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. 1980 Apr;34(1):213–224. doi: 10.1128/jvi.34.1.213-224.1980

Association of a murine 53,000-dalton phosphoprotein with simian virus 40 large-T antigen in transformed cells.

F McCormick, E Harlow
PMCID: PMC288687  PMID: 6246255

Abstract

Serum raised against a mouse 53,000-dalton (53K) phosphoprotein precipitates both the 53K immunogen and simian virus 40 large-T from lysates of simian virus 40-transformed 3T3 cells. This serum, designated F5, does not recognize antigenic determinants on native or denatured large-T and precipitates large-T because the 53K phosphoprotein forms a stable complex with large-T. This complex sediments at 23S on sucrose density gradients, corresponding to a molecular weight of 600K to 1,000K, and appears to contain only 53K and large-T as major components. It is held together by noncovalent bonds and is located in the cell nucleus. All the 53K immunoprecipitated from cell lysates by F5 is present in the high-molecular-weight complex, but large-T can be separated into a complexed and a free form on sucrose density gradients. The complexed form of large-T is more readily phosphorylated than the free form. We have been unable to detect an association of large-T with comparable host cell proteins during productive infections with simian virus 40.

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Selected References

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  1. Carroll R. B., Goldfine S. M., Melero J. A. Antiserum to polyacrylamide gel-purified simian virus 40 T antigen. Virology. 1978 Jun 1;87(1):194–198. doi: 10.1016/0042-6822(78)90171-x. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Chang C., Pancake S. J., Luborsky S. W., Mora P. T. Detergent solubilization and partial purification of tumor specific surface and transplantation antigens from SV40-virus-transformed mouse cells. Int J Cancer. 1977 Feb 15;19(2):258–266. doi: 10.1002/ijc.2910190216. [DOI] [PubMed] [Google Scholar]
  4. Chang C., Simmons D. T., Martin M. A., Mora P. T. Identification and partial characterization of new antigens from simian virus 40-transformed mouse cells. J Virol. 1979 Aug;31(2):463–471. doi: 10.1128/jvi.31.2.463-471.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. 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]
  8. 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]
  9. 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]
  10. Lamb R. A., Mahy B. W., Choppin P. W. The synthesis of sendai virus polypeptides in infected cells. Virology. 1976 Jan;69(1):116–131. doi: 10.1016/0042-6822(76)90199-9. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Lane D. P., Robbins A. K. An immunochemical investigation of SV40 T antigens. 1. Production properties and specificity of rabbit antibody to purified simian virus 40 large-T antigen. Virology. 1978 Jun 1;87(1):182–193. doi: 10.1016/0042-6822(78)90170-8. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Paucha E., Mellor A., Harvey R., Smith A. E., Hewick R. M., Waterfield M. D. Large and small tumor antigens from simian virus 40 have identical amino termini mapping at 0.65 map units. Proc Natl Acad Sci U S A. 1978 May;75(5):2165–2169. doi: 10.1073/pnas.75.5.2165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Polisky B., Kates J. Viral-specific polypeptides associated with newly replicated vaccinia DNA. Virology. 1975 Jul;66(1):128–139. doi: 10.1016/0042-6822(75)90184-1. [DOI] [PubMed] [Google Scholar]
  17. Prives C., Gluzman Y., Winocour E. Cellular and cell-free synthesis of simian virus 40 T-antigens in permissive and transformed cells. J Virol. 1978 Feb;25(2):587–595. doi: 10.1128/jvi.25.2.587-595.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Schmidt-Ullrich R., Thompson W. S., Lin P. S., Wallach D. F. Simian virus 40-specific proteins in the membranes of simian virus 40-transformed hamster and mouse cells. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5069–5072. doi: 10.1073/pnas.74.11.5069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Simons K., Helenius A., Leonard K., Sarvas M., Gething M. J. Formation of protein micelles from amphiphilic membrane proteins. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5306–5310. doi: 10.1073/pnas.75.11.5306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smart J. E., Ito Y. Three species of polyoma virus tumor antigens share common peptides probably near the amino termini of the proteins. Cell. 1978 Dec;15(4):1427–1437. doi: 10.1016/0092-8674(78)90066-1. [DOI] [PubMed] [Google Scholar]
  22. Smith A. E., Smith R., Paucha E. Characterization of different tumor antigens present in cells transformed by simian virus 40. Cell. 1979 Oct;18(2):335–346. doi: 10.1016/0092-8674(79)90053-9. [DOI] [PubMed] [Google Scholar]
  23. 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]

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