Skip to main content
Log in

Localization of reverse-transcriptase in interferon-treated mouse cells chronically infected with Moloney leukemia virus

  • Original Papers
  • Published:
Archives of Virology Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Summary

Interferon treatment of mouse cells chronically infected with Moloney leukemia virus (3T3/MLV) resulted in 97 per cent inhibition of infective virus release.

The intracellular localization and distribution of virus reverse-transcriptase and group specific (gs) antigen were determined in interferon treated and control cells. Cytoplasm of infected cells was fractionated by isopycnic centrifugation on discontinuous sucrose gradients. Fractions were analysed for their chemical composition and characterized by the activity of membranal marker enzymes. The association and levels of viral antigens were determined in each fraction. Fractions enriched with 5′ nucleotidase, specific enzyme marker for plasma membrane, were also enriched with viral proteins. In interferon treated cells, intracellular accumulation of viral proteins was specifically localized in the plasma membrane. Threefold increase in reverse-transcriptase level was the maximal accumulation found in purified plasma membranes. Intracellular enzyme levels in interferon treated cells were in accordance with the amount of cell associated infective virus particles. The small accumulation of viral proteins and infective virus particles was not sufficient to account for the great reduction in virus yield observed in the supernatants of the interferon treated cells. A possible role for interferon in modification of plasma membrane associated with virus assembly is postulated.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Aboud, M., Weiss, O., Salzberg, S.: Rapid quantitation of interferon with chronically oncornavirus-producing cells. Infect. Immun.13, 1626–1632 (1976).

    Google Scholar 

  2. Billiau, A., Sobis, H., de Somer, P.: Influence of interferon on virus particles formation in different oncornavirus carrier cell lines. Int. J. Cancer12, 646–653 (1973).

    Google Scholar 

  3. Billiau, A., Hermans, H., Allen, P. T., Baron, S., de Somer, P.: Interferon inhibits C-type virus at a posttranscriptional, pre-release step. Arch. Virol.57, 205–220 (1978).

    Google Scholar 

  4. Caliguri, L. A., Tamm, I.: The role of cytoplasmic membranes in poliovirus biosynthesis. Virology42, 100–111 (1970).

    Google Scholar 

  5. Chang, E. H., Friedman, R. M.: A large glycoprotein of Moloney leukemia virus derived from interferon-treated cells. Biochem. biophys. Res. Commun.77, 392–397 (1977).

    Google Scholar 

  6. Chang, E. H., Myers, M. W., Wong, P. K. Y., Friedman, R. M.: The inhibitory effect of interferon on a temperature-sensitive mutant of Moloney murine leukemia virus. Virology77, 625–636 (1977).

    Google Scholar 

  7. Chang, E. H., Grollman, E. F., Jay, F. T., Lee, G., Kohn, L. D., Friedman, R. M.: Membrane alternations following interferon treatment. In:Stinebring, W. R., Chapple, P. J. (eds.), Advances in Experimental Medicine and Biology, Human interferon, Vol. 110, 85–99. New York-London: Plenum Press 1978.

    Google Scholar 

  8. Dianzani, F., Baron, S.: Activation by interferon of the event leading to the antiviral state. In:Baron, S., Dianzani, F. (eds.), The interferon system; A current review to 1978, Vol. 35, 297–302. Galveston, Texas: University of Texas Medical Branch 1977.

    Google Scholar 

  9. Dittmer, J. C., Wells, M. A.: Quantitative and qualitative analysis of lipids and lipid components. In:Colowick, S. P., Kaplan, S. O. (eds.), Methods in Enzymology, Vol. XIV, 482–532. New York: Academic Press 1969.

    Google Scholar 

  10. Eisenman, R. N., Vogt, V. M.: The biosynthesis of oncovirus proteins. Biochim. biophys. Acta473, 187–239 (1978).

    Google Scholar 

  11. Friedman, R. M., Chang, E. H., Ramseur, M. J., Myers, M. W.: Interferon directed inhibition of chronic murine leukemia virus production in cell cultures: Lack of effect on intracellular viral markers. J. Virol.16, 509–574 (1975).

    Google Scholar 

  12. Heppel, L. A., Hilmoe, R. J.: Purification and properties of 5′ nucleotidase. J. biol. Chem.188, 665–676 (1951).

    Google Scholar 

  13. Holland, J. J., Kiehn, E. D.: Influenza virus effects on cell membrane proteins. Science167, 202–205 (1970).

    Google Scholar 

  14. Ichihashi, Y., Matsumoto, S., Dales, S.: Biogenesis of poxviruses: Role of A-type inclusions and host cell membranes in virus dissemination. Virology46, 507–532 (1971).

    Google Scholar 

  15. Jainchill, L. J., Aaronson, A. S., Todaro, J. G.: Murine sarcoma and leukemia viruses: Assay using clonal lines of contact-inhibited mouse cells. J. Virol.4, 549–553 (1969).

    Google Scholar 

  16. Klement, V., Rowe, W. P., Hartley, J. M., Wendell, E. P.: Mixed culture cytopathogenicity: A new test for growth of murine leukemia viruses in tissue culture. Proc. Natl. Acad. Sci. U.S.A.63, 753–758 (1969).

    Google Scholar 

  17. Lenette, E. H., Schmidt, N. J.: Complement fixation test. In:Lenette, E. H., Schmidt, W. J. (eds.), Diagnostic procedures for viral and rickettsial infections, 52–58. New York: American Public Health Association, Inc. 1969.

    Google Scholar 

  18. Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measure ment with the Folin phenol reagent. J. biol. Chem.193, 265–275 (1951).

    Google Scholar 

  19. Munro, H. N., Fleck, A.: Recent developments in the measurements of nucleic acid in biological material. Analyst91, 78–88 (1966).

    Google Scholar 

  20. Naso, R. B., Arcement, L. J., Arlinghaus, R. B.: Biosynthesis of Rauscher leukemia viral proteins. Cell4, 31–36 (1975).

    Google Scholar 

  21. Pitha, P. M., Rowe, W. P., Oxman, M. N.: Effect of interferon on exogenous, endogenous, and chronic murine leukemia virus infection. Virology70, 324–338 (1976).

    Google Scholar 

  22. Pitha, P. M., Wivel, N. A., Fernie, B. F., Harper, H. P.: Effect of interferon on murine leukemia virus infection. IV. Formation of non-infectious virus in chronically infected cells. J. gen. Virol.42, 467–480 (1979).

    Google Scholar 

  23. Quigley, J. P., Rifkin, D. B., Reich, E.: Phospholipid composition of Rous sarcoma virus, host cell membranes and other enveloped RNA viruses. Virology46, 106–116 (1971).

    Google Scholar 

  24. Spear, P. G., Keller, J. M., Roisman, B.: Proteins specified by Herpes simplex virus II. Viral glycoproteins associated with cellular membranes. J. gen. Virol.5, 123–131 (1970).

    Google Scholar 

  25. Sherman, L., Teitz, Y.: Effect of interferon on biosynthesis of Moloney leukemia virus structural proteins. (Submitted for publication.)

  26. Svoboda, J.: The tumorogenic action of Rous sarcoma in rats and the permanent production of Rous virus by the induced rat sarcoma XC. Folia Biol.7, 77–81 (1961).

    Google Scholar 

  27. Swanson, M. A.: Glucose-6-phosphatase from liver. In:Colowick, S. P., Kaplan, N. O. (eds.), Methods in Enzymology, Vol. II, 541–543. New York: Academic Press 1957.

    Google Scholar 

  28. Teitz, Y., Lenette, E. H., Oshiro, L. S., Cremer, N.: Release of C-type particles from normal rat thymus cultures and those infected with MLV. J. Nat. Cancer Inst.46, 11–28 (1971).

    Google Scholar 

  29. Teitz, Y.: RNA dependent DNA polymerase in C-type particles from normal thymus cultures. Nature New Biol.232, 250–252 (1971).

    Google Scholar 

  30. Teitz, Y.: RNA directed DNA polymerase in C-type particles from normal rat thymus cultures and Moloney leukemia virus. In:Kohn, A., Shatkay, A. (eds.), Advances in Experimental Medicine and Biology, Control of gene expressions, Vol. 44, 329–356. New York-London: Plenum Press 1974.

    Google Scholar 

  31. Traub, A., Kaufmann, E., Ginzburg-Teitz, Y.: Studies on nuclear ribosomes. I. Association of DPN-pyrophosphorylase with nuclear ribosomes in normal and neoplastic tissues. Exp. Cell Res.34, 371–383 (1964).

    Google Scholar 

  32. Traub, A., Teitz, Y., Kaufmann, E.: A temporal and structural relationship between viral RNA polymerase and nicotinamide adenine dinucleotide pyrophosphorylase in the cytoplasm of Krebs-2 ascites cells infected with encephalomyocarditis virus. J. mol. Biol.36, 371–385 (1968).

    Google Scholar 

  33. Wharton, D. C., Tzagloff, A.: Cytochrome oxidase from beef heart mitochondria. In:Colowick, S. P., Kaplan, N. O. (eds.), Methods in Enzymology, Vol. X, 245–250. New York: Academic Press 1967.

    Google Scholar 

  34. Witte, O. N., Tsukamoto-Adey, A., Weissman, I. L.: Cellular maturation of oncornavirus glycoproteins: Topological arrangement of precursor and product forms in cellular membranes. Virology76, 539–553 (1977).

    Google Scholar 

  35. Wong, P. K. Y., Yuen, P. H., Macleod, R., Chang, E. H., Myers, M. W., Friedman, R. M.: The effect of interferon onde novo infection of Moloney murine leukemia virus. Cell10, 245–252 (1977).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

With 3 Figures

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sherman, L., Eylan, E. & Teitz, Y. Localization of reverse-transcriptase in interferon-treated mouse cells chronically infected with Moloney leukemia virus. Archives of Virology 66, 143–155 (1980). https://doi.org/10.1007/BF01314982

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01314982

Keywords

Navigation