Abstract
The p53 tumour-suppressor protein is a sequence-specific DNA-binding transcription factor that induces cell cycle arrest or apoptosis in response to genotoxic stress1,2,3,4,5,6. Activation of p53 by DNA-damaging agents is critical for eliminating cells with damaged genomic DNA and underlies the apoptotic response of human cancers treated with ionizing radiation (IR) and radiomimetic drugs7,8. The molecular mechanisms by which DNA damage activates p53 have not been elucidated. Both the levels of p53 protein and its affinity for specific DNA sequences increase in response to genotoxic stress6,9,10. In vitro, the affinity of p53 for DNA is regulated by its carboxy-terminus11,12,13. We therefore examined whether this region of p53 is targeted by DNA-damage signalling pathways in vivo. In non-irradiated cells, serines 376 and 378 of p53 were phosphorylated. IR led to dephosphorylation of Ser376, creating a consensus binding site for 14-3-3 proteins and leading to association of p53 with 14-3-3. In turn, this increased the affinity of p53 for sequence-specific DNA. Consistent with the lack of p53 activation by IR in ataxia telangiectasia (AT; Refs 14,15), neither Ser376 dephosphorylation, nor the interaction of p53 with 14-3-3 proteins occurred in AT cells.
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References
Levine, A.J. p53, the cellular gatekeeper for growth and division. Cell 88, 323–331 (1997)
Kastan, M.B., Onyekwere, O., Sidransky, D., Vogelstein, B. & Craig, R.W. Participation of p53 protein in the cellular response to DNA damage. Cancer Res. 51, 6304–6311 (1991)
Kuerbitz, S.J., Plunkett, B.S., Walsh, W.V. & Kastan, M.B. Wild-type p53 is a cell cycle checkpoint determinant following irradiation . Proc. Natl. Acad. Sci. USA 89, 7491–7495 (1992)
Clarke, A.R. et al. Thymocyte apoptosis induced by p53-dependent and independent pathways. Nature 362, 849–852 ( 1993)
Lowe, S.W., Schmitt, E.M., Smith, S.W., Osborne, B.A. & Jacks, T. p53 is required for radiation-induced apoptosis in mouse thymocytes. Nature 362, 847–849 (1993)
Maltzman, W. & Czyzyk, L. UV irradiation stimulates levels of p53 cellular tumour antigen in nontransformed mouse cells. Mol. Cell. Biol. 4, 1689–1694 (1984)
Lowe, S.W. et al. p53 status and the efficacy of cancer therapy in vivo. Science 266, 807–810 ( 1994)
Fisher, D.E. Apoptosis in cancer therapy: crossing the threshold. Cell 78, 539–542 (1994)
Chernov, M. & Stark, G.R. The p53 activation and apoptosis induced by DNA damage are reversibly inhibited by salicylate. Oncogene 14, 2503–2510 ( 1997)
Haapajarvi, T., Pitkanen, K., Tsubari, M. & Laiho, M. p53 transactivation and protein accumulation are independently regulated by UV light in different phases of the cell cycle. Mol. Cell. Biol. 17, 3074–3080 (1997)
Hupp, T.R., Meek, D.W., Midgley, C.A. & Lane D.P. Regulation of the specific DNA binding function of p53. Cell 71, 875–886 (1992)
Halazonetis, T.D., Davis, L.J. & Kandil, A.N. Wild-type p53 adopts a "mutant"-like conformation when bound to DNA. EMBO J. 12, 1021–1028 (1993)
Waterman, J.L.F., Shenk, J.L. & Halazonetis, T.D. The dihedral symmetry of the p53 tetramerization domain mandates a conformational switch upon DNA binding. EMBO J. 14, 512–519 (1995)
Kastan, M.B. et al. A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 71, 587–597 (1992)
Khanna, K.K. & Lavin, M.F. Ionizing radiation and UV induction of p53 protein by different pathways in ataxia-telangiectasia cells. Oncogene 8, 3307–3312 ( 1993)
Liu, Z.G. et al. Three distinct signalling responses by murine fibroblasts to genotoxic stress . Nature 384, 273–276 (1996)
Savitsky, K. et al. A single Ataxia Telangiectasia gene with a product similar to PI-3 kinase . Science 268, 1749–1753 (1995)
Shieh, S.Y., Ikeda, M., Taya, Y. & Prives, C. DNA damage-induced phosphorylation of p53 alleviates inhibition by Mdm2. Cell 91, 325–334 (1997)
Siliciano, J.D. et al. DNA damage induces phosphorylation of the amino terminus of p53. Genes Dev. 11, 3471–3481 (1997)
Haupt, Y., Maya, R., Kazaz, A. & Oren, M. Mdm2 promotes the rapid degradation of p53. Nature 387, 296–299 (1997)
Kubbutat, M.H.G., Jones, S.N. & Vousden, K.H. Regulation of p53 stability by mdm2. Nature 387, 299–303 ( 1997)
Delphin, C. & Baudier, J. The protein kinase C activator, phorbol ester, cooperates with the wild-type p53 species of ras-transformed embryo fibroblasts growth arrest. J. Biol. Chem. 269, 29579–29587 (1994)
Soussi, T. & May, P. Structural aspects of the p53 protein in relation to gene evolution: a second look. J. Mol. Biol. 260, 623–637 (1996)
Aitken, A. 14-3-3 and its possible role in co-ordinating multiple signalling pathways. Trends Cell Biol. 6, 341–347 ( 1996)
Yaffe, M.B. et al. The structural basis for 14-3-3: phosphopeptide binding specificity. Cell 91, 961–971 ( 1997)
Muslin, A.J., Tanner, J.W., Allen, P.M. & Shaw, A.S. Interaction of 14-3-3 with signaling proteins is mediated by the recognition of phosphoserine. Cell 84, 889–897 ( 1996)
Hermeking, H. et al. 14-3-3σ is a p53-regulated inhibitor of G2/M progression. Mol. Cell 1, 3–11 ( 1997)
Ford, J.C. et al. 14-3-3 protein homologs required for the DNA damage checkpoint in fission yeast. Science 265, 533–535 (1994)
Peng, C.Y. et al. Mitotic and G2 checkpoint control: regulation of 14-3-3 protein binding by phosphorylation of cdc25c on serine-216. Science 277 , 1501–1505 (1997)
Waterman, M.J.F., Waterman, J.L.F. & Halazonetis, T.D. An engineered four-stranded coiled coil substitutes for the tetramerization domain of wild-type p53 and alleviates transdominant inhibition by tumour-derived p53 mutants. Cancer Res. 56, 158–163 (1996)
Acknowledgements
We thank G. Rovera, C. Prives, P. Leder, J. Shafer, F. Rauscher III, D. George, T. Roberts, L. Chodosh, R. Muschel, N. Chehab, D. Scolnick and M. Lien for helpful discussions and reagents. Financial support was provided by the American Cancer Society and the W. W. Smith Charitable Trust (T.D.H.), an NIH Program Project for Breast Cancer Pilot Studies to the University of Pennsylvania, the Wistar Institute NIH training grant (M.J.F.W.) and the National Cancer Center (E.S.S.).
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Waterman, M., Stavridi, E., Waterman, J. et al. ATM-dependent activation of p53 involves dephosphorylation and association with 14-3-3 proteins. Nat Genet 19, 175–178 (1998). https://doi.org/10.1038/542
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DOI: https://doi.org/10.1038/542
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