1887

Abstract

The interaction of bovine respiratory syncytial virus (BRSV) phosphoprotein (P) with nucleocapsid (N) and large polymerase (L) proteins was investigated using an intracellular BRSV–CAT minigenome replication system. Coimmunoprecipitation assays using P-specific antiserum revealed that the P protein can form complexes with N and L proteins. Deletion mutant analysis of the P protein was performed to identify the regions of P protein that interact with N and L proteins. The results indicate that two independent N-binding sites exist on the P protein: an internal region of 161–180 amino acids and a C-terminal region of 221–241 amino acids. The L-binding site was mapped to a region of P protein encompassing amino acids 121–160. The data suggest that N and L protein binding domains on the P protein do not overlap.

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2001-04-01
2024-10-11
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References

  1. Asenjo A., Villanueva N. 2000; Regulated but not constitutive human respiratory syncytial virus (HRSV) P protein phosphorylation is essential for oligomerization. FEBS Letters 467:279–284
    [Google Scholar]
  2. Barr J., Easton A. J. 1995; Characterization of the interaction between the nucleoprotein and phosphoprotein of pneumonia virus of mice. Virus Research 39:221–235
    [Google Scholar]
  3. Buchholz U. J., Finke S., Conzelmann K.-K. 1999; Generation of bovine respiratory syncytial virus (BRSV) from cDNA: BRSV NS2 is not essential for replication in tissue culture, and the human RSV leader region acts as a functional BRSV genome promoter. Journal of Virology 73:251–259
    [Google Scholar]
  4. Byrappa S., Gavin D. K., Gupta K. C. 1995; A highly efficient procedure for site-specific mutagenesis of full-length plasmids using Vent DNA polymerase.. Genome Research 5404–407
    [Google Scholar]
  5. 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]
  6. Chenik M., Schnell M., Conzelmann K. K., Blondel D. 1998; Mapping the interacting domains between the rabies virus polymerase and phosphoprotein. Journal of Virology 72:1925–1930
    [Google Scholar]
  7. Collins J. K., Teegarden R. M., Macvean D. W., Smith G. H., Frank G., Salman S. 1988; Prevalence and specificity of antibodies to bovine respiratory syncytial virus in sera from feedlot and range cattle. American Journal of Veterinary Research 49:1316–1319
    [Google Scholar]
  8. Collins P. L., Hill M. G., Cristina J., Grosfeld H. 1996; Transcription elongation factor of respiratory syncytial virus, a nonsegmented negative strand RNA virus. Proceedings of the National Academy of Sciences, USA 93:81–85
    [Google Scholar]
  9. Curran J., Boeck R., Kolakofsky D. 1991; The Sendai virus P gene expresses both an essential protein and an inhibitor of RNA synthesis by shuffling modules via mRNA editing. EMBO Journal 10:3079–3085
    [Google Scholar]
  10. Garcia-Barreno B., Delgado T., Malero J. 1996; Identification of protein regions involved in the interaction of human respiratory syncytial virus phosphoprotein and nucleoprotein: significance for nucleocapsid assembly and formation of cytoplasmic inclusion. Journal of Virology 70:801–808
    [Google Scholar]
  11. Horikami S. M., Curran J., Kolakofsky D., Moyer S. A. 1992; Complexes of Sendai virus NP–P and P–L proteins are required for defective interfering particle genome replication in vitro. Journal of Virology 66:4901–4908
    [Google Scholar]
  12. Khattar S. K., Yunus A. S., Collins P. L., Samal S. K. 2000; Mutational analysis of the bovine respiratory syncytial virus nucleocapsid protein using a minigenome system: mutations that affect encapsidation, RNA synthesis, and interaction with the phosphoprotein. Virology 270:215–228
    [Google Scholar]
  13. 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]
  14. Ling R., Davis P. J., Qingzhong Y., Wood C. M., Pringle C. R., Cavanagh D., Easton A. J. 1995; Sequence and in vitro expression of the phosphoprotein gene of avian pneumovirus. Virus Research 36:247–257
    [Google Scholar]
  15. Lupas A., Van Dyke M., Stock J. 1991; Predicting coiled coils from protein sequences. Science 252:1162–1164
    [Google Scholar]
  16. 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]
  17. Mallipeddi S. K., Lupiani B., Samal S. K. 1996; Mapping the domains on the phosphoprotein of bovine respiratory syncytial virus required for N–P interaction using a two-hybrid system. Journal of General Virology 77:1019–1023
    [Google Scholar]
  18. Ryan K. W., Portner A. 1990; Separate domains of Sendai virus P protein are required for binding to viral nucleocapsid. Virology 174:515–521
    [Google Scholar]
  19. 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]
  20. Slack M. S., Easton A. J. 1998; Characterization of the interaction of the human respiratory syncytial virus phosphoprotein and nucleocapsid protein using the two-hybrid system. Virus Research 55:167–176
    [Google Scholar]
  21. Smallwood S. K., Ryan K. W., Moyer S. A. 1994; Deletion analysis defines a carboxyl-proximal region of Sendai virus P protein that binds to the polymerase L protein. Virology 202:154–163
    [Google Scholar]
  22. Stott E. J., Taylor G. 1985; Respiratory syncytial virus: brief review. Archives of Virology 84:1–52
    [Google Scholar]
  23. Takacs A. M., Das T., Banerjee A. K. 1993; Mapping of interacting domains between 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]
  24. van der Poel W. H., Brand A., Kramps J. A., Van Oirshot J. T. 1994; Respiratory syncytial virus infections in human beings and cattle. An epidemiological review. Journal of Infection 29:215–228
    [Google Scholar]
  25. Yunus A. S., Collins P. L., Samal S. K. 1998; Sequence analysis of functional polymerase (L) gene of bovine respiratory syncytial virus: determination of minimal trans -acting requirements for RNA replication. Journal of General Virology 79:2231–2238
    [Google Scholar]
  26. Zhao H., Banerjee A. K. 1995; Interaction between the nucleocapsid protein and the phosphoprotein of human parainfluenza virus 3. Journal of Biological Chemistry 21:12485–12490
    [Google Scholar]
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