Expression of the Respiratory Syncytial Virus 22K Protein on the Surface of Infected HeLa Cells Free

Abstract

SUMMARY

Immunofluorescent staining of unfixed respiratory syncytial virus-infected HeLa cells with monoclonal antibodies (MAbs) demonstrated that the 22K protein is expressed on the cell membrane along with the fusion (F) protein and large glycoprotein (G). All three proteins were detected in the cytoplasm at 17 h postinfection and in the case of the F and G proteins this coincided with their appearance on the cell surface. However, the 22K protein could not be detected on the surface until approximately 16 h after its detection in the cytoplasm, when cytopathic effect was extensive. No evidence for the surface expression of the phosphoprotein (P), matrix (M) or nucleocapsid (N) proteins was found. Trypsin treatment of infected cells prior to unfixed immunofluorescent staining and Western blot analysis indicated that, unlike the G protein, the quantity of 22K protein detected on the cell surface constituted only a small proportion of the total present in the cell. A comparison of the patterns of immunofluorescent staining produced by MAbs on acetone-fixed infected cells suggested that the N, P and 22K proteins, but not the M protein, may be associated with the same intracellular structures.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-68-4-1217
1987-04-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/68/4/JV0680041217.html?itemId=/content/journal/jgv/10.1099/0022-1317-68-4-1217&mimeType=html&fmt=ahah

References

  1. Burnette W. N. 1981; ‘Western blotting’: electrophoretic transfer of proteins from sodium dodecyl sulfatepolyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  2. Collins P. L. 1985; The envelope-associated 22K protein of human respiratory syncytial virus: nucleotide sequence of the mRNA and a related polytranscript. Journal of Virology 54:65–71
    [Google Scholar]
  3. Dubovi E. J. 1982; Analysis of proteins synthesized in respiratory syncytial virus infected cells. Journal of Virology 42:372–378
    [Google Scholar]
  4. Gardner P. S., Mcquillin J. 1980; . Rapid Virus Diagnosis: Application of Immunofluorescence, 2nd edn.. London: Butterworths;
    [Google Scholar]
  5. Huang Y. T., Collins P. L., Wertz G. W. 1984; Identification of a new envelope-associated protein of human respiratory syncytial virus. In Non-segmented Negative Strand Viruses, Paramyxoviruses and Rhabdoviruses pp 365–368 Bishop D. H. L., Compans R. W. Edited by New York: Academic Press;
    [Google Scholar]
  6. Huang Y. T., Collins P. L., Wertz G. W. 1985; Characterization of the 10 proteins of human respiratory syncytial virus. Identification of a fourth envelope-associated protein. Virus Research 2:157–173
    [Google Scholar]
  7. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  8. Peeples M., Levine S. 1979; Respiratory syncytial virus polypeptides: their location in the virion. Virology 95:137–145
    [Google Scholar]
  9. Routledge E. G., Mcquillln J., Samson A. C. R. 1985; The development of monoclonal antibodies to respiratory syncytial virus and their use in diagnosis by indirect immunofluorescence. Journal of Medical Virology 15:305–320
    [Google Scholar]
  10. Routledge E. G., Willcocks M. M., Samson A. C. R., Scott R., Toms G. L. 1986; Respiratory syncytial virus glycoprotein expression in human and simian cell lines. Journal of General Virology 67:2059–2064
    [Google Scholar]
  11. Routledge E. G., Willcocks M. M., Morgan L., Samson A. C. R., Scott R., Toms G. L. 1987; Heterogeneity of the respiratory syncytial virus 22K protein revealed by Western blotting with monoclonal antibodies. Journal of General Virology 68:1209–1215
    [Google Scholar]
  12. Samson A. C. R., Willcocks M. M., Routledge E. G., Morgan L. A., Toms G. L. 1986; A neutralizing monoclonal antibody to respiratory syncytial virus which binds to both F1 and F2 components of the fusion protein. Journal of General Virology 67:1479–1483
    [Google Scholar]
  13. Scott R., Landajury M. D., Gardner P. S., Owen J. T. T. 1976; Detection of antibody to respiratory syncytial virus by membrane fluorescence. Clinical and Experimental Immunology 26:78–85
    [Google Scholar]
  14. Wunner W. H., Pringle C. R. 1976; Respiratory syncytial virus proteins. Virology 73:228–243
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-68-4-1217
Loading
/content/journal/jgv/10.1099/0022-1317-68-4-1217
Loading

Data & Media loading...

Most cited Most Cited RSS feed