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. 1984 Aug;51(2):376–383. doi: 10.1128/jvi.51.2.376-383.1984

Antigenic structure of simian virus 40 large tumor antigen and association with cellular protein p53 on the surfaces of simian virus 40-infected and -transformed cells.

M Santos, J S Butel
PMCID: PMC254448  PMID: 6205166

Abstract

The antigenic structure of simian virus 40 (SV40) large tumor antigen (T-ag) in the plasma membranes of SV40-transformed mouse cells and SV40-infected monkey cells was characterized as a step toward defining possible biological function(s). Wild-type SV40, as well as a deletion mutant of SV40 (dl1263) which codes for a truncated T-ag with an altered carboxy terminus, was used to infect permissive cells. Members of a series of monoclonal antibodies directed against antigenic determinants on either the amino or the carboxy terminus of the T-ag polypeptide were able to precipitate surface T-ag (as well as nuclear T-ag) from both SV40-transformed and SV40-infected cells. Cellular protein p53 was coprecipitated with T-ag by all T-ag-reactive reagents from the surface and nucleus of SV40-transformed cells. In contrast, T-ag, but not T-ag-p53 complex, was recovered from the surface of SV40-infected cells. These results confirm that nuclear T-ag and surface T-ag are highly related molecules and that a complex of SV40 T-ag and p53 is present at the surface of SV40-transformed cells. Detectable levels of such a complex do not appear to be present on SV40-infected cells. Both the carboxy and amino termini of T-ag are exposed on the surfaces of SV40-transformed and -infected cells. The possible relevance of the presence of a T-ag-p53 complex on the surface of SV40-transformed cells and its absence from SV40-infected cells is considered.

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  1. Alwine J. C., Reed S. I., Stark G. R. Characterization of the autoregulation of simian virus 40 gene A. J Virol. 1977 Oct;24(1):22–27. doi: 10.1128/jvi.24.1.22-27.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brugge J. S., Butel J. S. Role of simian virus 40 gene A function in maintenance of transformation. J Virol. 1975 Mar;15(3):619–635. doi: 10.1128/jvi.15.3.619-635.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Butel J. S., Soule H. R. Role of the simian virus 40 gene A product in regulation of DNA synthesis in transformed cells. J Virol. 1978 Jun;26(3):584–594. doi: 10.1128/jvi.26.3.584-594.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Butel J. S., Tevethia S. S., Melnick J. L. Oncogenicity and cell transformation by papovavirus SV40: the role of the viral genome. Adv Cancer Res. 1972;15:1–55. doi: 10.1016/s0065-230x(08)60371-1. [DOI] [PubMed] [Google Scholar]
  5. Carmichael G. G., Schaffhausen B. S., Dorsky D. I., Oliver D. B., Benjamin T. L. Carboxy terminus of polyoma middle-sized tumor antigen is required for attachment to membranes, associated protein kinase activities, and cell transformation. Proc Natl Acad Sci U S A. 1982 Jun;79(11):3579–3583. doi: 10.1073/pnas.79.11.3579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chandrasekaran K., Winterbourne D. J., Luborsky S. W., Mora P. T. Surface proteins of simian-virus-40-transformed cells. Int J Cancer. 1981 Mar 15;27(3):397–407. doi: 10.1002/ijc.2910270320. [DOI] [PubMed] [Google Scholar]
  7. Chou J. Y., Avila J., Martin R. G. Viral DNA synthesis in cells infected by temperature-sensitive mutants of simian virus 40. J Virol. 1974 Jul;14(1):116–124. doi: 10.1128/jvi.14.1.116-124.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Christensen J. B., Brockman W. W. Effects of large and small T antigens on DNA synthesis and cell division in simian virus 40-transformed BALB/c 3T3 cells. J Virol. 1982 Nov;44(2):574–585. doi: 10.1128/jvi.44.2.574-585.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clark R., Lane D. P., Tjian R. Use of monoclonal antibodies as probes of simian virus 40 T antigen ATPase activity. J Biol Chem. 1981 Nov 25;256(22):11854–11858. [PubMed] [Google Scholar]
  10. Cole C. N., Crawford L. V., Berg P. Simian virus 40 mutants with deletions at the 3' end of the early region are defective in adenovirus helper function. J Virol. 1979 Jun;30(3):683–691. doi: 10.1128/jvi.30.3.683-691.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cole C. N., Landers T., Goff S. P., Manteuil-Brutlag S., Berg P. Physical and genetic characterization of deletion mutants of simian virus 40 constructed in vitro. J Virol. 1977 Oct;24(1):277–294. doi: 10.1128/jvi.24.1.277-294.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Crawford L., Harlow E. Uniform nomenclature for monoclonal antibodies directed against virus-coded proteins of simian virus 40 and polyoma virus. J Virol. 1982 Feb;41(2):709–709. doi: 10.1128/jvi.41.2.709-709.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Crawford L., Leppard K., Lane D., Harlow E. Cellular proteins reactive with monoclonal antibodies directed against simian virus 40 T-antigen. J Virol. 1982 May;42(2):612–620. doi: 10.1128/jvi.42.2.612-620.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Deppert W., Gurney E. G., Harrison R. O. Monoclonal antibodies against simian virus 40 tumor antigens: analysis of antigenic binding sites, using adenovirus type 2-simian virus 40 hybrid viruses. J Virol. 1981 Jan;37(1):478–482. doi: 10.1128/jvi.37.1.478-482.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Deppert W., Hanke K., Henning R. Simian virus 40 T-antigen-related cell surface antigen: serological demonstration on simian virus 40-transformed monolayer cells in situ. J Virol. 1980 Aug;35(2):505–518. doi: 10.1128/jvi.35.2.505-518.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Deppert W., Pates R. Cell surface location of simian virus 40-specific proteins on HeLa cells infected with adenovirus type 2-simian virus 40 hybrid viruses Ad2+ND1 and Ad2+ND2. J Virol. 1979 Aug;31(2):522–536. doi: 10.1128/jvi.31.2.522-536.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Deppert W., Walter G. Domains of simian virus 40 large T-antigen exposed on the cell surface. Virology. 1982 Oct 15;122(1):56–70. doi: 10.1016/0042-6822(82)90377-4. [DOI] [PubMed] [Google Scholar]
  18. Deppert W., Walter G. Simian virus to (SV40) tumor-specific proteins in nucleus and plasma membrane of HeLa cells infected by adenovirus 2-SV40 hybrid virus Ad2+ND2. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2505–2509. doi: 10.1073/pnas.73.7.2505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Floros J., Jonak G., Galanti N., Baserga R. Induction of cell DNA replication in G1-specific ts mutants by microinjection of SV40 DNA. Exp Cell Res. 1981 Mar;132(1):215–223. doi: 10.1016/0014-4827(81)90097-5. [DOI] [PubMed] [Google Scholar]
  20. Galanti N., Jonak G. J., Soprano K. J., Floros J., Kaczmarek L., Weissman S., Reddy V. B., Tilghman S. M., Baserga R. Characterization and biological activity of cloned simian virus 40 DNA fragments. J Biol Chem. 1981 Jun 25;256(12):6469–6474. [PubMed] [Google Scholar]
  21. Graessmann M., Graessman A. "Early" simian-virus-40-specific RNA contains information for tumor antigen formation and chromatin replication. Proc Natl Acad Sci U S A. 1976 Feb;73(2):366–370. doi: 10.1073/pnas.73.2.366. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Harlow E., Pim D. C., Crawford L. V. Complex of simian virus 40 large-T antigen and host 53,000-molecular-weight protein in monkey cells. J Virol. 1981 Feb;37(2):564–573. doi: 10.1128/jvi.37.2.564-573.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Henning R., Lange-Mutschler J., Deppert W. SV40-transformed cells express SV40 T antigen-related antigens on the cell surface. Virology. 1981 Jan 30;108(2):325–337. doi: 10.1016/0042-6822(81)90441-4. [DOI] [PubMed] [Google Scholar]
  26. Ide T., Whelly S., Baserga R. Stimulation of RNA synthesis in isolated nuclei by partially purified preparations of simian virus 40 T-antigen. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3189–3192. doi: 10.1073/pnas.74.8.3189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ismail A., Baumann E. A., Hand R. Cell surface T antigen in cells infected with simian virus 40 or an adenovirus-simian virus 40 hybrid, Ad2+D2. J Virol. 1981 Nov;40(2):615–619. doi: 10.1128/jvi.40.2.615-619.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kimura G., Itagaki A. Initiation and maintenance of cell transformation by simian virus 40: a viral genetic property. Proc Natl Acad Sci U S A. 1975 Feb;72(2):673–677. doi: 10.1073/pnas.72.2.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Kit S., Kurimura T., Dubbs D. R. Transplantable mouse tumor line induced by injection of SV40-transformed mouse kidney cells. Int J Cancer. 1969 Jul 15;4(4):384–392. doi: 10.1002/ijc.2910040403. [DOI] [PubMed] [Google Scholar]
  31. Klockmann U., Deppert W. Acylation: a new post-translational modification specific for plasma membrane-associated simian virus 40 large T-antigen. FEBS Lett. 1983 Jan 24;151(2):257–259. doi: 10.1016/0014-5793(83)80081-7. [DOI] [PubMed] [Google Scholar]
  32. Lanford R. E., Butel J. S. Antigenic relationship of SV40 early proteins to purified large T polypeptide. Virology. 1979 Sep;97(2):295–306. doi: 10.1016/0042-6822(79)90341-6. [DOI] [PubMed] [Google Scholar]
  33. Lanford R. E., Butel J. S. Intracellular transport of SV40 large tumor antigen: a mutation which abolishes migration to the nucleus does not prevent association with the cell surface. Virology. 1982 May;119(1):169–184. doi: 10.1016/0042-6822(82)90074-5. [DOI] [PubMed] [Google Scholar]
  34. Lange-Mutschler J., Deppert W., Hanke K., Henning R. Detection of simian virus 40 T-antigen-related antigens by a 125I-protein A-binding assay and by immunofluorescence microscopy on the surface of SV40-transformed monolayer cells. J Gen Virol. 1981 Feb;52(Pt 2):301–312. doi: 10.1099/0022-1317-52-2-301. [DOI] [PubMed] [Google Scholar]
  35. Lebowitz P., Weissman S. M. Organization and transcription of the simian virus 40 genome. Curr Top Microbiol Immunol. 1979;87:43–172. doi: 10.1007/978-3-642-67344-3_3. [DOI] [PubMed] [Google Scholar]
  36. Luborsky S. W., Chandrasekaran K. Subcellular distribution of simian virus 40 T antigen species in various cell lines: the 56K protein. Int J Cancer. 1980 Apr 15;25(4):517–527. doi: 10.1002/ijc.2910250414. [DOI] [PubMed] [Google Scholar]
  37. Martin R. G., Chou J. Y. Simian virus 40 functions required for the establishment and maintenance of malignant transformation. J Virol. 1975 Mar;15(3):599–612. doi: 10.1128/jvi.15.3.599-612.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nicolson G. L. Trans-membrane control of the receptors on normal and tumor cells. II. Surface changes associated with transformation and malignancy. Biochim Biophys Acta. 1976 Apr 30;458(1):1–72. doi: 10.1016/0304-419x(76)90014-7. [DOI] [PubMed] [Google Scholar]
  39. Noonan C. A., Brugge J. S., Butel J. S. Characterization of simian cells tranformed by temperature-sensitive mutants of simian virus 40. J Virol. 1976 Jun;18(3):1106–1119. doi: 10.1128/jvi.18.3.1106-1119.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Noonan C. A., Butel J. S. Temperature-sensitive mutants of simian virus 40. I. Isolation aand preliminary characterization of B/C gene mutants. Intervirology. 1978;10(3):181–195. doi: 10.1159/000148982. [DOI] [PubMed] [Google Scholar]
  41. Osborn M., Weber K. Simian virus 40 gene A function and maintenance of transformation. J Virol. 1975 Mar;15(3):636–644. doi: 10.1128/jvi.15.3.636-644.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rapp F., Pauluzzi S., Butel J. S. Variation in properties of plaque progeny of PARA (defective simian papovavirus 40)-adenovirus 7. J Virol. 1969 Nov;4(5):626–631. doi: 10.1128/jvi.4.5.626-631.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Rio D., Robbins A., Myers R., Tjian R. Regulation of simian virus 40 early transcription in vitro by a purified tumor antigen. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5706–5710. doi: 10.1073/pnas.77.10.5706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Santos M., Butel J. S. Association of SV40 large tumor antigen and cellular proteins on the surface of SV40-transformed mouse cells. Virology. 1982 Jul 15;120(1):1–17. doi: 10.1016/0042-6822(82)90002-2. [DOI] [PubMed] [Google Scholar]
  46. Santos M., Butel J. S. Detection of a complex of SV40 large tumor antigen and 53K cellular protein on the surface of SV40-transformed mouse cells. J Cell Biochem. 1982;19(2):127–144. doi: 10.1002/jcb.240190204. [DOI] [PubMed] [Google Scholar]
  47. Santos M., Butel J. S. Dynamic nature of the association of large tumor antigen and p53 cellular protein with the surfaces of simian virus 40-transformed cells. J Virol. 1984 Jan;49(1):50–56. doi: 10.1128/jvi.49.1.50-56.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Schmidt-Ullrich R., Thompson W. S., Kahn S. J., Monroe M. T., Wallach D. F. Simian virus 40 (SV40)-specific isoelectric point-4.7--94,000-Mr membrane glycoprotein: major peptide homology exhibited with the nuclear and membrane-associated 94,000-Mr SV40 T-antigen in hamsters. J Natl Cancer Inst. 1982 Oct;69(4):839–849. [PubMed] [Google Scholar]
  49. Soprano K. J., Dev V. G., Croce C. M., Baserga R. Reactivation of silent rRNA genes by simian virus 40 in human-mouse hybrid cells. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3885–3889. doi: 10.1073/pnas.76.8.3885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Soprano K. J., Galanti N., Jonak G. J., McKercher S., Pipas J. M., Peden K. W., Baserga R. Mutational analysis of simian virus 40 T antigen: stimulation of cellular DNA synthesis and activation of rRNA genes by mutants with deletions in the T-antigen gene. Mol Cell Biol. 1983 Feb;3(2):214–219. doi: 10.1128/mcb.3.2.214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Soprano K. J., Jonak G. J., Galanti N., Floros J., Baserga R. Identification of an SV40 DNA sequence related to the reactivation of silent rRNA genes in human greater than mouse hybrid cells. Virology. 1981 Feb;109(1):127–136. doi: 10.1016/0042-6822(81)90477-3. [DOI] [PubMed] [Google Scholar]
  52. Soprano K. J., Rossini M., Croce C., Baserga R. The role of large T antigen in simian virus 40-induced reactivation of silent rRNA genes in human-mouse hybrid cells. Virology. 1980 Apr 30;102(2):317–326. doi: 10.1016/0042-6822(80)90099-9. [DOI] [PubMed] [Google Scholar]
  53. Soule H. R., Butel J. S. Subcellular Localization of simian virus 40 large tumor antigen. J Virol. 1979 May;30(2):523–532. doi: 10.1128/jvi.30.2.523-532.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Soule H. R., Lanford R. E., Butel J. S. Antigenic and immunogenic characteristics of nuclear and membrane-associated simian virus 40 tumor antigen. J Virol. 1980 Feb;33(2):887–901. doi: 10.1128/jvi.33.2.887-901.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Soule H. R., Lanford R. E., Butel J. S. Detection of simian virus 40 surface-associated large tumor antigen by enzyme-catalyzed radioiodination. Int J Cancer. 1982 Mar 15;29(3):337–344. doi: 10.1002/ijc.2910290318. [DOI] [PubMed] [Google Scholar]
  56. Sveda M. M., Markoff L. J., Lai C. J. Cell surface expression of the influenza virus hemagglutinin requires the hydrophobic carboxy-terminal sequences. Cell. 1982 Sep;30(2):649–656. doi: 10.1016/0092-8674(82)90261-6. [DOI] [PubMed] [Google Scholar]
  57. Tegtmeyer P. Function of simian virus 40 gene A in transforming infection. J Virol. 1975 Mar;15(3):613–618. doi: 10.1128/jvi.15.3.613-618.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Tegtmeyer P. Simian virus 40 deoxyribonucleic acid synthesis: the viral replicon. J Virol. 1972 Oct;10(4):591–598. doi: 10.1128/jvi.10.4.591-598.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Tevethia S. S., Tevethia M. J., Lewis A. J., Reddy V. B., Weissman S. M. Biology of simian virus 40 (SV40) transplantation antigen (TrAg). IX. Analysis of TrAg in mouse cells synthesizing truncated SV40 large T antigen. Virology. 1983 Jul 30;128(2):319–330. doi: 10.1016/0042-6822(83)90259-3. [DOI] [PubMed] [Google Scholar]
  60. Tjian R., Fey G., Graessmann A. Biological activity of purified simian virus 40 T antigen proteins. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1279–1283. doi: 10.1073/pnas.75.3.1279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Van Heuverswyn H., Cole C., Berg P., Fiers W. Nucleotide sequence analysis of two simian virus 40 mutants with deletions in the region coding for the carboxyl terminus of the T antigen. J Virol. 1979 Jun;30(3):936–941. doi: 10.1128/jvi.30.3.936-941.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Weil R. Viral 'tumor antigens': A novel type of mammalian regulator protein. Biochim Biophys Acta. 1978 Nov 17;516(3):301–388. doi: 10.1016/0304-419x(78)90012-4. [DOI] [PubMed] [Google Scholar]
  63. Whelly S., Ide T., Baserga R. Stimulation of RNA synthesis in isolated nucleoli by preparations of simian virus 40 T antigen. Virology. 1978 Jul 1;88(1):82–91. doi: 10.1016/0042-6822(78)90112-5. [DOI] [PubMed] [Google Scholar]
  64. Zarling J. M., Tevethia S. S. Transplantation immunity to simian virus 40-transformed cells in tumor-bearing mice. I. Development of cellular immunity to simian virus 40 tumor-specific transplantation antigens during tumorigenesis by transplanted cells. J Natl Cancer Inst. 1973 Jan;50(1):137–147. doi: 10.1093/jnci/50.1.137. [DOI] [PubMed] [Google Scholar]

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