Abstract
Phenotypically mixed virus yields, obtained by coinfection of MDCK cells with influenza A/WSN/33 and B/Lee/40 viruses, contained both A/WSN/33 and B/Lee/40 NP proteins, as revealed by polyacrylamide gel electrophoresis of the purified 14C-amino acids-labeled virus. Virions were lysed with 0.6 M KCl-Triton X-100 buffer, and nucleocapsids were immunoprecipitated with antibodies against NP protein of influenza A virus. Polyacrylamide gel electrophoresis of the immunoprecipitate revealed NP protein of A/WSN/33 but not of B/Lee/40 virus. However, in similar experiments with the lysates of doubly infected cells, the band of B/Lee/40 NP protein was revealed in the polyacrylamide gel electrophoresis patterns of the immunoprecipitates. In an attempt to analyze the RNA content of the immune complexes, we absorbed the lysates of doubly infected [3H]uridine-labeled cells with protein A-containing Staphylococcus aureus covered with antibodies against the NP protein of influenza A virus. RNA extracted from the immune complexes contained genomic RNA segments of both A/WSN/33 and B/Lee/40 viruses. In control samples containing an artificial mixture of cell lysates separately infected with each virus, the analysis revealed homologous components (i.e., A/WSN/33 NP protein or RNA segments) in the immune complexes. The results suggest the presence of phenotypically mixed nucleocapsids in the cells doubly infected with influenza A and B viruses; in the course of the virion formation, the nucleocapsids lacking the heterologous NP protein are selected.
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- Bean W. J., Jr, Sriram G., Webster R. G. Electrophoretic analysis of iodine-labeled influenza virus RNA segments. Anal Biochem. 1980 Feb;102(1):228–232. doi: 10.1016/0003-2697(80)90343-7. [DOI] [PubMed] [Google Scholar]
- Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
- GOTLIEB T., HIRST G. K. The experimental production of combination forms of virus. III. The formation of doubly antigenic particles from influenza A and B virus and a study of the ability of individual particles of X virus to yield two separate strains. J Exp Med. 1954 Apr 1;99(4):307–320. doi: 10.1084/jem.99.4.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaverin N. V., Varich N. L., Sklyanskaya E. I., Amvrosieva T. V., Petrik J., Vovk T. C. Studies on heterotypic interference between influenza A and B viruses: a differential inhibition of the synthesis of viral proteins and RNAs. J Gen Virol. 1983 Oct;64(Pt 10):2139–2146. doi: 10.1099/0022-1317-64-10-2139. [DOI] [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- Kolakofsky D., Boy de la Tour E., Delius H. Molecular weight determination of Sendai and Newcastle disease virus RNA. J Virol. 1974 Feb;13(2):261–268. doi: 10.1128/jvi.13.2.261-268.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Londo D. R., Davis A. R., Nayak D. P. Complete nucleotide sequence of the nucleoprotein gene of influenza B virus. J Virol. 1983 Sep;47(3):642–648. doi: 10.1128/jvi.47.3.642-648.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McSharry J. J., Compans R. W., Choppin P. W. Proteins of vesicular stomatitis virus and of phenotypically mixed vesicular stomatitis virus-simian virus 5 virions. J Virol. 1971 Nov;8(5):722–729. doi: 10.1128/jvi.8.5.722-729.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pons M. W. Studies on the replication of influenza virus RNA. Virology. 1972 Mar;47(3):823–832. doi: 10.1016/0042-6822(72)90574-0. [DOI] [PubMed] [Google Scholar]
- Tobita K., Ohori K. Heterotypic interference between influenza viruses A/Aichi/2/68 and B/Massachusetts/1/71. Acta Virol. 1979 May;23(3):263–266. [PubMed] [Google Scholar]
- Winter G., Fields S. The structure of the gene encoding the nucleoprotein of human influenza virus A/PR/8/34. Virology. 1981 Oct 30;114(2):423–428. doi: 10.1016/0042-6822(81)90223-3. [DOI] [PubMed] [Google Scholar]
- Závada J. Viral pseudotypes and phenotypic mixing. Arch Virol. 1976;50(1-2):1–15. doi: 10.1007/BF01317996. [DOI] [PubMed] [Google Scholar]