Structure–based rescue of common tumor–derived p53 mutants (original) (raw)

Nature Medicine volume 2, pages 1143–1146 (1996)Cite this article

Abstract

The p53 tumor suppressor protein induces cell–cycle arrest or cell death in response to DNA–damaging agents, such as radiation and many of the chemotherapeutics used in cancer therapy1,2. The function of p53 is dependent on its ability to bind DNA in a sequence–specific manner3, but in one–half of all human tumors, its sequence–specific DNA binding domain is compromised by single–amino acid substitutions4. The nature of these substitutions, which target residues that directly contact DNA or that stabilize the structure of the DNA binding domain5, has raised concerns as to whether the function of p53 mutants could ever be rescued6. Nevertheless, pharmaceuticals that restore function to p53 mutants could specifically suppress proliferation of cancer cells in patients. To determine whether tumor–derived p53 mutants are irreversibly inactivated, we introduced basic residues in their DNA binding domains, aiming to establish novel contacts between p53 and the DNA phosphate backbone. In three of the seven most common p53 mutants, replacement of Thr284 with Arg significantly enhanced DNA binding affinity, without affecting DNA binding specificity, and rescued their transactivation and tumor suppressor functions. Thus, many tumor–derived p53 mutants retain their sequence–specific DNA binding determinants and can be activated to suppress tumor growth.

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References

  1. 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).
    CAS Google Scholar
  2. Lowe, S.W. et al. p53 status and the efficacy of cancer therapy in vivo. Science 266, 807–810 (1994).
    Article CAS Google Scholar
  3. Ko, L.J. & Prives, C. p53: Puzzle and paradigm. Genes Dev. 10, 1054–1072 (1996).
    Article CAS Google Scholar
  4. Hollstein, M., Sidransky, D., Vogelstein, B. & Harris, C.C. p53 mutations in human cancers. Science 253, 49–53 (1991).
    Article CAS Google Scholar
  5. Cho, Y., Gorina, S., Jeffrey, P.D. & Pavletich, N.P. Crystal structure of a p53 tumor suppressor-DNA complex: Understanding tumorigenic mutations. Science 265, 346–355 (1994).
    Article CAS Google Scholar
  6. Friend, S. p53: A glimpse at the puppet behind the shadow play. Science 265, 334–335 (1994).
    Article CAS Google Scholar
  7. 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).
    Article CAS Google Scholar
  8. 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).
    Article CAS Google Scholar
  9. 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).
    Article CAS Google Scholar
  10. Halazonetis, T.D. & Kandil, A.N. Conformational shifts propagate from the oligomerization domain of p53 to its tetrameric DNA binding domain and restore DNA binding to select p53 mutants. EMBO J. 12, 5057–5064 (1993).
    Article CAS Google Scholar
  11. Hupp, T.R., Meek, D.W., Midgley, C.A. & Lane, D.P. Activation of the cryptic DNA binding function of mutant forms of p53. Nucleic Acids Res. 21, 3167–3174 (1993).
    Article CAS Google Scholar
  12. Niewolik, D., Vojtesek, B. & Kovarik, J. p53 derived from human tumour cell lines and containing distinct point mutations can be activated to bind its consensus target sequence. Oncogene 10, 881–890 (1995).
    CAS PubMed Google Scholar
  13. Finlay, C.A. et al. Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life. Mol Cell. Biol. 8, 531–539 (1988).
    Article CAS Google Scholar
  14. El-Deiry, W.S. et al. WAF1, a potential mediator of p53 tumor suppression. Cell 75, 817–825 (1993).
    Article CAS Google Scholar
  15. 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).
    Article CAS Google Scholar
  16. Kern, S.E. et al. Oncogenic forms of p53 inhibit p53-regulated gene expression. Science 256, 827–830 (1992).
    Article CAS Google Scholar
  17. Finlay, C.A., Hinds, P.W. & Levine, A.J. The p53 proto-oncogene can act as a suppressor of transformation. Cell 57, 1083–1093 (1989).
    Article CAS Google Scholar
  18. 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 tumor-derived p53 mutants. Cancer Res. 56, 158–163 (1996).
    CAS PubMed Google Scholar
  19. Symonds, H. et al. p53-dependent apoptosis suppresses tumor growth and progression in vivo. Cell 78, 703–711 (1994).
    Article CAS Google Scholar
  20. Graeber, T.G. et al. Hypoxia-mediated selection of cells with diminished apoptotic potential in solid tumours. Nature 379, 88–91 (1996).
    Article CAS Google Scholar

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Author notes

  1. Ania M. Wieczorek and Jennifer L.F. Waterman: The first two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Oncology, the Wistar Institute, Philadelphia, Pennsylvania, 19104-4268, USA
    Ania M. Wieczorek, Jennifer L.F. Waterman, Matthew J.F. Waterman & Thanos D. Halazonetis
  2. Program in Molecular Biology and Genetics, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA
    Matthew J.F. Waterman
  3. Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, 19104, USA
    Thanos D. Halazonetis

Authors

  1. Ania M. Wieczorek
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  2. Jennifer L.F. Waterman
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  3. Matthew J.F. Waterman
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  4. Thanos D. Halazonetis
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Wieczorek, A., Waterman, J., Waterman, M. et al. Structure–based rescue of common tumor–derived p53 mutants.Nat Med 2, 1143–1146 (1996). https://doi.org/10.1038/nm1096-1143

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