Variations of the primary structure and conformation of the NP virus protein of influenza A recognized by monoclonal antibodies


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Abstract

The antigenic structure of influenza A NP protein was studied by using a panel of 12 monoclonal antibodies (mAbs). A group of strains of influenza-A virus with the known NP amino acid sequence was analyzed by radioimmunoprecipitation with a subsequent analysis of the immune complexes by electrophoresis in polyacrylamide gel. The examined group of strains comprised pairs of closely related variants like A/USSR/90/77 and A/Brazil/1/ 78 as well as A/Puerto Rico/8/34 (Mount Sinai variant) and A Puerto Rico/8/34 Cambridge 1 variant). The results made it possible to identify position 353 as a part of antigenic site. The combined results of radioimmunoprecipitation and immunoblotting suggest that the mentioned site is recognized by mAb IVE8 as a linear epitope and by mAb as a confirmation epitope. Amino acids in positions 196 and/or 290 are involved in the formation of another antigenic site. The N-terminal part of NP, removed in the course of protein processing, is not involved in the differentiation of avian and human strains by mAb 315. None of the mAbs used in our study recognized the N-terminal part of NP as a linear epitope.

References

  1. Кричевец С. Г., Варич Н. Л., Руднева И. А., Каверин Н. В. Зависимость антигенных свойств белка NP вирусов гриппа А человека и птиц от штаммовых вариаций аминокислотной последовательности // Вопр. вирусол. - 1999. - № 4. -С. 158-161.
  2. Семенова Н. П., Прокудина Е. И., Ямникова С. С., Вихрев Н. Е. Радиоиммунологический анализ антигенной вариабельности нуклеопротеина вируса гриппа // Молекул, генетика. - 1987. - № 11. - С. 31-33.
  3. Albo С., Valencia A., Portela A. Identification of an RNA binding region within the N-terminal third of the influenza A virus nucleoprotein // J. Virol. - 1995. - Vol. 69. - P. 3799-3806.
  4. Altmuller A., Fitch W. M., Scholtissek С. Biological and genetic evolution of the nucleoprotein gene of human influenza A vims // J. Gen. Virol. - 1989. -Vol.70. - P. 2111-2119.
  5. Biswas S. K., Boutz P. L., Nayak D. P. Influenza virus nucleoprotein interacts with influenza vims polymerase proteins // J. Virol. - 1998. - Vol. 72. - P. 5493-5501.
  6. Elton D., Medcalf L., Bishop K., Digard P. Oligomerization of influenza virus nucleoprotein: identification of positive and negative elements // Virology. - 1999. - Vol. 260. - P. 190-200.
  7. Gorman О. Т., Bean W. J., Kawaoka Y., Webster R. G. Evolution of the nucleoprotein gene of influenza A virus // J. Virol. - 1990. - Vol. 64. - P. 1487-1497.
  8. Gorman О. Т., Bean W. J., Kawaoka Y. et al. Evolution of influenza A virus nucleoprotein genes: implications for the origins of H1NI human and classical swine viruses // Ibid. - 1991. - Vol. 65. - P. 3704-3714.
  9. Herlocher L. M., Bucher D., Webster R. G. Host range determination and functional mapping of the nucleoprotein and matrix genes of influenza viruses using monoclonal antibodies // Virus Res. - 1992. - Vol. 22. - P. 281-293.
  10. Lopez J. A., Guillen M., Sanchez-Fauquier A., Melero J. A. An antigen-binding assay to determine the specificity of monoclonal antibodies against influenza virus and mapping of epitopes // J. Virol. Meth. - 1986. - Vol. 13. - P. 255-264.
  11. Mandler J., Gorman О. Т., Ludwig S. Derivation of the nucleoproteins (NP) of influenza A viruses isolated from marine mammals // Virology. - 1990. - Vol. 176. - P. 255-261.
  12. Martin-Benito J., Area E., Ortega J. et al. Three-dimensional reconstruction of a recombinant influenza virus ribonucleoprotein particle // EMBO Reports. - 2001. - Vol. 2. - P. 313-317.
  13. Shu L. L., Bean W. J., Webster R. G. Analysis of the evolution and variation of the human influenza A virus nucleoprotein gene from 1933 to 1990 // J. Virol. - 1993. - Vol. 67. - P. 2723-2729.
  14. Sklyanskaya E. I., Varich N. L., Amvrosieva T. V., Kaverin N. V. Virions and intracellular nucleocapsids produced during mixed heterotypic influenza infection of MDCK cells // Ibid. - 1985. - Vol. 53. - P. 679-683.
  15. Stitz L., Schmitz C., Binder D. et al. Characterization and immunological properties of influenza A virus nucleoprotein (NP): cell-associated NP isolated from infected cells or viral NP expressed by vaccinia recombinant virus do not confer protection // J. Gen. Virol. - 1990. - Vol. 78. - P. 1169-1179.
  16. Van Wyke K. L., Hinshaw V. S., Bean W. J., Webster R. G. Antigenic variation of influenza A virus nucleoprotein detected with monoclonal antibodies // J. Virol. - 1980. - Vol. 35. - P. 24-30.
  17. Van Wyke К. L., Bean W. J., Webster R. J. Monoclonal antibodies to the influenza A virus nucleoprotein affecting RNA transcription // Ibid. - 1981. - Vol. 39. - P. 313-317.
  18. Vareckova E., Betakova Т., Mucha V. et al. Preparation of monoclonal antibodies for the diagnosis of influenza A infection using different protocols // J. Immunol. Meth. - 1995. - Vol. 180. - P. 107-116.
  19. Weber F., Kochs G., Gruber S., Haller O. A classical bipartite nuclear localization signal on Thogoto and influenza A virus nucleoproteins // Virology. - 1998. - Vol. 250. - P. 9-18.
  20. Zhirnov O. P., Bukrinskaya A. G. Two forms of influenza virus nucleoprotein in infected cells and virions // Ibid. - 1981. - Vol. 109. - P. 174-179.
  21. Zhirnov O. P. Solubilization of matrix protein Ml/M from virions occurs at different pH for orthomyxo- and paramyxoviruses // Ibid. - 1990. - Vol. 176. - P. 274-279.
  22. Zhirnov O. P., Konakova T. E., Garten W., Klenk H.-D. Caspase-dependent N-terminal cleavage of influenza virus nucleocapsid protein in infected cells // J. Virol. - 1999. - Vol. 73. - P. 10158-10163.

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