A serine-to-asparagine mutation at position 314 of H5N1 avian influenza virus NP is a temperature-sensitive mutation that interferes with nuclear localization of NP
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We have generated a temperature-sensitive (ts) mutant from a human isolate of the H5N1 avian influenza virus by classical adaptation in cell culture. After 20 passages at low temperature, the virus showed a ts phenotype. The ts mutant also showed an attenuated phenotype after nasal inoculation in mice. Using reverse genetics, we generated reassortants carrying individual genomic segments of the wild-type and mutant viruses in an A/Puerto Rico/8/34 background, and found that the nucleoprotein (NP) gene could confer the ts phenotype. This mutant NP contains a serine-to-asparagine mutation at position 314 (S314N). The mutant NP protein showed a defect in nuclear localization at high temperature in mammalian cells.
KeywordsInfluenza Virus MDCK Cell Avian Influenza Virus H5N1 Avian Influenza Virus S314N Mutation
This work was supported by Thailand Research Fund (TRG5180011). S.N. and B.C. were supported by the postgraduate scholarship and research assistant programs, respectively, of the Faculty of Medicine Siriraj Hospital, pHw2000 plasmid was kindly provided by R.G. Webster.
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