1887

Abstract

SUMMARY

Yields of greater than 10 p.f.u./ml at 28 or 37 °C of the alphavirus Sindbis and the flavivirus Kunjin were obtained in the (Singh) cell line, the latent periods being 4 to 6 and 10 to 12 h, respectively. Despite a high background of host protein synthesis, virtually all the virus-specified proteins of the flaviviruses Kunjin, Dengue-2 and Japanese encephalitis were labelled and resolved by slab gel electrophoresis of infected and uninfected cell proteins. In contrast, only one induced protein, of mol. wt. 30000, was identified in cells labelled during Sindbis virus infection. The envelope glycoprotein V3 of Kunjin virus was resolved as a double band in samples of infected cytoplasm labelled with H-glucosamine, similar to that of carbohydrate-labelled V3 in vertebrate (Vero) cells. Attempts to reduce host protein synthesis selectively during labelling periods were unsuccessful using either a hypertonic inhibition block or treatment with 0.1 µg actinomycin D per ml. The most efficient labelling of Kunjin virus-specified proteins was achieved at 37 °C in the presence of actinomycin D. The largest non-structural flavivirus protein NV5 migrated slightly faster than NV5 from infected vertebrate (Vero) cells. The small non-structural proteins NV1, NV1½ and NV2 from infected mosquito cells were successively trimmed during post translational periods exceeding 70 min, compared to much shorter periods reported previously for post translational modifications of these proteins in vertebrate cells.

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1979-04-01
2024-12-08
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References

  1. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  2. Boulton R. W., Westaway E. G. 1977; Togavirus RNA: reversible effect of urea on genomes and absence of subgenomic viral RNA in Kunjin virus-infected cells. Archives of Virology 55:201–208
    [Google Scholar]
  3. Buckley S. M. 1969; Susceptibility of the Aedes albopictus and Aedes aegypti cell lines to infection with arboviruses. Proceedings of the Society for Experimental and Biology and Medicine 131:625–630
    [Google Scholar]
  4. Davey M. W., Daloarno L. 1974; Semliki Forest virus replication in cultures A. albopictus cells: studies on the establishment of persistence. Journal of General Virology 24:453–463
    [Google Scholar]
  5. Doherty R. L., Carley J. G., Mackerras M. J., Marks E. M. 1963; Studies of arthropod-borne virus infections in Queensland. III. Isolation and characterization of virus strains from wild caught mosquitoes in Northern Queensland. Australian Journal of Experimental Biology and Medical Science 41:17–39
    [Google Scholar]
  6. Eaton B. T., Regnery R. L. 1975; Polysomal RNA in Semliki Forest virus infected Aedes albopictus cells. Journal of General Virology 29:35–49
    [Google Scholar]
  7. Igarashi A., Koo R., Stollar V. 1977; Evolution and properties of Aedes albopictus cell cultures persistently infected with Sindbis virus. Virology 82:69–83
    [Google Scholar]
  8. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  9. Nuss D. L., Oppermann H., Koch G. 1975; Selective blockage of initiation of host protein synthesis in RNA-virus infected cells. Proceedings of the National Academy of Sciences of the United States of America 72:1258–1262
    [Google Scholar]
  10. Riedel B., Brown D. T. 1977; Role of extracellular virus in the maintenance of the persistent infection induced in Aedes albopictus (mosquito) cells by Sindbis virus. Journal of Virology 23:554–561
    [Google Scholar]
  11. Saborio J. L., Pong S. S., Koch G. 1974; Selective and reversible inhibition of initiation of protein synthesis in mammalian cells. Journal of Molecular Biology 85:195–211
    [Google Scholar]
  12. Shenk T. E., Koshelnyk K. A., Stollar V. 1974; Temperature sensitive virus from Aedes albopictus cells chronically infected with Sindbis virus. Journal of Virology 13:439–447
    [Google Scholar]
  13. Stollar V., Stollar B. D., Koo R., Harrap K. A., Schlesinger R. W. 1976; Sialic acid contents of Sindbis virus from vertebrate and mosquito cells–equivalence of biological and immunological viral properties. Virology 69:104–115
    [Google Scholar]
  14. Strauss J. H., Burge B. W., Darnell J. E. 1969; Sindbis virus infection of chicks and hamster cells: synthesis of virus-specific proteins. Virology 37:367–376
    [Google Scholar]
  15. Studier F. W. 1973; Analysis of bacteriophage T7 early RNAs and proteins on slab gels. Journal of Molecular Biology 79:237–248
    [Google Scholar]
  16. Westaway E. G. 1966; Assessment and application of a cell line from pig kidney for plaque assay and neutralization tests with twelve group B arboviruses. American Journal of Epidemiology 84:439–456
    [Google Scholar]
  17. Westaway E. G. 1973; Proteins specified by group B togaviruses in mammalian cells during productive infections. Virology 51:445–465
    [Google Scholar]
  18. Westaway E. G. 1975; The proteins of Murray Valley encephalitis virus. Journal of General Virology 27:283–293
    [Google Scholar]
  19. Westaway E. G. 1977; Strategy of the flavivirus genome. Evidence of multiple internal initiation of translation of proteins specified by Kunjin virus in mammalian cells. Virology 80:320–335
    [Google Scholar]
  20. Westaway E. G., Mckimm J. L., Mcleod L. G. 1977; Heterogeneity among flavivirus proteins separated in slab gels. Archives of Virology 53:305–312
    [Google Scholar]
  21. Westaway E. G., Reedman B. M. 1969; Proteins of the group B arbovirus Kunjin. Journal of Virology 4:688–693
    [Google Scholar]
  22. Westaway E. G., Shew M. 1977; Proteins and glycoproteins specified by the flavivirus Kunjin. Virology 80:309–319
    [Google Scholar]
  23. Wright P. J., Bowden D. S., Westaway E. G. 1977; Unique peptide maps of the three largest proteins specified by the flavivirus Kunjin. Journal of Virology 24:651–661
    [Google Scholar]
  24. Wright P. J., Westaway E. G. 1977; Comparisons of the peptide maps of Kunjin virus proteins smaller than the envelope protein. Journal of Virology 24:662–672
    [Google Scholar]
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