Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1989 Sep;63(9):3894–3901. doi: 10.1128/jvi.63.9.3894-3901.1989

Evolutionary conservation of the biochemical properties of p53: specific interaction of Xenopus laevis p53 with simian virus 40 large T antigen and mammalian heat shock proteins 70.

T Soussi 1, C Caron de Fromentel 1, H W Stürzbecher 1, S Ullrich 1, J Jenkins 1, P May 1
PMCID: PMC250985  PMID: 2668561

Abstract

We have investigated the biochemical properties of Xenopus laevis p53. With an in vitro binding assay, we can detect a specific association between X. laevis p53 and simian virus 40 large T antigen. Furthermore, X. laevis p53 expressed in monkey COS cells is stably associated with this viral antigen. Like mammalian p53, X. laevis p53 in complex with simian virus 40 large T antigen exhibits a 20-fold increase of its half-life. On the other hand, X. laevis p53 is unable to associate either in vivo or in vitro with adenovirus type 5 E1B 55-kilodalton protein. We show by an immunological technique that X. laevis p53 forms specific complexes with mammalian hsp72 and hsp73 heat shock proteins only at a temperature well above the optimal growth temperature for X. laevis. Our results suggest that the protein-binding properties of p53 are closely related to the functional activity of the protein.

Full text

PDF
3894

Images in this article

3896Image
on p.3896
3896Image
on p.3896
3897Image
on p.3897
3897Image
on p.3897
3898Image
on p.3898

