Complementation and analysis of an NP mutant of influenza virus.
Cell lines were constructed so as to express the influenza A virus nucleoprotein (NP) at levels approximating 5% of the total NP made throughout virus infection. Two types of cell lines were analyzed. One cell line (NP-5) expresses only the NP while another cell line was constructed which expresses the three viral polymerase proteins in addition to the NP (3PNP-4). Both cell lines were able to complement the growth of an NP mutant, ts56, at the non-permissive temperature. The 3PNP-4 cell line, constructed by transfecting a cell line already expressing the three polymerase proteins, continued to be able to complement viral PB2 mutants. In addition, sequence analysis was performed on the NP gene segment of A/WSN/33 and ts56 viruses. This analysis revealed that the mutant phenotype exhibited by ts56 at non-permissive temperature is due to a single serine to asparagine change (at codon 332) within the protein.
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Department of Microbiology, Mount Sinai School of Medicine of CUNY, NY 10029.
Palese
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D000595:Amino Acid Sequence; D001483:Base Sequence; D002461:Cell Line, Transformed; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D005816:Genetic Complementation Test; D015151:Immunoblotting; D009980:Influenza A virus / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins / Q000096:biosynthesis / Q000235:genetics*; D010641:Phenotype; D010957:Plasmids; D016601:RNA-Binding Proteins; D014162:Transfection; D014758:Viral Core Proteins; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics*
D004279:DNA, Viral; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D009698:Nucleoproteins; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; D014764:Viral Proteins