1887

Abstract

SUMMARY

We have characterized the physiological defect in two temperature-sensitive mutants of the WSN strain of influenza virus which possessed a lesion in the haemagglutinin (HA) gene. In mutant virus-infected cells at the non-permissive temperature, the precursor HA polypeptide containing predominantly mannose-rich carbohydrate chains was not converted to the mature, functional HA polypeptide. Immunofluorescence showed that the HA polypeptide did not appear on the cell surface but was confined largely to the Golgi apparatus. It was concluded that the major physiological defect of these mutants was a block in the transport of the HA polypeptide beyond the Golgi apparatus. The block could be reversed, however, by lowering the temperature to 34 °C, resulting in normal processing of the precursor polypeptide and emergence of infectious progeny virus within 30 min. The HA activity of the two mutants, but not wild-type virus, was rapidly inactivated at 51 °C. Most, but not all, revertants derived from these mutants had HA with the heat stability of wild-type virus, suggesting that the temperature sensitivity and the heat lability of HA were two pleiotropic manifestations of a single lesion in the HA gene.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-65-11-1889
1984-11-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/65/11/JV0650111889.html?itemId=/content/journal/jgv/10.1099/0022-1317-65-11-1889&mimeType=html&fmt=ahah

References

  1. Basak S., Compans R. W. 1983; Studies on the role of glycosylation in the functions and antigenic properties of influenza virus glycoproteins. Virology 128:77–91
    [Google Scholar]
  2. Chamberlain J. P. 1979; Fluorographic detection of radioactivity in polyacrylamide gels with the water-soluble fluor, sodium salicylate. Analytical Biochemistry 98:132–135
    [Google Scholar]
  3. Davis A. R., Bos T., Ueda M., Nayak B. P., Dowbenko D., Compans R. W. 1983; Immune response to human influenza virus hemagglutinin expressed in Escherichia coli. Gene 21:273–284
    [Google Scholar]
  4. Lohmeyer J., Klenk H.-D. 1979; A mutant of influenza virus with a temperature-sensitive defect in the posttranslational processing of the hemagglutinin. Virology 93:134–145
    [Google Scholar]
  5. Maizel J. V. Jr 1971; Polyacrylamide gel electrophoresis of viral protein. Methods in Virology 5:179–246
    [Google Scholar]
  6. Palese P. 1977; The genes of influenza virus. Cell 10:1–10
    [Google Scholar]
  7. Palese P. 1978; The hemagglutinin gene of influenza viruses. In Topics in Infectious Disease vol 3: The Influenza Virus Hemagglutinin pp 49–57 Edited by Laver W. G., Bachmayer H., Weil R. Wien & New York: Springer-Verlag;
    [Google Scholar]
  8. Person S., Kousoulas K. G., Knowles R. W., Read G. S., Holland T. C., Keller P. M., Warner S. C. 1982; Glycoprotein processing in mutants of HSV-1 that induce cell fusion. Virology 117:293–306
    [Google Scholar]
  9. Sugiura A., Tobita K., Kilbourne E. D. 1972; Isolation and preliminary characterization of temperature-sensitive mutants of influenza virus. Journal of Virology 10:639–647
    [Google Scholar]
  10. Sugiura A., Ueda M., Tobita K., Enomoto C. 1975; Further isolation and characterization of temperature-sensitive mutants of influenza virus. Virology 65:363–373
    [Google Scholar]
  11. Sveda M. M., Markoff L. J., Lai C-J. 1982; Cell surface expression of the influenza virus hemagglutinin requires the hydrophobic carboxy-terminal sequences. Cell 30:649–656
    [Google Scholar]
  12. Sveda M. M., Markoff L. J., Lai C-J. 1984; Influenza virus hemagglutinin containing an altered hydrophobic carboxy terminus accumulates intracellularly. Journal of Virology 49:223–228
    [Google Scholar]
  13. Trentino A. L., Maley F. 1974; Purification and properties of an endo-β-N- acetylglucosaminidase from Streptomyces griseus. Journal of Biological Chemistry 249:811–817
    [Google Scholar]
  14. Ueda M., Kilbourne E. D. 1976; Temperature-sensitive mutants of influenza virus: a mutation in the hemagglutinin gene. Virology 70:425–431
    [Google Scholar]
  15. Virtanen I., Ekblom P., Laurila P. 1980; Subcellular compartmentalization of saccharide moieties in cultured normal and malignant cells. Journal of Cell Biology 85:429–434
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-65-11-1889
Loading
/content/journal/jgv/10.1099/0022-1317-65-11-1889
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error