Giuseppe
Emeritus
J Virol. 2009 Feb 18. [Epub ahead of print]
Mutational analysis of conserved amino acids in the influenza A virus nucleoprotein.
Li Z, Watanabe T, Hatta M, Watanabe S, Nanbo A, Ozawa M, Kakugawa S, Shimojima M, Yamada S, Neumann G, Kawaoka Y. - Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706, USA; Division of Virology, Department of Microbiology and Immunology, and International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan; ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan.
The nucleoprotein (NP), which has multiple functions during the virus life cycle, possesses regions that are highly conserved among influenza A, B and C viruses.
To better understand the roles of highly conserved NP amino acids in viral replication, we conducted a comprehensive mutational analysis. Using reverse genetics, we attempted to generate 74 viruses possessing mutations at conserved amino acids of NP.
Of those, 48 mutant viruses were successfully rescued; 26 mutants were not viable, suggesting a critical role of the respective NP amino acids in viral replication. To identify the step(s) in the viral life cycle which is impaired by these NP mutations, we examined viral genome replication/transcription, NP localization, and incorporation of viral RNA segments into progeny virions.
We identified 15 amino acid substitutions in NP that inhibited viral genome replication and/or transcription, resulting in significant growth defect of viruses possessing these substitutions. We also found several NP mutations that affected the efficient incorporation of multiple viral RNA segments into progeny virions even though single vRNA segment was incorporated efficiently.
The respective conserved amino acids in NP may thus be critical for the assembly and/or incorporation of sets of eight vRNA segments.
PMID: 19225007 [PubMed - as supplied by publisher]
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Mutational analysis of conserved amino acids in the influenza A virus nucleoprotein.
Li Z, Watanabe T, Hatta M, Watanabe S, Nanbo A, Ozawa M, Kakugawa S, Shimojima M, Yamada S, Neumann G, Kawaoka Y. - Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, 2015 Linden Drive, Madison, WI 53706, USA; Division of Virology, Department of Microbiology and Immunology, and International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan; ERATO Infection-Induced Host Responses Project, Japan Science and Technology Agency, Saitama 332-0012, Japan.
The nucleoprotein (NP), which has multiple functions during the virus life cycle, possesses regions that are highly conserved among influenza A, B and C viruses.
To better understand the roles of highly conserved NP amino acids in viral replication, we conducted a comprehensive mutational analysis. Using reverse genetics, we attempted to generate 74 viruses possessing mutations at conserved amino acids of NP.
Of those, 48 mutant viruses were successfully rescued; 26 mutants were not viable, suggesting a critical role of the respective NP amino acids in viral replication. To identify the step(s) in the viral life cycle which is impaired by these NP mutations, we examined viral genome replication/transcription, NP localization, and incorporation of viral RNA segments into progeny virions.
We identified 15 amino acid substitutions in NP that inhibited viral genome replication and/or transcription, resulting in significant growth defect of viruses possessing these substitutions. We also found several NP mutations that affected the efficient incorporation of multiple viral RNA segments into progeny virions even though single vRNA segment was incorporated efficiently.
The respective conserved amino acids in NP may thus be critical for the assembly and/or incorporation of sets of eight vRNA segments.
PMID: 19225007 [PubMed - as supplied by publisher]
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