title: Generation and characterization of influenza A viruses with altered polymerase fidelity.
abstract: Genetic diversity of influenza A viruses (IAV) acquired through the error-prone RNA-dependent RNA polymerase (RdRP) or through genetic reassortment enables perpetuation of IAV in humans through epidemics or pandemics. Here, to assess the biological significance of genetic diversity acquired through RdRP, we characterize an IAV fidelity variant derived from passaging a seasonal H3N2 virus in the presence of ribavirin, a purine analogue that increases guanosine-to-adenosine mutations. We demonstrate that a single PB1-V43I mutation increases selectivity to guanosine in A/Wuhan/359/95 (H3N2) and A/Vietnam/1203/04 (H5N1) viruses. The H5N1 PB1-V43I-recombinant virus replicates to comparable titres as the wild-type virus in vitro or in the mouse lungs. However, a decrease in viral population diversity at day 3 post inoculation is associated with a tenfold reduced lethality and neurotropism in mice. Applying a fidelity variant with reduced mutational frequency, we provide direct experimental evidence for the role of genetic diversity in IAV pathogenesis.
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Peter P H
authors.initials: PP
authors.lastname: Cheung
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Watson
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Ka-Tim
authors.initials: KT
authors.lastname: Choy
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Sin
authors.initials: S
authors.lastname: Fun Sia
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Diana D Y
authors.initials: DD
authors.lastname: Wong
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Leo L M
authors.initials: LL
authors.lastname: Poon
authors.affiliation: 1] Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK [2] Division of Infection and Immunity, Research Department of Infection, University College London, Gower Street, London WC1E 6BT, UK.
authors.forename: Paul
authors.initials: P
authors.lastname: Kellam
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Guan
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: J S
authors.initials: JS
authors.lastname: Malik Peiris
authors.affiliation: Li Ka Shing Faculty of Medicine, Centre of Influenza Research, School of Public Health, The University of Hong Kong, No. 21 Sassoon Road, Pokfulam, Hong Kong SAR, China.
authors.forename: Hui-Ling
authors.initials: HL
authors.lastname: Yen
mesh_terms: D000241:Adenosine / Q000235:genetics; D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D024882:Drug Resistance, Viral / Q000235:genetics; D005260:Female; D014644:Genetic Variation; D006151:Guanosine / Q000235:genetics; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics* / Q000472:pathogenicity; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000472:pathogenicity; D008168:Lung / Q000473:pathology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation / Q000187:drug effects*; D009976:Orthomyxoviridae Infections / Q000401:mortality / Q000473:pathology / Q000821:virology; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics* / Q000378:metabolism; D016865:Reassortant Viruses / Q000235:genetics* / Q000472:pathogenicity; D012254:Ribavirin / Q000494:pharmacology; D016019:Survival Analysis; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D056189:Viral Tropism
chemical_list: D000998:Antiviral Agents; D014764:Viral Proteins; D006151:Guanosine; D012254:Ribavirin; D012324:RNA-Dependent RNA Polymerase; D000241:Adenosine