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Journal of General Virology header logo Journal of General Virology

Volume 65, Issue 6

Characterization of Two Recombination-Complementation Groups of Uukuniemi Virus Temperature-sensitive Mutants No Access

Abstract

SUMMARY

With the aim of isolating temperature-sensitive () mutants defective in virus maturation or glycoprotein transport, Uukuniemi virus, a bunyavirus, was mutagenized with -methyl-′-nitro--nitrosoguanidine. Out of 13 initial clones unable to grow at 39 °C (non-permissive temperature), five mutants which grew to titres above 10 p.f.u./ml at 32 °C (permissive temperature) were selected for further studies. The mutants fell into two coinciding recombination-complementation groups. Three group I mutants (7, 8 and 12) and two group II mutants (6 and 11) synthesized all three RNA segments and were able to form the corresponding nucleoproteins at 39 °C. Thus, members of these two recombination groups had a RNA-positive phenotype. All five mutants showed immunofluorescence when cells were stained at 39 °C using a double-staining technique employing monoclonal antibodies against the glycoproteins G1 or G2, and polyclonal antibodies against the nucleoprotein, N. We have previously shown that in cells infected with wild-type virus both the G1/G2 and the N proteins accumulate in the Golgi complex, the site of virus maturation. In cells infected with 12, accumulation of G1 and G2, but not N protein, was observed in the Golgi complex at 39 °C. The N protein was found evenly scattered in the cytoplasm, suggesting lack of interaction between the G1/G2 and N proteins. With 6 and 11, G1 and G2 appeared to accumulate and aggregate in the endoplasmic reticulum (ER) at 39 °C. The location of the N protein coincided with that of the aggregated glycoproteins, suggesting that the N protein interacted with G1/G2 already in the ER. Thus, these mutants may prove valuable tools in studying the mechanism of Uukuniemi virus maturation.

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Keyword(s): maturation , ts mutants and Unkuniemi virus
© Journal of General Virology 1984
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1984-06-01
2025-01-25
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References

  1. Bishop D. H. L. 1979; Genetic potential of bunyaviruses. Current Topics in Microbiology and Immunology 86:1–13
     [Google Scholar]
  2. Bishop D. H. L., Shope R. E. 1979; Bunyaviridae. In Comprehensive Virology vol 14: pp. 1–175 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
     [Google Scholar]
  3. Bishop D. H. L., Calisher C. H., Casals J., Chumakov M. P., Gaidamovich S. Y., Hannoun C., Lvov D. K., Marshall J. D., Oker-blom N., Pettersson R. F., Porterfield J. S., Russell P. K., Shope R. E., Westaway E. G. 1980; Bunyaviridae. Intervirology 14:125–143
     [Google Scholar]
  4. Burge B. W., Pfefferkorn E. R. 1966; Isolation and characterization of conditional-lethal mutants of Sindbis virus. Virology 30:204–213
     [Google Scholar]
  5. Cash P., Vezza A. C., Gentsch J. R., Bishop D. H. L. 1979; Genome complexities of the three mRNA species of snowshoe hare bunyavirus and in vitro translation of S mRNA to viral N polypeptide. Journal of Virology 31:685–694
     [Google Scholar]
  6. Fuller F., Bishop D. H. L. 1982; Identification of viral coded non-structural polypeptides in bunyavirus-infected cells. Journal of Virology 41:643–648
     [Google Scholar]
  7. Fuller F., Bhown A. S., Bishop D. H. L. 1983; Bunyavirus nucleoprotein, N, and a non-structural protein, NSs, are coded by overlapping reading frames in the S RNA. Journal of General Virology 64:1705–1714
     [Google Scholar]
  8. Gentsch J., Bishop D. H. L. 1976; Recombination and complementation between temperature-sensitive mutants of a bunyavirus, snowshoe hare virus. Journal of Virology 20:351–354
     [Google Scholar]
  9. Gentsch J. R., Bishop D. H. L. 1978; Small viral RNA segment of bunyaviruses codes for viral nucleocapsid protein. Journal of Virology 28:417–419
     [Google Scholar]
  10. Gentsch J. R., Bishop D. H. L. 1979; M viral RNA segment of bunyaviruses codes for two glycoproteins, G1 and G2. Journal of Virology 30:767–770
     [Google Scholar]
  11. Gentsch J., Wynne L. R., Clewley J. P., Shope R. E., Bishop D. H. L. 1977; Formation of recombinants between snowshoe hare and La Crosse bunyaviruses. Journal of Virology 24:893–902
     [Google Scholar]
  12. Hewlett M. J., Pettersson R. F., Baltimore D. 1977; Circular forms of Uukuniemi virion RNA: an electron microscopic study. Journal of Virology 21:1085–1093
     [Google Scholar]
  13. Iroegbu C. U., Pringle C. R. 1981; Genetic interactions among viruses of the Bunyamwera complex. Journal of Virology 37:383–394
     [Google Scholar]
  14. Kuismanen E., Hedman K., Saraste J., Pettersson R. F. 1982; Uukuniemi virus maturation: accumulation of virus particles and viral antigens in the Golgi complex. Molecular Cell Biology 2:1444–1458
     [Google Scholar]
  15. Kuismanen E., Bang B., Hurme M., Pettersson R. F. 1984; Uukuniemi virus maturation: an immunofluorescence microscopy study using monoclonal glycoprotein-specific antibodies. Journal of Virology (in press)
    [Google Scholar]
  16. Murphy F. A., Harrison A. K., Whitfield S. G. 1973; Bunyaviridae: morphologic and morphogenetic similarities of Bunyamwera serologic supergroup viruses and several other arthropod-borne viruses. Intervirology 1:297–316
     [Google Scholar]
  17. Ozden S., Hannoun C. 1978; Isolation and preliminary characterization of temperature-sensitive mutants of Lumbo virus. Virology 84:210–212
     [Google Scholar]
  18. Ozden S., Hannoun C. 1980; Biochemical and genetic characteristics of Germiston virus. Virology 103:232–234
     [Google Scholar]
  19. Pesonen M., Kuismanen E., Pettersson R. F. 1982; Monosaccharide sequence of protein-bound glycans of Uukuniemi virus. Journal of Virology 41:390–400
     [Google Scholar]
  20. Pettersson R. F., Kaariainen L. 1973; The ribonucleic acids of Uukuniemi virus, a non-cubical tick-borne arbovirus. Virology 56:608–619
     [Google Scholar]
  21. Pettersson R., Kaariainen L., Von bonsdorff C.-H., Oker-blom N. 1971; Structural components of Uukuniemi virus, a non-cubical tick-borne arbovirus. Virology 46:721–729
     [Google Scholar]
  22. Pettersson R. F., Hewlett M. J., Baltimore D., Coffin J. M. 1977; The genome of Uukuniemi virus consists of three unique RNA segments. Cell 11:51–64
     [Google Scholar]
  23. Pringle C. R., Iroegbu C. U. 1982; Mutant identifying a third recombination group in a bunyavirus. Journal of Virology 42:873–879
     [Google Scholar]
  24. Ulmanen I., Seppala P., Pettersson R. F. 1981; In vitro translation of Uukuniemi virus-specific RNAs: identification of a nonstructural protein and a precursor to the membrane glycoproteins. Journal of Virology 37:72–79
     [Google Scholar]
  25. Von bonsdorff C. H., Pettersson R. 1975; Surface structure of Uukuniemi virus. Journal of Virology 16:1296–1307
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-65-6-1079
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Characterization of Two Recombination-Complementation Groups of Uukuniemi Virus Temperature-sensitive Mutants
J. Gen. Virol. 65, 1079 (1984); https://doi.org/10.1099/0022-1317-65-6-1079
/content/journal/jgv/10.1099/0022-1317-65-6-1079
/content/journal/jgv/10.1099/0022-1317-65-6-1079
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