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. 1999 Feb 1;18(3):763–770. doi: 10.1093/emboj/18.3.763

p53 DNA binding can be modulated by factors that alter the conformational equilibrium.

K G McLure 1, P W Lee 1
PMCID: PMC1171169  PMID: 9927436

Abstract

The p53 tumor suppressor protein is a dimer of dimers that binds its consensus DNA sequence (containing two half-sites) as a pair of clamps. We show here that after one wild-type dimer of a tetramer binds to a half-site on the DNA, the other (unbound) dimer can be in either the wild-type or the mutant conformation. An equilibrium state between these two conformations exists and can be modulated by two types of regulators. One type modifies p53 biochemically and determines the intrinsic balance of the equilibrium. The other type of regulator binds directly to one or both dimers in a p53 tetramer, trapping each dimer in one or the other conformation. In the wild-type conformation, the second dimer can bind to the second DNA half-site, resulting in drastically enhanced stability of the p53-DNA complex. Importantly, a genotypically mutant p53 can also be in equilibrium with the wild-type conformation, and when trapped in this conformation can bind DNA.

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Selected References

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  1. Blagosklonny M. V., Toretsky J., Bohen S., Neckers L. Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90. Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8379–8383. doi: 10.1073/pnas.93.16.8379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chen Y., Chen P. L., Lee W. H. Hot-spot p53 mutants interact specifically with two cellular proteins during progression of the cell cycle. Mol Cell Biol. 1994 Oct;14(10):6764–6772. doi: 10.1128/mcb.14.10.6764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cho Y., Gorina S., Jeffrey P. D., Pavletich N. P. Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. Science. 1994 Jul 15;265(5170):346–355. doi: 10.1126/science.8023157. [DOI] [PubMed] [Google Scholar]
  4. Cook A., Milner J. Evidence for allosteric variants of wild-type p53, a tumour suppressor protein. Br J Cancer. 1990 Apr;61(4):548–552. doi: 10.1038/bjc.1990.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Friend S. p53: a glimpse at the puppet behind the shadow play. Science. 1994 Jul 15;265(5170):334–335. doi: 10.1126/science.8023155. [DOI] [PubMed] [Google Scholar]
  6. Gannon J. V., Greaves R., Iggo R., Lane D. P. Activating mutations in p53 produce a common conformational effect. A monoclonal antibody specific for the mutant form. EMBO J. 1990 May;9(5):1595–1602. doi: 10.1002/j.1460-2075.1990.tb08279.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gorina S., Pavletich N. P. Structure of the p53 tumor suppressor bound to the ankyrin and SH3 domains of 53BP2. Science. 1996 Nov 8;274(5289):1001–1005. doi: 10.1126/science.274.5289.1001. [DOI] [PubMed] [Google Scholar]
  8. Hainaut P., Milner J. Interaction of heat-shock protein 70 with p53 translated in vitro: evidence for interaction with dimeric p53 and for a role in the regulation of p53 conformation. EMBO J. 1992 Oct;11(10):3513–3520. doi: 10.1002/j.1460-2075.1992.tb05434.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hainaut P., Milner J. Redox modulation of p53 conformation and sequence-specific DNA binding in vitro. Cancer Res. 1993 Oct 1;53(19):4469–4473. [PubMed] [Google Scholar]
  10. Hainaut P., Rolley N., Davies M., Milner J. Modulation by copper of p53 conformation and sequence-specific DNA binding: role for Cu(II)/Cu(I) redox mechanism. Oncogene. 1995 Jan 5;10(1):27–32. [PubMed] [Google Scholar]
  11. Halazonetis T. D., Davis L. J., Kandil A. N. Wild-type p53 adopts a 'mutant'-like conformation when bound to DNA. EMBO J. 1993 Mar;12(3):1021–1028. doi: 10.1002/j.1460-2075.1993.tb05743.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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. 1993 Dec 15;12(13):5057–5064. doi: 10.1002/j.1460-2075.1993.tb06199.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hall A. R., Milner J. Structural and kinetic analysis of p53-DNA complexes and comparison of human and murine p53. Oncogene. 1995 Feb 2;10(3):561–567. [PubMed] [Google Scholar]
  14. Hollstein M., Shomer B., Greenblatt M., Soussi T., Hovig E., Montesano R., Harris C. C. Somatic point mutations in the p53 gene of human tumors and cell lines: updated compilation. Nucleic Acids Res. 1996 Jan 1;24(1):141–146. doi: 10.1093/nar/24.1.141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hupp T. R., Meek D. W., Midgley C. A., Lane D. P. Regulation of the specific DNA binding function of p53. Cell. 1992 Nov 27;71(5):875–886. doi: 10.1016/0092-8674(92)90562-q. [DOI] [PubMed] [Google Scholar]
  16. Hupp T. R., Sparks A., Lane D. P. Small peptides activate the latent sequence-specific DNA binding function of p53. Cell. 1995 Oct 20;83(2):237–245. doi: 10.1016/0092-8674(95)90165-5. [DOI] [PubMed] [Google Scholar]
  17. Iwabuchi K., Bartel P. L., Li B., Marraccino R., Fields S. Two cellular proteins that bind to wild-type but not mutant p53. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6098–6102. doi: 10.1073/pnas.91.13.6098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jayaraman L., Murthy K. G., Zhu C., Curran T., Xanthoudakis S., Prives C. Identification of redox/repair protein Ref-1 as a potent activator of p53. Genes Dev. 1997 Mar 1;11(5):558–570. doi: 10.1101/gad.11.5.558. [DOI] [PubMed] [Google Scholar]
  19. Koshland D. E., Jr, Némethy G., Filmer D. Comparison of experimental binding data and theoretical models in proteins containing subunits. Biochemistry. 1966 Jan;5(1):365–385. doi: 10.1021/bi00865a047. [DOI] [PubMed] [Google Scholar]
  20. Legros Y., Meyer A., Ory K., Soussi T. Mutations in p53 produce a common conformational effect that can be detected with a panel of monoclonal antibodies directed toward the central part of the p53 protein. Oncogene. 1994 Dec;9(12):3689–3694. [PubMed] [Google Scholar]
  21. MONOD J., WYMAN J., CHANGEUX J. P. ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL. J Mol Biol. 1965 May;12:88–118. doi: 10.1016/s0022-2836(65)80285-6. [DOI] [PubMed] [Google Scholar]
  22. McLure K. G., Lee P. W. A PAb240+ conformation of wild type p53 binds DNA. Oncogene. 1996 Sep 19;13(6):1297–1303. [PubMed] [Google Scholar]
  23. McLure K. G., Lee P. W. How p53 binds DNA as a tetramer. EMBO J. 1998 Jun 15;17(12):3342–3350. doi: 10.1093/emboj/17.12.3342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Milner J., Cook A., Sheldon M. A new anti-p53 monoclonal antibody, previously reported to be directed against the large T antigen of simian virus 40. Oncogene. 1987;1(4):453–455. [PubMed] [Google Scholar]
  25. Milner J., Cook A. The cellular tumour antigen p53: evidence for transformation-related, immunological variants of p53. Virology. 1986 Oct 15;154(1):21–30. doi: 10.1016/0042-6822(86)90426-5. [DOI] [PubMed] [Google Scholar]
  26. Milner J. Flexibility: the key to p53 function? Trends Biochem Sci. 1995 Feb;20(2):49–51. doi: 10.1016/s0968-0004(00)88954-9. [DOI] [PubMed] [Google Scholar]
  27. Milner J., Medcalf E. A. Temperature-dependent switching between "wild-type" and "mutant" forms of p53-Val135. J Mol Biol. 1990 Dec 5;216(3):481–484. doi: 10.1016/0022-2836(90)90371-R. [DOI] [PubMed] [Google Scholar]
  28. Milner J., Watson J. V. Addition of fresh medium induces cell cycle and conformation changes in p53, a tumour suppressor protein. Oncogene. 1990 Nov;5(11):1683–1690. [PubMed] [Google Scholar]
  29. Rolley N., Butcher S., Milner J. Specific DNA binding by different classes of human p53 mutants. Oncogene. 1995 Aug 17;11(4):763–770. [PubMed] [Google Scholar]
  30. Vojtesek B., Dolezalova H., Lauerova L., Svitakova M., Havlis P., Kovarik J., Midgley C. A., Lane D. P. Conformational changes in p53 analysed using new antibodies to the core DNA binding domain of the protein. Oncogene. 1995 Jan 19;10(2):389–393. [PubMed] [Google Scholar]
  31. Waterman J. L., Shenk J. L., Halazonetis T. D. The dihedral symmetry of the p53 tetramerization domain mandates a conformational switch upon DNA binding. EMBO J. 1995 Feb 1;14(3):512–519. doi: 10.1002/j.1460-2075.1995.tb07027.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wolkowicz R., Elkind N. B., Ronen D., Rotter V. The DNA binding activity of wild type p53 is modulated by blocking its various antigenic epitopes. Oncogene. 1995 Mar 16;10(6):1167–1174. [PubMed] [Google Scholar]
  33. Zhang W., Funk W. D., Wright W. E., Shay J. W., Deisseroth A. B. Novel DNA binding of p53 mutants and their role in transcriptional activation. Oncogene. 1993 Sep;8(9):2555–2559. [PubMed] [Google Scholar]

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