1887

Abstract

SUMMARY

Proteins induced in Newcastle disease virus (NDV)-infected chick embryo fibroblasts (CEF) and proteins incorporated into virions grown were analysed by modified versions of a two-dimensional polyacrylamide gel electrophoresis system. The following previously described NDV-induced proteins were detected and resolved from host proteins: L (mol. wt. approx. 200K), HN (75K), F (66K), F (56K), NP (55K) and M (39K). Two additional polypeptides, NAP (nucleocapsid-associated protein, mol. wt. 56K) and a 36 K mol. wt. protein were induced in NDV-infected cells. NAP but not 36K was found in purified virions. Radioactive labelling studies with H-glucosamine and P-orthophosphate demonstrated that neither NAP nor 36K is glycosylated, but that both are phosphorylated.

Variation in the isoelectric point and apparent mol. wt. of NAP among different strains was seen and exactly reproduced in both CEF and baby hamster kidney (BHK) cells. The synthesis of NDV-induced proteins including NAP and 36K was unaffected by actinomycin D, whereas the synthesis of host cell proteins was drastically reduced. These data are evidence that NAP and 36K are virus coded. Peptide analysis indicated that NAP, NP and F are unrelated polypeptides. The demonstration that NAP is virus coded, together with its phosphorylation and association with the nucleocapsid, suggest that NAP may be the NDV analogue of the P protein of Sendai virus.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-50-1-155
1980-09-01
2022-01-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/50/1/JV0500010155.html?itemId=/content/journal/jgv/10.1099/0022-1317-50-1-155&mimeType=html&fmt=ahah

References

  1. Ames G. F., Nikaido K. 1976; Two dimensional gel electrophoresis of membrane proteins. Biochemistry 15:616–623
    [Google Scholar]
  2. Ball L. A., Collins P. L., Hightower L. E. 1978; Transcription, translation and mapping of the genes of Newcastle disease virus. In Negative Strand Viruses and the Host Cell pp 367–382 Edited by Mahy B. W. J., Barry R. D. London: Academic Press;
    [Google Scholar]
  3. 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]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. 1977; Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. Journal of Biological Chemistry 252:1102–1106
    [Google Scholar]
  5. Clinton G. M., Burge B. W., Huang A. S. 1978; Effects of phosphorylation and pH on the association of NS with vesicular stomatitis virus cores. Journal of Virology 27:340–346
    [Google Scholar]
  6. Colonno R. J., Stone H. O. 1976; Isolation of a transcriptive complex from Newcastle disease virions. Journal of Virology 19:1080–1089
    [Google Scholar]
  7. Hightower L. E., Bratt M. A. 1974; Protein synthesis in Newcastle disease virus-infected chicken embryo cells. Journal of Virology 13:788–800
    [Google Scholar]
  8. Hightower L. E., Smith M. D. 1978; Effects of canavanine on protein metabolism in Newcastle disease virus-infected and uninfected chicken embryo cells. In Negative Strand Viruses and the Host Cell pp 395–405 Edited by Mahy B. W. J., Barry R. D. London: Academic Press;
    [Google Scholar]
  9. Hightower L. E., Morrison T. G., Bratt M. A. 1975; Relationships among the polypeptides of Newcastle disease virus. Journal of Virology 16:1599–1607
    [Google Scholar]
  10. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  11. Lamb R. A., Choppin P. W. 1977; The synthesis of Sendai virus polypeptides in infected cells. III. Phosphorylation of polypeptides. Virology 81:382–397
    [Google Scholar]
  12. Lamb R. A., Choppin P. W. 1978; Determination by peptide mapping of the unique polypeptides in Sendai virions and infected cells. Virology 84:469–478
    [Google Scholar]
  13. Madansky C. H., Bratt M. A. 1978; Non-cytopathic mutants of Newcastle disease virus: comparison with naturally occurring avirulent strains. In Negative Strand Viruses and the Host Cell pp 709–720 Edited by Mahy B. W. J., Barry R. D. London: Academic Press;
    [Google Scholar]
  14. Nagai Y., Klenk H.D., Rott R. 1976; Proteolytic cleavage of the viral glycoproteins and its significance for the virulence of Newcastle disease virus. Virology 72:494–508
    [Google Scholar]
  15. O’farrell P. H. 1975; High resolution two-dimensional electrophoresis of proteins. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  16. O’farrell P. Z., Goodman H. M., O’farrell P. H. 1977; High resolution two dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
    [Google Scholar]
  17. Orvell C. 1978; Structural polypeptides of mumps virus. Journal of General Virology 41:527–540
    [Google Scholar]
  18. Peluso R. W., Lamb R. A., Choppin P. W. 1977; Polypeptide synthesis in Simian virus 5-infected cells. Journal of Virology 23:177–187
    [Google Scholar]
  19. Privalsky M. L., Penhoet E. E. 1978; Influenza virus proteins: identity, synthesis and modification analysed by two dimensional gel electrophoresis. Proceedings of the National Academy of Sciences of the United States of America 75:3625–3629
    [Google Scholar]
  20. Raghow R., Portner A., Hsu C. H., Clarke S. B., Kingsbury D. W. 1978; Charge heterogeneity in polypeptides of negative strand RNA viruses. Virology 90:214–225
    [Google Scholar]
  21. Scheid A., Choppin P. W. 1973; Isolation and purification of the envelope proteins of Newcastle disease virus. Journal of Virology n:263–271
    [Google Scholar]
  22. Tyrrell D. L. J., Norrby E. 1978; Structural polypeptides of measles virus. Journal of General Virology 39:219–230
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-50-1-155
Loading
/content/journal/jgv/10.1099/0022-1317-50-1-155
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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