Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1997 Nov;17(11):6255–6264. doi: 10.1128/mcb.17.11.6255

Reciprocal interference between the sequence-specific core and nonspecific C-terminal DNA binding domains of p53: implications for regulation.

M E Anderson 1, B Woelker 1, M Reed 1, P Wang 1, P Tegtmeyer 1
PMCID: PMC232476  PMID: 9343386

Abstract

The tumor suppressor p53 has two DNA binding domains: a central sequence-specific domain and a C-terminal sequence-independent domain. Here, we show that binding of large but not small DNAs by the C terminus of p53 negatively regulates sequence-specific DNA binding by the central domain. Four previously described mechanisms for activation of specific DNA binding operate by blocking negative regulation. Deletion of the C terminus of p53 activates specific DNA binding only in the presence of large DNA. Three activator molecules (a small nucleic acid, a monoclonal antibody against the p53 C terminus, and a C-terminal peptide of p53) stimulate sequence-specific DNA binding only in the presence of both large DNA and p53 with an intact C terminus. Our findings argue that interactions of the C terminus of p53 with genomic DNA in vivo would prevent p53 binding to specific promoters and that cellular mechanisms to block C-terminal DNA binding would be required.

Full Text

The Full Text of this article is available as a PDF (2.3 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abarzúa P., LoSardo J. E., Gubler M. L., Spathis R., Lu Y. A., Felix A., Neri A. Restoration of the transcription activation function to mutant p53 in human cancer cells. Oncogene. 1996 Dec 5;13(11):2477–2482. [PubMed] [Google Scholar]
  2. Bargonetti J., Manfredi J. J., Chen X., Marshak D. R., Prives C. A proteolytic fragment from the central region of p53 has marked sequence-specific DNA-binding activity when generated from wild-type but not from oncogenic mutant p53 protein. Genes Dev. 1993 Dec;7(12B):2565–2574. doi: 10.1101/gad.7.12b.2565. [DOI] [PubMed] [Google Scholar]
  3. Bayle J. H., Elenbaas B., Levine A. J. The carboxyl-terminal domain of the p53 protein regulates sequence-specific DNA binding through its nonspecific nucleic acid-binding activity. Proc Natl Acad Sci U S A. 1995 Jun 6;92(12):5729–5733. doi: 10.1073/pnas.92.12.5729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Demczuk S., Harbers M., Vennström B. Identification and analysis of all components of a gel retardation assay by combination with immunoblotting. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2574–2578. doi: 10.1073/pnas.90.7.2574. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Friedman P. N., Kern S. E., Vogelstein B., Prives C. Wild-type, but not mutant, human p53 proteins inhibit the replication activities of simian virus 40 large tumor antigen. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9275–9279. doi: 10.1073/pnas.87.23.9275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Funk W. D., Pak D. T., Karas R. H., Wright W. E., Shay J. W. A transcriptionally active DNA-binding site for human p53 protein complexes. Mol Cell Biol. 1992 Jun;12(6):2866–2871. doi: 10.1128/mcb.12.6.2866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gottlieb T. M., Oren M. p53 in growth control and neoplasia. Biochim Biophys Acta. 1996 Jun 7;1287(2-3):77–102. doi: 10.1016/0304-419x(95)00019-c. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. 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]
  11. Jayaraman J., Prives C. Activation of p53 sequence-specific DNA binding by short single strands of DNA requires the p53 C-terminus. Cell. 1995 Jun 30;81(7):1021–1029. doi: 10.1016/s0092-8674(05)80007-8. [DOI] [PubMed] [Google Scholar]
  12. Kern S. E., Kinzler K. W., Bruskin A., Jarosz D., Friedman P., Prives C., Vogelstein B. Identification of p53 as a sequence-specific DNA-binding protein. Science. 1991 Jun 21;252(5013):1708–1711. doi: 10.1126/science.2047879. [DOI] [PubMed] [Google Scholar]
  13. Ko L. J., Prives C. p53: puzzle and paradigm. Genes Dev. 1996 May 1;10(9):1054–1072. doi: 10.1101/gad.10.9.1054. [DOI] [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lee S., Elenbaas B., Levine A., Griffith J. p53 and its 14 kDa C-terminal domain recognize primary DNA damage in the form of insertion/deletion mismatches. Cell. 1995 Jun 30;81(7):1013–1020. doi: 10.1016/s0092-8674(05)80006-6. [DOI] [PubMed] [Google Scholar]
  16. Levine A. J. p53, the cellular gatekeeper for growth and division. Cell. 1997 Feb 7;88(3):323–331. doi: 10.1016/s0092-8674(00)81871-1. [DOI] [PubMed] [Google Scholar]
  17. Miller S. D., Moses K., Jayaraman L., Prives C. Complex formation between p53 and replication protein A inhibits the sequence-specific DNA binding of p53 and is regulated by single-stranded DNA. Mol Cell Biol. 1997 Apr;17(4):2194–2201. doi: 10.1128/mcb.17.4.2194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pavletich N. P., Chambers K. A., Pabo C. O. The DNA-binding domain of p53 contains the four conserved regions and the major mutation hot spots. Genes Dev. 1993 Dec;7(12B):2556–2564. doi: 10.1101/gad.7.12b.2556. [DOI] [PubMed] [Google Scholar]
  19. Reed M., Wang Y., Mayr G., Anderson M. E., Schwedes J. F., Tegtmeyer P. p53 domains: suppression, transformation, and transactivation. Gene Expr. 1993;3(1):95–107. [PMC free article] [PubMed] [Google Scholar]
  20. Reed M., Woelker B., Wang P., Wang Y., Anderson M. E., Tegtmeyer P. The C-terminal domain of p53 recognizes DNA damaged by ionizing radiation. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9455–9459. doi: 10.1073/pnas.92.21.9455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Selivanova G., Iotsova V., Kiseleva E., Ström M., Bakalkin G., Grafström R. C., Wiman K. G. The single-stranded DNA end binding site of p53 coincides with the C-terminal regulatory region. Nucleic Acids Res. 1996 Sep 15;24(18):3560–3567. doi: 10.1093/nar/24.18.3560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stenger J. E., Mayr G. A., Mann K., Tegtmeyer P. Formation of stable p53 homotetramers and multiples of tetramers. Mol Carcinog. 1992;5(2):102–106. doi: 10.1002/mc.2940050204. [DOI] [PubMed] [Google Scholar]
  23. Wang P., Reed M., Wang Y., Mayr G., Stenger J. E., Anderson M. E., Schwedes J. F., Tegtmeyer P. p53 domains: structure, oligomerization, and transformation. Mol Cell Biol. 1994 Aug;14(8):5182–5191. doi: 10.1128/mcb.14.8.5182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wang Y., Reed M., Wang P., Stenger J. E., Mayr G., Anderson M. E., Schwedes J. F., Tegtmeyer P. p53 domains: identification and characterization of two autonomous DNA-binding regions. Genes Dev. 1993 Dec;7(12B):2575–2586. doi: 10.1101/gad.7.12b.2575. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES