Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1980 Aug;35(2):320–330. doi: 10.1128/jvi.35.2.320-330.1980

Association of simian virus 40 T antigen with replicating nucleoprotein complexes of simian virus 40.

M Segawa, S Sugano, N Yamaguchi
PMCID: PMC288816  PMID: 6255173

Abstract

An immunoprecipitation assay was established for simian virus 40 T-antigen-bound nucleoprotein complexes by means of precipitation with sera from hamsters bearing simian virus 40-induced tumors. About 80% of simian virus 40 replicating nucleoprotein complexes in various stages of replication were immunoprecipitated. In contrast, less than 21% of mature nucleoprotein complexes were immunoprecipitated. Pulse-chase experiments showed that T antigen was lost from most of the nucleoprotein complexes concurrently with completion of DNA replication. T antigen induced by dl-940, a mutant with a deletion in the region coding for small T antigen, was also associated with most of the replicating nucleoprotein complexes. Once bound with replicating nucleoprotein complexes at the permissive temperature, thermolabile T antigen induced by tsA900 remained associated with the complexes during elongation of the replicating DNA chain at the restrictive temperature. These results suggest that simian virus 40 T antigen (probably large T antigen) associates with nucleoprotein complexes at or before initiation of DNA replication and that the majority of the T antigen dissociates from the nucleoprotein complexes simultaneously with completion of DNA replication.

Full text

PDF
321

Selected References

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

  1. Baumgartner I., Kuhn C., Fanning E. Identification and characterization of fast-sedimenting SV40 nucleoprotein complexes. Virology. 1979 Jul 15;96(1):54–63. doi: 10.1016/0042-6822(79)90172-7. [DOI] [PubMed] [Google Scholar]
  2. Birkenmeier E. H., Chiu N., Radonovich M. F., May E., Salzman N. P. Regulation of simian virus 40 early and late gene transcription without viral DNA replication. J Virol. 1979 Mar;29(3):983–989. doi: 10.1128/jvi.29.3.983-989.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouck N., Beales N., Shenk T., Berg P., di Mayorca G. New region of the simian virus 40 genome required for efficient viral transformation. Proc Natl Acad Sci U S A. 1978 May;75(5):2473–2477. doi: 10.1073/pnas.75.5.2473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Carroll R. B., Hager L., Dulbecco R. Simian virus 40 T antigen binds to DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3754–3757. doi: 10.1073/pnas.71.9.3754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Chou J. Y., Martin R. G. DNA infectivity and the induction of host DNA synthesis with temperature-sensitive mutants of simian virus 40. J Virol. 1975 Jan;15(1):145–150. doi: 10.1128/jvi.15.1.145-150.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cowan K., Tegtmeyer P., Anthony D. D. Relationship of replication and transcription of Simian Virus 40 DNA. Proc Natl Acad Sci U S A. 1973 Jul;70(7):1927–1930. doi: 10.1073/pnas.70.7.1927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Crawford L. V., Cole C. N., Smith A. E., Paucha E., Tegtmeyer P., Rundell K., Berg P. Organization and expression of early genes of simian virus 40. Proc Natl Acad Sci U S A. 1978 Jan;75(1):117–121. doi: 10.1073/pnas.75.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edenberg H. J., Anderson S., DePamphilis M. L. Involvement of DNA polymerase alpha in simian virus 40 DNA replication. J Biol Chem. 1978 May 10;253(9):3273–3280. [PubMed] [Google Scholar]
  10. Edwards C. A., Khoury G., Martin R. G. Phosphorylation of T-antigen and control T-antigen expression in cells transformed by wild-type and tsA mutants of simian virus 40. J Virol. 1979 Feb;29(2):753–762. doi: 10.1128/jvi.29.2.753-762.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fareed G. C., Davoli D. Molecular biology of papovaviruses. Annu Rev Biochem. 1977;46:471–522. doi: 10.1146/annurev.bi.46.070177.002351. [DOI] [PubMed] [Google Scholar]
  12. Ferdinand F. J., Brown M., Khoury G. Characterization of early simian virus 40 transcriptional complexes: late transcription in the absence of detectable DNA replication. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5443–5447. doi: 10.1073/pnas.74.12.5443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fernandez-Munoz R., Coca-Prados M., Hsu M. T. Intracellular forms of simian virus 40 nucleoprotein complexes. I. Methods of isolation and characterization in CV-1 cells. J Virol. 1979 Feb;29(2):612–623. doi: 10.1128/jvi.29.2.612-623.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Feunteun J., Kress M., Gardes M., Monier R. Viable deletion mutants in the simian virus 40 early region. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4455–4459. doi: 10.1073/pnas.75.9.4455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Garber E. A., Seidman M. M., Levine A. J. The detection and characterization of multiple forms of SV40 nucleoprotein complexes. Virology. 1978 Oct 15;90(2):305–316. doi: 10.1016/0042-6822(78)90315-x. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Green M. H., Brooks T. L. Recently replicated simian virus 40 DNA is a preferential template for transcription and replication. J Virol. 1978 May;26(2):325–334. doi: 10.1128/jvi.26.2.325-334.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. HOGGAN M. D., ROWE W. P., BLACK P. H., HUEBNER R. J. PRODUCTION OF "TUMOR-SPECIFIC" ANTIGENS BY ONCOGENIC VIRUSES DURING ACUTE CYTOLYTIC INFECTIONS. Proc Natl Acad Sci U S A. 1965 Jan;53:12–19. doi: 10.1073/pnas.53.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  20. Jerkofsky M. Enhancement of the replication of human adenovirus in simian cells by a series of temperature-sensitive mutants of simian virus 40. Virology. 1975 Jun;65(2):579–582. doi: 10.1016/0042-6822(75)90063-x. [DOI] [PubMed] [Google Scholar]
  21. Jessel D., Landau T., Hudson J., Lalor T., Tenen D., Livingston D. M. Identification of regions of the SV40 genome which contain preferred SV40 T antigen-binding sites. Cell. 1976 Aug;8(4):535–545. doi: 10.1016/0092-8674(76)90222-1. [DOI] [PubMed] [Google Scholar]
  22. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  23. 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]
  24. Levine A. J., Kang H. S., Billheimer F. E. DNA replication in SV40 infected cells. I. Analysis of replicating SV40 DNA. J Mol Biol. 1970 Jun 14;50(2):549–568. doi: 10.1016/0022-2836(70)90211-1. [DOI] [PubMed] [Google Scholar]
  25. Lewis A. M., Jr, Rowe W. P. Studies on nondefective adenovirus-simian virus 40 hybrid viruses. I. A newly characterized simian virus 40 antigen induced by the Ad2+ND 1 virus. J Virol. 1971 Feb;7(2):189–197. doi: 10.1128/jvi.7.2.189-197.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Mann K., Hunter T. Association of simian virus 40 T antigen with simian virus 40 nucleoprotein complexes. J Virol. 1979 Jan;29(1):232–241. doi: 10.1128/jvi.29.1.232-241.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Martin R. G., Setlow V. P., Edwards C. A., Vembu D. The roles of the simian virus 40 tumor antigens in transformation of Chinese hamster lung cells. Cell. 1979 Jul;17(3):635–643. doi: 10.1016/0092-8674(79)90271-x. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Osborn M., Weber K. SV40: T antigen, the A function and transformation. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):267–276. doi: 10.1101/sqb.1974.039.01.035. [DOI] [PubMed] [Google Scholar]
  31. Otto B., Fanning E. DNA polymerase alpha is associated with replicating SV40 nucleoprotein complexes. Nucleic Acids Res. 1978 May;5(5):1715–1728. doi: 10.1093/nar/5.5.1715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Persico-DiLauro M., Martin R. G., Livingston D. M. Interaction of Simian Virus 40 chromatin with Simian Virus 40 T-antigen. J Virol. 1977 Nov;24(2):451–460. doi: 10.1128/jvi.24.2.451-460.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Potter C. W., McLaughlin B. C., Oxford J. S. Simian virus 40-induced T and tumor antigens. J Virol. 1969 Nov;4(5):574–579. doi: 10.1128/jvi.4.5.574-579.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Prives C., Gilboa E., Revel M., Winocour E. Cell-free translation of simian virus 40 early messenger RNA coding for viral T-antigen. Proc Natl Acad Sci U S A. 1977 Feb;74(2):457–461. doi: 10.1073/pnas.74.2.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Reed S. I., Ferguson J., Davis R. W., Stark G. R. T antigen binds to simian virus 40 DNA at the origin of replication. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1605–1609. doi: 10.1073/pnas.72.4.1605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Robb J. A. Identification of simian virus 40 tumor and U antigens. Proc Natl Acad Sci U S A. 1977 Feb;74(2):447–451. doi: 10.1073/pnas.74.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rundell K., Collins J. K., Tegtmeyer P., Ozer H. L., Lai C. J., Nathans D. Identification of simian virus 40 protein A. J Virol. 1977 Feb;21(2):636–646. doi: 10.1128/jvi.21.2.636-646.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. SEVER J. L. Application of a microtechnique to viral serological investigations. J Immunol. 1962 Mar;88:320–329. [PubMed] [Google Scholar]
  40. Segawa K., Yamaguchi N., Oda K. Simian virus 40 gene A regulates the association between a highly phosphorylated protein and chromatin and ribosomes in simian virus 40-transformed cells. J Virol. 1977 Jun;22(3):679–693. doi: 10.1128/jvi.22.3.679-693.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Seidman M. M., Salzman N. P. Late replicative intermediates are accumulated during simian virus 40 DNA replication in vivo and in vitro. J Virol. 1979 May;30(2):600–609. doi: 10.1128/jvi.30.2.600-609.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Shenk T. E., Carbon J., Berg P. Construction and analysis of viable deletion mutants of simian virus 40. J Virol. 1976 May;18(2):664–671. doi: 10.1128/jvi.18.2.664-671.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sleigh M. J., Topp W. C., Hanich R., Sambrook J. F. Mutants of SV40 with an altered small t protein are reduced in their ability to transform cells. Cell. 1978 May;14(1):79–88. doi: 10.1016/0092-8674(78)90303-3. [DOI] [PubMed] [Google Scholar]
  44. Tapper D. P., DePamphilis M. L. Discontinuous DNA replication: accumulation of Simian virus 40 DNA at specific stages in its replication. J Mol Biol. 1978 Apr 15;120(3):401–422. doi: 10.1016/0022-2836(78)90427-8. [DOI] [PubMed] [Google Scholar]
  45. Tegtmeyer P., Macasaet F. Simian virus 40 deoxyribonucleic acid synthesis: analysis by gel electrophoresis. J Virol. 1972 Oct;10(4):599–604. doi: 10.1128/jvi.10.4.599-604.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Tegtmeyer P., Ozer H. L. Temperature-sensitive mutants of simian virus 40: infection of permissive cells. J Virol. 1971 Oct;8(4):516–524. doi: 10.1128/jvi.8.4.516-524.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Tenen D. G., Baygell P., Livingston D. M. Thermolabile T (tumor) antigen from cells transformed by a temperature-sensitive mutant of simian virus 40. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4351–4355. doi: 10.1073/pnas.72.11.4351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Tjian R. The binding site on SV40 DNA for a T antigen-related protein. Cell. 1978 Jan;13(1):165–179. doi: 10.1016/0092-8674(78)90147-2. [DOI] [PubMed] [Google Scholar]
  51. Yamaguchi N., Kuchino T. Temperature-sensitive mutants of simian virus 40 selected by transforming ability. J Virol. 1975 Jun;15(6):1297–1301. doi: 10.1128/jvi.15.6.1297-1301.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Yang Y. C., Hearing P., Rundell K. Cellular proteins associated with simian virus 40 early gene products in newly infected cells. J Virol. 1979 Oct;32(1):147–154. doi: 10.1128/jvi.32.1.147-154.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES