1887

Abstract

The epitopes recognized by 41 monoclonal antibodies directed against the nucleocapsid protein (NP) of human parainfluenza virus type 2 (hPIV-2) were mapped on the primary structure of the hPIV-2 NP protein by testing their reactivities with deletion mutants. By Western immunoblotting using these monoclonal antibodies, the analysis of deletion mutants of the hPIV-2 NP protein was performed to identify the region essential for NP–NP interaction and phosphoprotein (P)-binding sites on the NP protein. The results indicate that the N-terminal 294 aa of the NP protein are all required for NP–NP self-assembly, and that two C-terminal parts of the NP protein are essential for NP–P binding: one region, aa 295–402, is required for binding to the C-terminal part of the P protein and another region, aa 403–494, to the N-terminal part of the P protein.

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1999-08-01
2020-01-25
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References

  1. Bankamp, B., Horikami, S. M., Thompson, P. D., Huber, M., Billeter, M. & Moyer, S. A. ( 1996; ). Domains of the measles virus N protein required for binding to P protein and self-assembly. Virology 216, 272-277.[CrossRef]
    [Google Scholar]
  2. Buchholz, C. J., Spehner, D., Drillien, R., Neubert, W. J. & Homann, H. E. ( 1993; ). The conserved N-terminal region of Sendai virus nucleocapsid protein NP is required for nucleocapsid assembly. Journal of Virology 67, 5803-5812.
    [Google Scholar]
  3. Buchholz, C. J., Retzler, C., Homann, H. E. & Neubert, W. J. ( 1994; ). The carboxy-terminal domain of Sendai virus nucleocapsid protein is involved in complex formation between phosphoprotein and nucleocapsid-like particles. Virology 204, 770-776.[CrossRef]
    [Google Scholar]
  4. Curran, J., Homann, H., Buchholz, C., Rochat, S., Neubert, W. & Kolakofsky, D. ( 1993; ). The hypervariable C-terminal tail of the Sendai paramyxovirus nucleocapsid protein is required for template function but not for RNA encapsidation. Journal of Virology 67, 4358-4364.
    [Google Scholar]
  5. Garcia-Barreno, B., Delegado, T. & Melero, J. A. ( 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 inclusions. Journal of Virology 70, 801-808.
    [Google Scholar]
  6. Homann, H. E., Willenbrink, W., Buchholz, C. J. & Neubert, W. J. ( 1991; ). Sendai virus protein–protein interactions studied by a protein-blotting protein-overlay technique: mapping of domains on NP protein required for binding to P protein. Journal of Virology 65, 1304-1309.
    [Google Scholar]
  7. 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]
  8. Horikami, S. M., Smallwood, S. & Moyer, S. A. ( 1996; ). The Sendai virus V protein interacts with the NP protein to regulate viral genome RNA replication. Virology 222, 383-390.[CrossRef]
    [Google Scholar]
  9. Krishnamurthy, S. & Samal, S. K. ( 1998; ). Identification of regions of bovine respiratory syncytial virus N protein required for binding to P protein and self-assembly. Journal of General Virology 79, 1399-1403.
    [Google Scholar]
  10. Miyahara, K., Kitada, S., Yoshimoto, M., Matsumura, H., Kawano, M., Komada, H., Tsurudome, M., Kusagawa, S., Nishio, M. & Ito, Y. ( 1992; ). Molecular evolution of human paramyxoviruses: nucleotide sequence analyses of the human parainfluenza type 1 virus NP and M protein genes and construction of phylogenetic trees for all the human paramyxoviruses. Archives of Virology 124, 255-268.[CrossRef]
    [Google Scholar]
  11. Myers, T. M., Pieters, A. & Moyer, S. A. ( 1997; ). A highly conserved region of the Sendai virus nucleocapsid protein contributes to the NP–NP binding domain. Virology 229, 322-335.[CrossRef]
    [Google Scholar]
  12. Nishio, M., Tsurudome, M., Kawano, M., Watanabe, N., Ohgimoto, S., Ito, M., Komada, H. & Ito, Y. ( 1996; ). Interaction between nucleocapsid protein (NP) and phosphoprotein (P) of human parainfluenza virus type 2: one of the two NP binding sites on P is essential for granule formation. Journal of General Virology 77, 2457-2463.[CrossRef]
    [Google Scholar]
  13. Nishio, M., Tsurudome, M., Ito, M., Watanabe, N., Kawano, M., Komada, H. & Ito, Y. ( 1997; ). Human parainfluenza virus type 2 phosphoprotein: mapping of monoclonal antibody epitopes and location of the multimerization domain. Journal of General Virology 78, 1303-1308.
    [Google Scholar]
  14. Ohgimoto, S., Bando, H., Kawano, M., Okamoto, K., Kondo, K., Tsurudome, M., Nishio, M. & Ito, Y. ( 1990; ). Sequence analysis of P gene of human parainfluenza type 2 virus: P and cysteine-rich proteins are translated by two mRNAs that differ by two nontemplated G residues. Virology 177, 116-123.[CrossRef]
    [Google Scholar]
  15. Ryan, K. W., Portner, A. & Murti, K. G. ( 1993; ). Antibodies to paramyxovirus nucleoproteins define regions important for immunogenicity and nucleocapsid assembly. Virology 193, 376-384.[CrossRef]
    [Google Scholar]
  16. Spehner, D., Drillien, R. & Howley, P. M. ( 1997; ). The assembly of the measles virus nucleoprotein into nucleocapsid-like particles is modulated by the phosphoprotein. Virology 232, 260-268.[CrossRef]
    [Google Scholar]
  17. Takacs, A. M., Dos, T. & Banerjee, A. K. ( 1993; ). Mapping of the interacting domains between the nucleoprotein and the phosphoprotein of vesicular stomatitis virus using a two-hybrid system. Proceedings of the National Academy of Sciences, USA 90, 10375-10379.[CrossRef]
    [Google Scholar]
  18. Takebe, Y., Seiki, M., Fujisawa, J., Hoy, P., Yokoto, K., Arai, K., Yoshida, M. & Arai, N. ( 1988; ). SR promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of the human T-cell leukemia virus type 1 long terminal repeat. Molecular and Cellular Biology 8, 466-472.
    [Google Scholar]
  19. Tsurudome, M., Nishio, M., Komada, H., Bando, H. & Ito, Y. ( 1989; ). Extensive antigenic diversity among human parainfluenza type 2 virus isolates and immunological relationships among paramyxoviruses revealed by monoclonal antibodies. Virology 171, 38-48.[CrossRef]
    [Google Scholar]
  20. Watanabe, N., Kawano, M., Tsurudome, M., Kusagawa, S., Nishio, M., Komada, H., Shima, T. & Ito, Y. ( 1996a; ). Identification of the sequences responsible for nuclear targeting of the V protein of human parainfluenza virus type 2. Journal of General Virology 77, 327-338.[CrossRef]
    [Google Scholar]
  21. Watanabe, N., Kawano, M., Tsurudome, M., Nishio, M., Ito, M., Ohgimoto, S., Suga, S., Komada, H. & Ito, Y. ( 1996b; ). Binding of the V proteins to the nucleocapsid proteins of human parainfluenza type 2 virus. Medical Microbiology and Immunology 185, 89-94.[CrossRef]
    [Google Scholar]
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