Selected References

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

  1. Benoist C., Chambon P. Deletions covering the putative promoter region of early mRNAs of simian virus 40 do not abolish T-antigen expression. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3865–3869. doi: 10.1073/pnas.77.7.3865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Caron de Fromentel C., May-Levin F., Mouriesse H., Lemerle J., Chandrasekaran K., May P. Presence of circulating antibodies against cellular protein p53 in a notable proportion of children with B-cell lymphoma. Int J Cancer. 1987 Feb 15;39(2):185–189. doi: 10.1002/ijc.2910390211. [DOI] [PubMed] [Google Scholar]
  3. Caron de Fromentel C., Viron A., Puvion E., May P. SV40 large T-antigen and transformation related protein p53 are associated in situ with nuclear RNP structures containing hnRNA of transformed cells. Exp Cell Res. 1986 May;164(1):35–48. doi: 10.1016/0014-4827(86)90452-0. [DOI] [PubMed] [Google Scholar]
  4. Carroll R. B., Gurney E. G. Time-dependent maturation of the simian virus 40 large T antigen-p53 complex studied by using monoclonal antibodies. J Virol. 1982 Nov;44(2):565–573. doi: 10.1128/jvi.44.2.565-573.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clarke C. F., Cheng K., Frey A. B., Stein R., Hinds P. W., Levine A. J. Purification of complexes of nuclear oncogene p53 with rat and Escherichia coli heat shock proteins: in vitro dissociation of hsc70 and dnaK from murine p53 by ATP. Mol Cell Biol. 1988 Mar;8(3):1206–1215. doi: 10.1128/mcb.8.3.1206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crawford L. V., Pim D. C., Bulbrook R. D. Detection of antibodies against the cellular protein p53 in sera from patients with breast cancer. Int J Cancer. 1982 Oct 15;30(4):403–408. doi: 10.1002/ijc.2910300404. [DOI] [PubMed] [Google Scholar]
  7. DeLeo A. B., Jay G., Appella E., Dubois G. C., Law L. W., Old L. J. Detection of a transformation-related antigen in chemically induced sarcomas and other transformed cells of the mouse. Proc Natl Acad Sci U S A. 1979 May;76(5):2420–2424. doi: 10.1073/pnas.76.5.2420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Deppert W., Haug M. Evidence for free and metabolically stable p53 protein in nuclear subfractions of simian virus 40-transformed cells. Mol Cell Biol. 1986 Jun;6(6):2233–2240. doi: 10.1128/mcb.6.6.2233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deppert W., Haug M., Steinmayer T. Modulation of p53 protein expression during cellular transformation with simian virus 40. Mol Cell Biol. 1987 Dec;7(12):4453–4463. doi: 10.1128/mcb.7.12.4453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ehrhart J. C., Duthu A., Ullrich S., Appella E., May P. Specific interaction between a subset of the p53 protein family and heat shock proteins hsp72/hsc73 in a human osteosarcoma cell line. Oncogene. 1988 Nov;3(5):595–603. [PubMed] [Google Scholar]
  11. Eliyahu D., Raz A., Gruss P., Givol D., Oren M. Participation of p53 cellular tumour antigen in transformation of normal embryonic cells. Nature. 1984 Dec 13;312(5995):646–649. doi: 10.1038/312646a0. [DOI] [PubMed] [Google Scholar]
  12. Ernoult-Lange M., May P., Moreau P., May E. Simian virus 40 late promoter region able to initiate simian virus 40 early gene transcription in the absence of the simian virus 40 origin sequence. J Virol. 1984 Apr;50(1):163–173. doi: 10.1128/jvi.50.1.163-173.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Finlay C. A., Hinds P. W., Tan T. H., Eliyahu D., Oren M., Levine A. J. Activating mutations for transformation by p53 produce a gene product that forms an hsc70-p53 complex with an altered half-life. Mol Cell Biol. 1988 Feb;8(2):531–539. doi: 10.1128/mcb.8.2.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gannon J. V., Lane D. P. p53 and DNA polymerase alpha compete for binding to SV40 T antigen. Nature. 1987 Oct 1;329(6138):456–458. doi: 10.1038/329456a0. [DOI] [PubMed] [Google Scholar]
  15. Gaudray P., Rassoulzadegan M., Cuzin F. Expression of simian virus 40 early genes in transformed rat cells is correlated with maintenance of the transformed phenotype. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4987–4991. doi: 10.1073/pnas.75.10.4987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gluzman Y. SV40-transformed simian cells support the replication of early SV40 mutants. Cell. 1981 Jan;23(1):175–182. doi: 10.1016/0092-8674(81)90282-8. [DOI] [PubMed] [Google Scholar]
