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. 1978 May;26(2):411–419. doi: 10.1128/jvi.26.2.411-419.1978

Biosynthesis, Immunological Specificity, and Intracellular Distribution of the Simian Virus 40-Specific Protein Induced by the Nondefective Hybrid Ad2+ND1

Gilbert Jay 1, Francis T Jay 2, Robert M Friedman 2, Arthur S Levine 1
PMCID: PMC354078  PMID: 207895

Abstract

Ad2+ND1, a nondefective hybrid virus containing a segment of the early region of simian virus 40 (SV40) DNA covalently inserted into the human adenovirus 2 genome, enhances the growth of human adenoviruses in simian cells and induces the SV40 U antigen. This hybrid previously has been shown to code for a 28,000 (28K) molecular weight protein not present in wild-type adenovirus 2-infected cells. By radioimmunoprecipitation using sera from hamsters bearing SV40-specific tumors, we have established that the Ad2+ND1-induced 28K protein is SV40-specific. This Ad2+ND1-induced protein is synthesized as a 30K molecular weight precursor, which is detectable only when infected cells are pulse-labeled in the presence of the protease inhibitor tosylamino phenylethyl chloromethyl ketone. Upon fractionation of labeled cell extracts, about 80% of the 28K protein is found in the plasma membrane fraction, whereas the remaining 20% is associated with the outer nuclear membrane. This protein is not detectable either in the nucleus or in the cytoplasm. Blockage of proteolytic cleavage by tosylamino phenylethyl chloromethyl ketone did not alter the topographic distribution of this SV40-specific protein, although the amount of the precursor protein in the outer nuclear membrane increased fourfold while that in the plasma membrane was proportionately decreased. This result suggests that the 28K protein is transferred from the outer nuclear membrane to the plasma membrane after posttranslational cleavage of the 30K precursor polypeptide. These data offer further support to the proposal that the 28K protein contains the determinants for SV40 U antigen and is responsible for SV40 enhancement of adenovirus growth in simian cells.

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Selected References

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  1. Anderson C. W., Lewis J. B., Baum P. R., Gesteland R. F. Simian virus 40-specific polypeptides in AD2+ ND1- and Ad2+ ND4-infected cells. J Virol. 1976 May;18(2):685–692. doi: 10.1128/jvi.18.2.685-692.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Deppert W., Walter G., Linke H. 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. J Virol. 1977 Mar;21(3):1170–1186. doi: 10.1128/jvi.21.3.1170-1186.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Grodzicker T., Lewis J. B., Anderson C. W. Conditional lethal mutants of adenovirus type 2-simian virus 40 hybrids. II. Ad2+ND1 host-range mutants that synthesize fragments of the Ad2+ND1 30K protein. J Virol. 1976 Aug;19(2):559–571. doi: 10.1128/jvi.19.2.559-571.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Horwitz M. S., Scharff M. D., Maizel J. V., Jr Synthesis and assembly of adenovirus 2. I. Polypeptide synthesis, assembly of capsomeres, and morphogenesis of the virion. Virology. 1969 Dec;39(4):682–694. doi: 10.1016/0042-6822(69)90006-3. [DOI] [PubMed] [Google Scholar]
  6. Jay G., Jay F. T., Friedman R. M., Levine A. S. Simian virus 40-specific ribosome-binding proteins induced by a nondefective adenovirus 2-simian virus 40 hybrid. J Virol. 1977 Sep;23(3):692–699. doi: 10.1128/jvi.23.3.692-699.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  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. 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]
  10. 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]
  11. Mann K., Hunter T., Walter G., Linke H. Evidence for simian virus 40 (SV40) coding of SV40 T-antigen and the SV40-specific proteins in HeLa cells infected with nondefective adenovirus type 2-SV40 hybrid viruses. J Virol. 1977 Oct;24(1):151–169. doi: 10.1128/jvi.24.1.151-169.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Nakajima K., Oda K. The alteration of ribosomes for mRNA selection concerned with adenovirus growth in SV40-infected simian cells. Virology. 1975 Sep;67(1):85–93. doi: 10.1016/0042-6822(75)90406-7. [DOI] [PubMed] [Google Scholar]
  14. Nakajima K., Oda K. The alteration of ribosomes for mRNA selection concerned with adenovirus growth in SV40-infected simian cells. Virology. 1975 Sep;67(1):85–93. doi: 10.1016/0042-6822(75)90406-7. [DOI] [PubMed] [Google Scholar]

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