1887

Abstract

Specific interactions between the nucleocapsid protein (N) and the phosphoprotein (P) of bovine respiratory syncytial virus (BRSV) have been investigated using a yeast-based two-hybrid system. Plasmids encoding the yeast GAL4 DNA binding domain fused with the N gene and GAL4 activation domain fused with the P gene were cotransfected into competent yeast cells. The ability of the N and P proteins to interact was measured by activation of the reporter gene by the GAL4 transactivation region. Results indicated that the N and P proteins interact very strongly . When interactions between N and various deletion mutants of the P protein were examined, an internal region (aa 132–168) and the highly acidic C-terminal region (aa 236–241) of the P protein were found to be essential for N-P interaction. In addition, the highly basic N-terminal region (amino acids 1–40) was found to be involved in N-P interaction to a lesser extent.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-77-5-1019
1996-05-01
2024-05-02
Loading full text...

Full text loading...

/deliver/fulltext/jgv/77/5/JV0770051019.html?itemId=/content/journal/jgv/10.1099/0022-1317-77-5-1019&mimeType=html&fmt=ahah

References

  1. Baker J. C., Ames T. R., Markham R. J. F. 1986a; Seroepizootic study of bovine respiratory syncytial virus in a dairy herd. American Journal of Veterinary Research 47:240–245
    [Google Scholar]
  2. Baker J. C., Ames T. R., Werdin R. E. 1986b; Seroepizootic study of bovine respiratory syncytial virus in a beef herd. American Journal of Veterinary Research 47:246–253
    [Google Scholar]
  3. Barik S. 1992; Transcription of human respiratory syncytial virus genome RNA in vitro: requirement of cellular factor (s). Journal of Virology 66:6813–6818
    [Google Scholar]
  4. Chenik M., Chebli K., Gaudin Y., Blondel D. 1994; In vivo interaction of rabies virus phosphoprotein (P) and nucleoprotein (N): existence of two N-binding sites on P protein. Journal of General Virology 75:2889–2896
    [Google Scholar]
  5. Fields S., Song O. 1989; A novel genetic system to detect protein-protein interactions. Nature 340:245–246
    [Google Scholar]
  6. Gao Y., Lenard J. 1995; Multimerization and transcriptional activation of the phosphoprotein (P) of vesicular stomatitis virus by casein kinase-II. EMBO Journal 14:1240–1247
    [Google Scholar]
  7. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59
    [Google Scholar]
  8. 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]
  9. Kim H. W., Arrobio J. O., Brandt C. D., Jeffries B. C., Pyles G., Reid J. L., Chanock R. M., Parrott R. H. 1973; Epidemiology of respiratory syncytial virus in Washington D. C. 1. Importance of the virus in different respiratory tract disease syndromes and temporal distribution of infection. American Journal of Epidemiology 98:216–225
    [Google Scholar]
  10. Stott E. J., Taylor G. 1985; Respiratory syncytial virus: brief review. Archives of Virology 84:1–52
    [Google Scholar]
  11. Lerch R. A., Stott E. J., Wertz G. W. 1989; Characterization of bovine respiratory syncytial virus proteins and mRNAs and generation of cDNA clones to the viral mRNAs. Journal of Virology 63:833–840
    [Google Scholar]
  12. Mallipeddi S. K., Samal S. K., Mohanty S. B. 1990; Analysis of polypeptides synthesized in bovine respiratory syncytial virus-infected cells. Archives of Virology 115:23–36
    [Google Scholar]
  13. Mazumder B., Barik S. 1994; Requirement of casein kinase II-mediated phosphorylation for the transcriptional activity of human respiratory syncytial viral phosphoprotein P: transdominant negative phenotype of phosphorylation-defective P mutants. Virology 205:104–111
    [Google Scholar]
  14. Mazumder B., Adhikary G., Barik S. 1994; Bacterial expression of human respiratory syncytial viral phosphoprotein P and identification of Ser237 as the site of phosphorylation by cellular casein kinase II. Virology 205:93–103
    [Google Scholar]
  15. Örvell C., Norrby E., Mufson M. A. 1987; Preparation and characterization of monoclonal antibodies directed against five structural components of human respiratory syncytial virus subgroup B. Journal of General Virology 68:3125–3135
    [Google Scholar]
  16. Peeples M., Levine S. 1979; Respiratory syncytial virus polypeptides: their location in the virion. Virology 95:137–145
    [Google Scholar]
  17. Samal S. K., Pastey M. K., McPhillips T. H., Mohanty S. B. 1993; Bovine respiratory syncytial virus nucleocapsid protein expressed in insect cells specifically interacts with the phosphoprotein and the M2 protein. Virology 193:470–473
    [Google Scholar]
  18. Sãnchez-Seco M. P., Navarro J., Martinez R., Villanueva N. 1995; C-terminal phosphorylation of human respiratory syncytial virus P protein occurs mainly at serine residue 232. Journal of General Virology 76:425–130
    [Google Scholar]
  19. Stec D. S., Hill M. G., Collins P. L. 1991; Sequence analysis of the polymerase L gene of human respiratory syncytial virus and predicted phylogeny of non segmented negative-strand viruses. Virology 183:273–287
    [Google Scholar]
  20. Takacs A. M., Banerjee A. K. 1995; Efficient interaction of the vesicular stomatitis virus P protein with the L protein or the N protein in cells expressing the recombinant protein. Virology 208:821–826
    [Google Scholar]
  21. Takacs A. M., Das T., Banerjee A. K. 1993; Mapping of interacting domains between the nucleocapsid protein and the phosphoprotein of vesicular stomatitis virus by using a two-hybrid system. Proceedings of the National Academy of Sciences, USA 90:10375–10379
    [Google Scholar]
  22. Taylor G., Stott E. J., Bew M., Fernie B. F., Cote P. J., Collins A. P., Hughes M., Jebbett J. 1984; Monoclonal antibodies protect against respiratory syncytial virus infection in mice. Immunology 52:137–142
    [Google Scholar]
  23. 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/10.1099/0022-1317-77-5-1019
Loading
/content/journal/jgv/10.1099/0022-1317-77-5-1019
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