  17. Gronostajski R. M., Goldberg A. L., Pardee A. B. Energy requirement for degradation of tumor-associated protein p53. Mol Cell Biol. 1984 Mar;4(3):442–448. doi: 10.1128/mcb.4.3.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gurney E. G., Harrison R. O., Fenno J. Monoclonal antibodies against simian virus 40 T antigens: evidence for distinct sublcasses of large T antigen and for similarities among nonviral T antigens. J Virol. 1980 Jun;34(3):752–763. doi: 10.1128/jvi.34.3.752-763.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harlow E., Crawford L. V., Pim D. C., Williamson N. M. Monoclonal antibodies specific for simian virus 40 tumor antigens. J Virol. 1981 Sep;39(3):861–869. doi: 10.1128/jvi.39.3.861-869.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Harlow E., Williamson N. M., Ralston R., Helfman D. M., Adams T. E. Molecular cloning and in vitro expression of a cDNA clone for human cellular tumor antigen p53. Mol Cell Biol. 1985 Jul;5(7):1601–1610. doi: 10.1128/mcb.5.7.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Harrison T., Graham F., Williams J. Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology. 1977 Mar;77(1):319–329. doi: 10.1016/0042-6822(77)90428-7. [DOI] [PubMed] [Google Scholar]
  22. Hinds P. W., Finlay C. A., Frey A. B., Levine A. J. Immunological evidence for the association of p53 with a heat shock protein, hsc70, in p53-plus-ras-transformed cell lines. Mol Cell Biol. 1987 Aug;7(8):2863–2869. doi: 10.1128/mcb.7.8.2863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Jenkins J. R., Chumakov P., Addison C., Stürzbecher H. W., Wade-Evans A. Two distinct regions of the murine p53 primary amino acid sequence are implicated in stable complex formation with simian virus 40 T antigen. J Virol. 1988 Oct;62(10):3903–3906. doi: 10.1128/jvi.62.10.3903-3906.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Jenkins J. R., Rudge K., Chumakov P., Currie G. A. The cellular oncogene p53 can be activated by mutagenesis. 1985 Oct 31-Nov 6Nature. 317(6040):816–818. doi: 10.1038/317816a0. [DOI] [PubMed] [Google Scholar]
  25. Jenkins J. R., Rudge K., Currie G. A. Cellular immortalization by a cDNA clone encoding the transformation-associated phosphoprotein p53. Nature. 1984 Dec 13;312(5995):651–654. doi: 10.1038/312651a0. [DOI] [PubMed] [Google Scholar]
  26. Jenkins J. R., Rudge K., Redmond S., Wade-Evans A. Cloning and expression analysis of full length mouse cDNA sequences encoding the transformation associated protein p53. Nucleic Acids Res. 1984 Jul 25;12(14):5609–5626. doi: 10.1093/nar/12.14.5609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kress M., May E., Cassingena R., May P. Simian virus 40-transformed cells express new species of proteins precipitable by anti-simian virus 40 tumor serum. J Virol. 1979 Aug;31(2):472–483. doi: 10.1128/jvi.31.2.472-483.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Lane D. P., Crawford L. V. T antigen is bound to a host protein in SV40-transformed cells. Nature. 1979 Mar 15;278(5701):261–263. doi: 10.1038/278261a0. [DOI] [PubMed] [Google Scholar]
  31. Linzer D. I., Levine A. J. Characterization of a 54K dalton cellular SV40 tumor antigen present in SV40-transformed cells and uninfected embryonal carcinoma cells. Cell. 1979 May;17(1):43–52. doi: 10.1016/0092-8674(79)90293-9. [DOI] [PubMed] [Google Scholar]
  32. Manteuil S., Pages J., Stehelin D., Girard M. Replication of simian virus 40 deoxyribonucleic acid: analysis of the one-step growth cycle. J Virol. 1973 Jan;11(1):98–106. doi: 10.1128/jvi.11.1.98-106.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Matlashewski G., Lamb P., Pim D., Peacock J., Crawford L., Benchimol S. Isolation and characterization of a human p53 cDNA clone: expression of the human p53 gene. EMBO J. 1984 Dec 20;3(13):3257–3262. doi: 10.1002/j.1460-2075.1984.tb02287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. McCormick F., Clark R., Harlow E., Tjian R. SV40 T antigen binds specifically to a cellular 53 K protein in vitro. Nature. 1981 Jul 2;292(5818):63–65. doi: 10.1038/292063a0. [DOI] [PubMed] [Google Scholar]
  35. Melero J. A., Stitt D. T., Mangel W. F., Carroll R. B. Identification of new polypeptide species (48-55K) immunoprecipitable by antiserum to purified large T antigen and present in SV40-infected and -transformed cells. Virology. 1979 Mar;93(2):466–480. doi: 10.1016/0042-6822(79)90250-2. [DOI] [PubMed] [Google Scholar]
  36. Oren M., Levine A. J. Molecular cloning of a cDNA specific for the murine p53 cellular tumor antigen. Proc Natl Acad Sci U S A. 1983 Jan;80(1):56–59. doi: 10.1073/pnas.80.1.56. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Oren M., Maltzman W., Levine A. J. Post-translational regulation of the 54K cellular tumor antigen in normal and transformed cells. Mol Cell Biol. 1981 Feb;1(2):101–110. doi: 10.1128/mcb.1.2.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Parada L. F., Land H., Weinberg R. A., Wolf D., Rotter V. Cooperation between gene encoding p53 tumour antigen and ras in cellular transformation. Nature. 1984 Dec 13;312(5995):649–651. doi: 10.1038/312649a0. [DOI] [PubMed] [Google Scholar]
  39. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  40. Pennica D., Goeddel D. V., Hayflick J. S., Reich N. C., Anderson C. W., Levine A. J. The amino acid sequence of murine p53 determined from a c-DNA clone. Virology. 1984 Apr 30;134(2):477–482. doi: 10.1016/0042-6822(84)90316-7. [DOI] [PubMed] [Google Scholar]
  41. Pinhasi-Kimhi O., Michalovitz D., Ben-Zeev A., Oren M. Specific interaction between the p53 cellular tumour antigen and major heat shock proteins. Nature. 1986 Mar 13;320(6058):182–184. doi: 10.1038/320182a0. [DOI] [PubMed] [Google Scholar]
  42. Pinhasi O., Oren M. Expression of the mouse p53 cellular tumor antigen in monkey cells. Mol Cell Biol. 1984 Oct;4(10):2180–2186. doi: 10.1128/mcb.4.10.2180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Reich N. C., Oren M., Levine A. J. Two distinct mechanisms regulate the levels of a cellular tumor antigen, p53. Mol Cell Biol. 1983 Dec;3(12):2143–2150. doi: 10.1128/mcb.3.12.2143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rotter V., Witte O. N., Coffman R., Baltimore D. Abelson murine leukemia virus-induced tumors elicit antibodies against a host cell protein, P50. J Virol. 1980 Nov;36(2):547–555. doi: 10.1128/jvi.36.2.547-555.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Ruscetti S. K., Scolnick E. M. Expression of a transformation-related protein (p53) in the malignant stage of Friend virus-induced diseases. J Virol. 1983 Jun;46(3):1022–1026. doi: 10.1128/jvi.46.3.1022-1026.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sarnow P., Ho Y. S., Williams J., Levine A. J. Adenovirus E1b-58kd tumor antigen and SV40 large tumor antigen are physically associated with the same 54 kd cellular protein in transformed cells. Cell. 1982 Feb;28(2):387–394. doi: 10.1016/0092-8674(82)90356-7. [DOI] [PubMed] [Google Scholar]
  47. Soussi T., Bègue A., Kress M., Stehelin D., May P. Nucleotide sequence of a cDNA encoding the chicken p53 nuclear oncoprotein. Nucleic Acids Res. 1988 Dec 9;16(23):11383–11383. doi: 10.1093/nar/16.23.11383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Soussi T., Caron de Fromentel C., Breugnot C., May E. Nucleotide sequence of a cDNA encoding the rat p53 nuclear oncoprotein. Nucleic Acids Res. 1988 Dec 9;16(23):11384–11384. doi: 10.1093/nar/16.23.11384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Soussi T., Caron de Fromentel C., Méchali M., May P., Kress M. Cloning and characterization of a cDNA from Xenopus laevis coding for a protein homologous to human and murine p53. Oncogene. 1987 Mar;1(1):71–78. [PubMed] [Google Scholar]
  50. Stürzbecher H. W., Addison C., Jenkins J. R. Characterization of mutant p53-hsp72/73 protein-protein complexes by transient expression in monkey COS cells. Mol Cell Biol. 1988 Sep;8(9):3740–3747. doi: 10.1128/mcb.8.9.3740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Stürzbecher H. W., Chumakov P., Welch W. J., Jenkins J. R. Mutant p53 proteins bind hsp 72/73 cellular heat shock-related proteins in SV40-transformed monkey cells. Oncogene. 1987 May;1(2):201–211. [PubMed] [Google Scholar]
  52. Tan T. H., Wallis J., Levine A. J. Identification of the p53 protein domain involved in formation of the simian virus 40 large T-antigen-p53 protein complex. J Virol. 1986 Sep;59(3):574–583. doi: 10.1128/jvi.59.3.574-583.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wolf D., Laver-Rudich Z., Rotter V. In vitro expression of human p53 cDNA clones and characterization of the cloned human p53 gene. Mol Cell Biol. 1985 Aug;5(8):1887–1893. doi: 10.1128/mcb.5.8.1887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Zakut-Houri R., Bienz-Tadmor B., Givol D., Oren M. Human p53 cellular tumor antigen: cDNA sequence and expression in COS cells. EMBO J. 1985 May;4(5):1251–1255. doi: 10.1002/j.1460-2075.1985.tb03768.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zakut-Houri R., Oren M., Bienz B., Lavie V., Hazum S., Givol D. A single gene and a pseudogene for the cellular tumour antigen p53. Nature. 1983 Dec 8;306(5943):594–597. doi: 10.1038/306594a0. [DOI] [PubMed] [Google Scholar]
  56. Zantema A., Schrier P. I., Davis-Olivier A., van Laar T., Vaessen R. T., van der EB A. J. Adenovirus serotype determines association and localization of the large E1B tumor antigen with cellular tumor antigen p53 in transformed cells. Mol Cell Biol. 1985 Nov;5(11):3084–3091. doi: 10.1128/mcb.5.11.3084. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES