Advertisement

Archives of Virology

, Volume 78, Issue 3–4, pp 177–190 | Cite as

Immunofluorescent sites in vero cells infected with the flavivirus Kunjin

  • Mah Lee Ng
  • J. S. Pedersen
  • Ban Hock Toh
  • E. G. Westaway
Original Papers

Summary

The sites of replication and of accumulation of viral macromolecules were examined using fluorescent antibodies to viral products and to cell organelles. Synthesis of envelope protein and its accumulation in a narrow rim around the nucleus were detected at 4 hours post infection; concurrently, a progressive change was observed in the rough endoplasmic reticulum from a fine to a coarse network emanating from around the nucleus. This change in the network was visible by light microscopy. The immunofluorescent sites of viral RNA synthesis, located by use of anti-double stranded RNA, extended from the perinuclear region in another fine network which included many small foci or vesicles; these sites were also visible by light microscopy late in infection. None of these changes were associated with any visible redistribution of actin, intermediate filaments or microtubules, and no nuclear involvement was detected. However, when microtubules were disrupted by vinblastine treatment of cells, the distribution of the immunofluorescent sites of viral RNA synthesis was modified and the virus yield was reduced by at least 10-fold. These results confirmed our biochemical studies showing separation of viral sites of RNA synthesis and translation, and the accumulation of envelope protein in nuclear-associated membranes. The relevance of these observations is discussed in relation to the reports of specific membrane structures induced in flavivirus-infected cells.

Keywords

Intermediate Filament Vero Cell Vinblastine Post Infection Rough Endoplasmic Reticulum 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Boulton, R. W., Westaway, E. G.: Replication of the flavivirus Kunjin: Proteins, glycoproteins, and maturation associated with cell membranes. Virology69, 416–430 (1976).Google Scholar
  2. 2.
    Boulton, R. W., Westaway, E. G.: Togavirus RNA: Reversible effect of urea on genomes and absence of subgenomic viral RNA in Kunjin virus-infected cells. Arch. Virol.55, 201–208 (1977).Google Scholar
  3. 3.
    Bretherton, L., Toh, B. H., Jack, I.: IgM autoantibody to intermediate filaments inMycoplasma pneumoniae infections. Clin. Immunol. Immunpathol.18, 425–430 (1981).Google Scholar
  4. 4.
    Calberg-Bacq, C.-M., Rentier-Delrue, F., Osterrieth, P. M., Duchesne, P. Y.: Electron microscopy studies on Banzi virus particle and its development in the suckling mice brains. J. Ultrastruct. Res.53, 193–203 (1975).Google Scholar
  5. 5.
    Cardiff, R. D., Russ, S. B., Brandt, W. E., Russell, P. K.: Cytological localization of dengue-2 antigens: An immunological study with ultrastructural correlation. Infect. Immun.7, 809–816 (1973).Google Scholar
  6. 6.
    Demsey A., Steere, R. L., Brandt, W. E., Veltri, B. J.: Morphology and development of dengue-2 virus employing the freeze-fracture and thinsection technique. J. Ultrastruct. Res.46, 103–116 (1974).Google Scholar
  7. 7.
    Filshie, B. K., Rehacek, J.: Studies of the morphology of Murray Valley encephalitis and Japanese encephalitis viruses growing in cultured mosquito cells. Virology34, 435–443 (1968).Google Scholar
  8. 8.
    Green, J., Griffiths, G., Louvard, D., Quinn, P., Warren, G.: Passage of viral membrane proteins through the Golgi complex. J. Mol. Biol.152, 663–698 (1981).Google Scholar
  9. 9.
    Kos, K. A., Shapiro, D., Vaitzus, Z., Russell, P. K.: Viral polypeptide composition of Japanese encephalitis virus-infected cell membranes. Arch. Virol.47, 217–224 (1975a).Google Scholar
  10. 10.
    Kos, K. A., Osborne, B. A., Goldsby, R. A.: Inhibition of group B arbovirus antigen production and replication in cells enucleated with cytochalasin B. J. Virology15, 913–917 (1975b).Google Scholar
  11. 11.
    Leary, K., Blair, C. D.: Sequential events in the morphogenesis of Japanese encephalitis virus. J. Ultrastruct. Res.72, 123–129 (1980).Google Scholar
  12. 12.
    Lewin, B.: Cell structure and genetics. In: Gene Expression2: Eucaryotic Chromosomes, 2nd ed., 11–70. New York: John Wiley 1980.Google Scholar
  13. 13.
    Lubiniecki, A. S., Henry, C. J.: Autoradiographic localization of RNA synthesis directed by arboviruses in the cytoplasm of infected BHK-21 cells. Proc. Soc. Exp. Biol. Med.145, 1165–1169 (1974).Google Scholar
  14. 14.
    Mathy, J. P., Baum, R., Toh, B. H.: Autoantibodies to ribosomes and Systemic lupus erythematosis. Clin. Exp. Immunol.41, 73–80 (1980).Google Scholar
  15. 15.
    Matsumura, T., Stollar, V., Schlesinger, R. W.: Studies on the nature of dengue viruses. V. Structure and development of dengue virus in Vero cells. Virology46, 344–355 (1971).Google Scholar
  16. 16.
    Murphy, F. A.: Togavirus morphology and morphogenesis. In:Schlesinger, R. W. (eds.), The Togaviruses: Biology, Structure, Replication, 241–316. New York: Academic Press 1980.Google Scholar
  17. 17.
    Plagemann, P. G.: Vinblastine sulphate: Metaphase arrest, inhibition of RNA synthesis, and cytotoxicity in Novikoff rat hepatoma cells. J. Nat. Cancer Inst.45, 589–595 (1970).Google Scholar
  18. 18.
    Shapiro, D., Kos, K., Brandt, W. E., Russell, P. K.: Membrane-bound proteins of Japanese encephalitis in virus-infected chick embryo cells. Virology48, 360–372 (1972).Google Scholar
  19. 19.
    Sriurairatna, S., Bhamapravati, N., Diwan, A. R., Halstead, S. B.: Ultrastructural studies on dengue virus infection of human lymphoblasts. Infect. Immun.20, 173–179 (1978).Google Scholar
  20. 20.
    Stohlman, S. A., Wisseman, C. L., Jr., Eylar, O. R., Silverman, D. J.: Dengue virus-induced modifications of host cell membranes. J. Virology16, 1017–1026 (1975).Google Scholar
  21. 21.
    Stollar, B. D., Stollar, V.: Immunofluorescent demonstration of double-stranded RNA in the cytoplasm of Sindbis virus-infected cells. Virology42, 276–280 (1970).Google Scholar
  22. 22.
    Takeda, H., Oya, A., Hashimoto, K., Yasuda, T., Yamada, M.-A.: Association of virus specific replicative ribonucleic acid with nuclear membrane in chick embryo cells infected with Japanese encephalitis virus. J. gen. Virol.38, 281–291 (1978).Google Scholar
  23. 23.
    Toh, B. H., Gallichio, H. A., Jeffrey, P. L., Livett, B. G., Muller, H. K., Cauchi, M. N., Clarke, F. M.: Anti-actin stains synapses. Nature (London)264, 648–650 (1976).Google Scholar
  24. 24.
    Watt, F. M., Harris, H.: Microtubule-organizing centres in mammalian cells in culture. J. Cell Science44, 103–121 (1980).Google Scholar
  25. 25.
    Westaway, E. G.: Proteins specified by group B togaviruses in mammalian cells during productive infections. Virology51, 454–465 (1973).Google Scholar
  26. 26.
    Westaway, E. G.: Replication of Flaviviruses. In:Schlesinger, R. W. (ed.), The Togaviruses: Biology, Structure, Replication, 531–581. New York: Academic Press 1980.Google Scholar
  27. 27.
    Westaway, E. G., Ng, M. L.: Replication of flaviviruses: Separation of translation sites of Kunjin virus proteins and of cell proteins. Virology106, 107–122 (1980).Google Scholar
  28. 28.
    Westaway, E. G., Shew, M., Della-Porta, A. J.: Reactions of purified hemagglutinating antigens of flaviviruses with 19 S and 7 S antibodies. Infect. Immun.11, 630–634 (1975).Google Scholar
  29. 29.
    Wright, P. J., Warr, H. M., Westaway, E. G.: Synthesis of glycoproteins in cells infected by the flavivirus Kunjin. Virology109, 418–427 (1981).Google Scholar

Copyright information

© Springer-Verlag 1983

Authors and Affiliations

  • Mah Lee Ng
    • 1
  • J. S. Pedersen
    • 1
  • Ban Hock Toh
    • 1
  • E. G. Westaway
    • 1
  1. 1.Department of Microbiology and of Pathology and ImmunologyMonash UniversityMelbourneAustralia

Personalised recommendations