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. 1977 Mar;21(3):1170–1186. doi: 10.1128/jvi.21.3.1170-1186.1977

Simian virus 40 tumor-specific proteins: subcellular distribution and metabolic stability in HeLa cells infected with nondefective adenovirus type 2-simian virus 40 hybrid viruses.

W Deppert, G Walter, H Linke
PMCID: PMC515658  PMID: 66327

Abstract

HeLa cells infected with adenovirus type 2 (Ad2)-simian virus 40 (SV40) hybrid viruses produce several SV40-specific proteins. These include the previously reported 28,000-dalton protein of Ad2+ND1, and 42,000- and 56,000-dalton proteins of Ad2+ND2, the 56,000-dalton protein of Ad2+ND4, and the 42,000-dalton protein of Ad2+ND5. In this report, we extend the list of SV40-specific proteins induced by Ad2+ND4 to include proteins of apparent molecular weights of 28,000 42,000, 60,000, 64,000, 72,000, 74,000, and a doublet of 95,000. Cell fractionation studies demonstrate that the SV40-specific proteins are detectable in the nuclear, cytoplasmic, and plasma membrane fractions. By pulse-chase and cell fractionation experiments, three classes of SV40-specific proteins can be distinguished with regard to metabolic stability: (i) unstable in the cytoplasmic but stable in the nuclear and plasma membrane fractions; (ii) stable in the nuclear, cytoplasmic, and plasma membrane fractions; and (iii) unstable in all subcellular fractions. Immunoprecipitation of infected cell extracts demonstrates that most of the above proteins share antigenic determinants with proteins expressed in hamsters bearing SV40-induced tumors. Only the 42,000-dalton protein of Ad2+ND5 is not immunoprecipitable.

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Selected References

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  1. Anderson C. W., Gesteland R. F. Pattern of protein synthesis in monkey cells infected by simian virus 40. J Virol. 1972 May;9(5):758–765. doi: 10.1128/jvi.9.5.758-765.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Crumpacker C. S., Henry P. H., Kakefuda T., Rowe W. P., Levin M. J., Lewis A. M., Jr Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. 3. Base composition, molecular weight, and conformation of the Ad2+ND genome. J Virol. 1971 Mar;7(3):352–358. doi: 10.1128/jvi.7.3.352-358.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Del Villano B. C., Defendi V. Characterization of the SV40 T antigen. Virology. 1973 Jan;51(1):34–46. doi: 10.1016/0042-6822(73)90363-2. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Goldberg A. L., Dice J. F. Intracellular protein degradation in mammalian and bacterial cells. Annu Rev Biochem. 1974;43(0):835–869. doi: 10.1146/annurev.bi.43.070174.004155. [DOI] [PubMed] [Google Scholar]
  7. Grodzicker T., Anderson C., Sharp P. A., Sambrook J. Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1. J Virol. 1974 Jun;13(6):1237–1244. doi: 10.1128/jvi.13.6.1237-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Helling R. B., Goodman H. M., Boyer H. W. Analysis of endonuclease R-EcoRI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J Virol. 1974 Nov;14(5):1235–1244. doi: 10.1128/jvi.14.5.1235-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henry P. H., Schnipper L. E., Samaha R. J., Crumpacker C. S., Lewis A. M., Jr, Levine A. S. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. VI. Characterization of the DNA from five nondefective hybrid viruses. J Virol. 1973 May;11(5):665–671. doi: 10.1128/jvi.11.5.665-671.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Johnson F. B., Ozer H. L., Hoggan M. D. Structural proteins of adenovirus-associated virus type 3. J Virol. 1971 Dec;8(6):860–863. doi: 10.1128/jvi.8.6.860-863.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelly T. J., Jr, Lewis A. M., Jr Use of nondefective adenovirus-simian virus 40 hybrids for mapping the simian virus 40 genome. J Virol. 1973 Sep;12(3):643–652. doi: 10.1128/jvi.12.3.643-652.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kiehn E. D. Protein metabolism in SV40-infected cells. Virology. 1973 Nov;56(1):313–333. doi: 10.1016/0042-6822(73)90309-7. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [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. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  16. Lebowitz P., Kelly T. J., Jr, Nathans D., Lee T. N., Lewis A. M., Jr A colinear map relating the simian virus 40 (SV40) DNA segments of six adenovirus-SV40 hybrids to the DNA fragments produced by restriction endonuclease cleavage of SV40 DNA. Proc Natl Acad Sci U S A. 1974 Feb;71(2):441–445. doi: 10.1073/pnas.71.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levin M. J., Crumpacker C. S., Lewis A. M., Jr, Oxman M. N., Henry P. H., Rowe W. P. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. II. Relationship of adenovirus 2 deoxyribonucleic acid and simian virus 40 deoxyribonucleic acid in the Ad2+ND genome. J Virol. 1971 Mar;7(3):343–351. doi: 10.1128/jvi.7.3.343-351.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lewis A. M., Jr, Levin M. J., Wiese W. H., Crumpacker C. S., Henry P. H. A nondefective (competent) adenovirus-SV40 hybrid isolated from the AD.2-SV40 hybrid population. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1128–1135. doi: 10.1073/pnas.63.4.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lewis A. M., Jr, Levine A. S., Crumpacker C. S., Levin M. J., Samaha R. J., Henry P. H. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. V. Isolation of additional hybrids which differ in their simian virus 40-specific biological properties. J Virol. 1973 May;11(5):655–664. doi: 10.1128/jvi.11.5.655-664.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lewis A. M., Jr, Rowe W. P. Studies of nondefective Adenovirus 2-Simian virus 40 hybrid viruses. 8. Association of Simian virus 40 transplantation antigen with a specific region of the early viral genome. J Virol. 1973 Oct;12(4):836–840. doi: 10.1128/jvi.12.4.836-840.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. López-Revilla R., Walter G. Polypeptide specific for cells with adenovirus 2-SV40 hybrid Ad2+ND1. Nat New Biol. 1973 Aug 8;244(136):165–167. doi: 10.1038/newbio244165a0. [DOI] [PubMed] [Google Scholar]
  23. Morrow J. F., Berg P. Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3365–3369. doi: 10.1073/pnas.69.11.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Morrow J. F., Berg P., Kelly T. J., Jr, Lewis A. M., Jr Mapping of simian virus 40 early functions on the viral chromosome. J Virol. 1973 Sep;12(3):653–658. doi: 10.1128/jvi.12.3.653-658.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rose J. A., Maizel J. V., Jr, Inman J. K., Shatkin A. J. Structural proteins of adenovirus-associated viruses. J Virol. 1971 Nov;8(5):766–770. doi: 10.1128/jvi.8.5.766-770.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tai H. T., Smith C. A., Sharp P. A., Vinograd J. Sequence heterogeneity in closed simian virus 40 deoxyribonucleic acid. J Virol. 1972 Feb;9(2):317–325. doi: 10.1128/jvi.9.2.317-325.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Walter G., Martin H. Simian virus 40-specific proteins in HeLa cells infected with nondefective adenovirus 2-simian virus 40 hybrid viruses. J Virol. 1975 Nov;16(5):1236–1247. doi: 10.1128/jvi.16.5.1236-1247.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Walter G., Roblin R., Dulbecco R. Protein synthesis in Simian virus 40-infected monkey cells. Proc Natl Acad Sci U S A. 1972 Apr;69(4):921–924. doi: 10.1073/pnas.69.4.921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Williams J. F. Enhancement of adenovirus plaque formation on HeLa cells by magnesium chloride. J Gen Virol. 1970 Dec;9(3):251–255. doi: 10.1099/0022-1317-9-3-251. [DOI] [PubMed] [Google Scholar]

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