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. 1980 Sep;35(3):629–636. doi: 10.1128/jvi.35.3.629-636.1980

Sites of synthesis of viral proteins in avian sarcoma virus-infected chicken cells.

A F Purchio, S Jovanovich, R L Erikson
PMCID: PMC288856  PMID: 6252329

Abstract

We determined the sites of synthesis of avian sarcoma virus-specific proteins in infected chicken cells by immunoprecipitation of the products synthesized in vitro by free and membrane-bound polyribosomes; 85% of Pr76, the precursor of the viral internal structural proteins (group-specific antigens), was synthesized on free polyribosomes, and 15% was synthesized on membrane-bound polyribosomes. Pr92, the lycosylated precursor of the viral glycoproteins (gp85 and gp35), was synthesized exclusively on membrane-bound polyribomes, which is consistent with its role as a membrane protein. When we investigated the site of synthesis of pp60src, the product of the avian sarcoma virus src gene, we found that 90% was synthesized on free polyribosomes, whereas 10% was detected on membrane-bound polyribosomes. The implications of these results with respect to the subcellular location of pp60src are discussed.

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Selected References

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  1. Brugge J. S., Erikson E., Erikson R. L. Antibody to virion structural proteins in mammals bearing avian sarcoma virus-induced tumors. Virology. 1978 Feb;84(2):429–433. doi: 10.1016/0042-6822(78)90259-3. [DOI] [PubMed] [Google Scholar]
  2. Brugge J. S., Erikson R. L. Identification of a transformation-specific antigen induced by an avian sarcoma virus. Nature. 1977 Sep 22;269(5626):346–348. doi: 10.1038/269346a0. [DOI] [PubMed] [Google Scholar]
  3. Brugge J. S., Steinbaugh P. J., Erikson R. L. Characterization of the avian sarcoma virus protein p60src. Virology. 1978 Nov;91(1):130–140. doi: 10.1016/0042-6822(78)90361-6. [DOI] [PubMed] [Google Scholar]
  4. Buchhagen D. L., Hanafusa H. Intracellular precursors to the major glycoprotein of avian oncoviruses in chicken embryo fibroblasts. J Virol. 1978 Mar;25(3):845–851. doi: 10.1128/jvi.25.3.845-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chamberlain J. P. Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Anal Biochem. 1979 Sep 15;98(1):132–135. doi: 10.1016/0003-2697(79)90716-4. [DOI] [PubMed] [Google Scholar]
  6. Collett M. S., Erikson R. L. Protein kinase activity associated with the avian sarcoma virus src gene product. Proc Natl Acad Sci U S A. 1978 Apr;75(4):2021–2024. doi: 10.1073/pnas.75.4.2021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Diggelmann H. Biosynthesis of an unglycosylated envelope glycoprotein of Rous sarcoma virus in the presence of tunicamycin. J Virol. 1979 Jun;30(3):799–804. doi: 10.1128/jvi.30.3.799-804.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eisenman R. N., Vogt V. M. The biosynthesis of oncovirus proteins. Biochim Biophys Acta. 1978 Apr 6;473(3-4):187–239. doi: 10.1016/0304-419x(78)90014-8. [DOI] [PubMed] [Google Scholar]
  9. England J. M., Bolognesi D. P., Dietzschold B., Halpern M. S. Evidence that a precursor glycoprotein is cleaved to yield the major glycoprotein of avian tumor virus. J Virol. 1977 Feb;21(2):810–814. doi: 10.1128/jvi.21.2.810-814.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Erikson E., Collett M. S., Erikson R. L. In vitro synthesis of a functional avian sarcoma virus transforming-gene product. Nature. 1978 Aug 31;274(5674):919–921. doi: 10.1038/274919a0. [DOI] [PubMed] [Google Scholar]
  11. Ganoza M. C., Williams C. A. In vitro synthesis of different categories of specific protein by membrane-bound and free ribosomes. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1370–1376. doi: 10.1073/pnas.63.4.1370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hayward W. S. Size and genetic content of viral RNAs in avian oncovirus-infected cells. J Virol. 1977 Oct;24(1):47–63. doi: 10.1128/jvi.24.1.47-63.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hicks S. J., Drysdale J. W., Munro H. N. Preferential synthesis of ferritin and albumin by different populations of liver polysomes. Science. 1969 May 2;164(3879):584–585. doi: 10.1126/science.164.3879.584. [DOI] [PubMed] [Google Scholar]
  14. Kamine J., Buchanan J. M. Processing of 60,000-dalton sarc gene protein synthesized by cell-free translation. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4399–4403. doi: 10.1073/pnas.75.9.4399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Klemenz R., Diggelmann H. Extracellular cleavage of the glycoprotein precursor of Rous sarcoma virus. J Virol. 1979 Jan;29(1):285–292. doi: 10.1128/jvi.29.1.285-292.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Krueger J. G., Wang E., Goldberg A. R. Evidence that the src gene product of Rous sarcoma virus is membrane associated. Virology. 1980 Feb;101(1):25–40. doi: 10.1016/0042-6822(80)90480-8. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Lee J. S., Varmus H. E., Bishop J. M. Virus-specific messenger RNAs in permissive cells infected by avian sarcoma virus. J Biol Chem. 1979 Aug 25;254(16):8015–8022. [PubMed] [Google Scholar]
  19. Lodish H. F. Biosynthesis of reticulocyte membrane proteins by membrane-free polyribosomes. Proc Natl Acad Sci U S A. 1973 May;70(5):1526–1530. doi: 10.1073/pnas.70.5.1526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lodish H. F., Small B. Membrane proteins synthesized by rabbit reticulocytes. J Cell Biol. 1975 Apr;65(1):51–64. doi: 10.1083/jcb.65.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Montelaro R. C., Bolognesi D. P. Structure and morphogenesis of type-C retroviruses. Adv Cancer Res. 1978;28:63–89. doi: 10.1016/s0065-230x(08)60646-6. [DOI] [PubMed] [Google Scholar]
  22. Morrison T. G. Site of synthesis of membrane and nonmembrane proteins of vesicular stomatitis virus. J Biol Chem. 1975 Sep 10;250(17):6955–6962. [PubMed] [Google Scholar]
  23. Oppermann H., Bishop J. M., Varmus H. E., Levintow L. A joint produce of the genes gag and pol of avian sarcoma virus: a possible precursor of reverse transcriptase. Cell. 1977 Dec;12(4):993–1005. doi: 10.1016/0092-8674(77)90164-7. [DOI] [PubMed] [Google Scholar]
  24. Paterson B. M., Marciani D. J., Papas T. S. Cell-free synthesis of the precursor polypeptide for avian myeloblastosis virus DNA polymerase. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4951–4954. doi: 10.1073/pnas.74.11.4951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pawson T., Martin G. S., Smith A. E. Cell-free translation of virion RNA from nondefective and transformation-defective Rous sarcoma viruses. J Virol. 1976 Sep;19(3):950–967. doi: 10.1128/jvi.19.3.950-967.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  27. Purchio A. F., Erikson E., Brugge J. S., Erikson R. L. Identification of a polypeptide encoded by the avian sarcoma virus src gene. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1567–1571. doi: 10.1073/pnas.75.3.1567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Purchio A. F., Erikson E., Erikson R. L. Translation of 35S and of subgenomic regions of avian sarcoma virus RNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4661–4665. doi: 10.1073/pnas.74.10.4661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Redman C. M. Biosynthesis of serum proteins and ferritin by free and attached ribosomes of rat liver. J Biol Chem. 1969 Aug 25;244(16):4308–4315. [PubMed] [Google Scholar]
  30. Rohrschneider L. R. Immunofluorescence on avian sarcoma virus-transformed cells: localization of the src gene product. Cell. 1979 Jan;16(1):11–24. doi: 10.1016/0092-8674(79)90183-1. [DOI] [PubMed] [Google Scholar]
  31. Stohrer R., Hunter E. Inhibition of Rous sarcoma virus replication by 2-deoxyglucose and tunicamycin: identification of an unglycosylated env gene product. J Virol. 1979 Nov;32(2):412–419. doi: 10.1128/jvi.32.2.412-419.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vogt V. M., Eisenman R., Diggelmann H. Generation of avian myeloblastosis virus structural proteins by proteolytic cleavage of a precursor polypeptide. J Mol Biol. 1975 Aug 15;96(3):471–493. doi: 10.1016/0022-2836(75)90174-6. [DOI] [PubMed] [Google Scholar]
  33. Wang L. H., Duesberg P. H., Kawai S., Hanafusa H. Location of envelope-specific and sarcoma-specific oligonucleotides on RNA of Schmidt-Ruppin Rous sarcoma virus. Proc Natl Acad Sci U S A. 1976 Feb;73(2):447–451. doi: 10.1073/pnas.73.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weiss S. R., Varmus H. E., Bishop J. M. The size and genetic composition of virus-specific RNAs in the cytoplasm of cells producing avian sarcoma-leukosis viruses. Cell. 1977 Dec;12(4):983–992. doi: 10.1016/0092-8674(77)90163-5. [DOI] [PubMed] [Google Scholar]
  35. Willingham M. C., Jay G., Pastan I. Localization of the ASV src gene product to the plasma membrane of transformed cells by electron microscopic immunocytochemistry. Cell. 1979 Sep;18(1):125–134. doi: 10.1016/0092-8674(79)90361-1. [DOI] [PubMed] [Google Scholar]
  36. von der Helm K., Duesberg P. H. Translation of Rous sarcoma virus RNA in a cell-free system from ascites Krebs II cells. Proc Natl Acad Sci U S A. 1975 Feb;72(2):614–618. doi: 10.1073/pnas.72.2.614. [DOI] [PMC free article] [PubMed] [Google Scholar]

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