, Alberta, Canada

    Derrick E. Rancourt

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  1. Richard A. Woo
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Correspondence to Patrick W. K. Lee.

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Woo, R., McLure, K., Lees-Miller, S. et al. DNA-dependent protein kinase acts upstream of p53 in response to DNA damage. Nature 394, 700–704 (1998). https://doi.org/10.1038/29343

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