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Genetic evidence for interaction between polypeptide PB2 of influenza virus and a host cell component
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NUCLEAR LOCALIZATION OF NSl AND NS2 PROTEINS IN INFLUENZA A VIRUS INFECTED CELLS:IDENTIFICATION OF NUCLEAR SIGNAL SEQUENCES P, PALESE, D. GREENSPAN, S. NAKADA and M. KRYSTAL
Dept. of Microbiology, Mt Sinai School of Medicine of CUNY, New York, N.Y. 10029, U.S.A. The genes coding for the NSl and NS2 proteins of influenza A/PR/8/34 virus were expressed in E. coli and rabbit antibodies against the bacterially expressed proms were obtained. Use of these monospecific antisera in immunofluorescence assays revealed the nuclear localization of both the NSl and NS2 proteins in infected cells. Based on an analysis of NS genes from several influenza virus variants, the nucleophilic signal sequence of the NSl protein appears to reside in the amino terminal half of the polypeptide. Expression studies using SV40 vectors containing NSl genes with site-specific mutations are underway to precisely determine the amino acids associated with the nuclear signal in the NSl protein. Similar studies are planned to determine the presence of nucleophilic signals in the NS2 protein of influenza A viruses.
79 GENETICEVIDENCEFOR INTERACTION BETWEENPOLYPEPTIDEPB2 OF INFLUENZAVIRUS AND A HOST CELL COMPONENT CHARLES R. PENN, Division of Molecular Biology, Animal Virus Research Institute, Pirbrigbt, Surrey GU24 ONF, U.K. A temperature-sensitivemutant of A/FFV/Ros~ackj34, t&47, is known to have a mutation at nucleotide 635 on virion RNA segment 8 that results in an altered polypeptide NS2 and a shortened NSl.
This mutation results in the ts
phenotype of tsC47, but does not cause the same ts phenotype in AIFM/l/47, which has the same nucleotide sequence around position 635 of RNA segment 8. Charaeterisationof reassortant viruses obtained by mixed infection with wild type FPV/Rostock, tsC47 and A/FMl1/47 has shown that tsC47 contains at least one additional mutation in RNA segment 1 (which encodes PB2) that acts synergistically with RNA segment 8 of tsC47 and of hf~~l~47 to contribute to a ts phenotype, and also confers a restricted host range on viruses containing mutant or wild type RNA segment 8. These data provide further evidence for interaction between PB2 and NSlf2 and also between PB2 and an unidentified host cell component. 40
NUCLEAR LOCALIZATION OF NSl AND NS2 PROTEINS IN INFLUENZA A VIRUS INFECTED CELLS:IDENTIFICATION OF NUCLEAR SIGNAL SEQUENCES P, PALESE, D. GREENSPAN, S. NAKADA and M. KRYSTAL
Dept. of Microbiology, Mt Sinai School of Medicine of CUNY, New York, N.Y. 10029, U.S.A. The genes coding for the NSl and NS2 proteins of influenza A/PR/8/34 virus were expressed in E. coli and rabbit antibodies against the bacterially expressed proms were obtained. Use of these monospecific antisera in immunofluorescence assays revealed the nuclear localization of both the NSl and NS2 proteins in infected cells. Based on an analysis of NS genes from several influenza virus variants, the nucleophilic signal sequence of the NSl protein appears to reside in the amino terminal half of the polypeptide. Expression studies using SV40 vectors containing NSl genes with site-specific mutations are underway to precisely determine the amino acids associated with the nuclear signal in the NSl protein. Similar studies are planned to determine the presence of nucleophilic signals in the NS2 protein of influenza A viruses.
79 GENETICEVIDENCEFOR INTERACTION BETWEENPOLYPEPTIDEPB2 OF INFLUENZAVIRUS AND A HOST CELL COMPONENT CHARLES R. PENN, Division of Molecular Biology, Animal Virus Research Institute, Pirbrigbt, Surrey GU24 ONF, U.K. A temperature-sensitivemutant of A/FFV/Ros~ackj34, t&47, is known to have a mutation at nucleotide 635 on virion RNA segment 8 that results in an altered polypeptide NS2 and a shortened NSl.
This mutation results in the ts
phenotype of tsC47, but does not cause the same ts phenotype in AIFM/l/47, which has the same nucleotide sequence around position 635 of RNA segment 8. Charaeterisationof reassortant viruses obtained by mixed infection with wild type FPV/Rostock, tsC47 and A/FMl1/47 has shown that tsC47 contains at least one additional mutation in RNA segment 1 (which encodes PB2) that acts synergistically with RNA segment 8 of tsC47 and of hf~~l~47 to contribute to a ts phenotype, and also confers a restricted host range on viruses containing mutant or wild type RNA segment 8. These data provide further evidence for interaction between PB2 and NSlf2 and also between PB2 and an unidentified host cell component. 40