Systematic Identification of H274Y Compensatory Mutations in Influenza A Virus Neuraminidase by High-Throughput Screening
Open Access
- 15 January 2013
- journal article
- research article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 87 (2), 1193-1199
- https://doi.org/10.1128/jvi.01658-12
Abstract
Compensatory mutations contribute to the appearance of the oseltamivir resistance substitution H274Y in the neuraminidase (NA) gene of H1N1 influenza viruses. Here, we describe a high-throughput screening method utilizing error-prone PCR and next-generation sequencing to comprehensively screen NA genes for H274Y compensatory mutations. We found four mutations that can either fully (R194G, E214D) or partially (L250P, F239Y) compensate for the fitness deficiency of the H274Y mutant. The compensatory effect of E214D is applicable in both seasonal influenza virus strain A/New Caledonia/20/1999 and 2009 pandemic swine influenza virus strain A/California/04/2009. The technique described here has the potential to profile a gene at the single-nucleotide level to comprehend the dynamics of mutation space and fitness and thus offers prediction power for emerging mutant species.This publication has 42 references indexed in Scilit:
- Hemagglutinin–neuraminidase balance confers respiratory-droplet transmissibility of the pandemic H1N1 influenza virus in ferretsProceedings of the National Academy of Sciences of the United States of America, 2011
- The essential genome of a bacteriumMolecular Systems Biology, 2011
- Pandemic H1N1 2009 influenza virus with the H275Y oseltamivir resistance neuraminidase mutation shows a small compromise in enzyme activity and viral fitnessJournal of Antimicrobial Chemotherapy, 2010
- High-resolution mapping of protein sequence-function relationshipsNature Methods, 2010
- Influenza Virus Evolution, Host Adaptation, and Pandemic FormationCell Host & Microbe, 2010
- Structure of the amantadine binding site of influenza M2 proton channels in lipid bilayersNature, 2010
- Conserved Herpesviral Kinase Promotes Viral Persistence by Inhibiting the IRF-3-Mediated Type I Interferon ResponseCell Host & Microbe, 2009
- Functional studies indicate amantadine binds to the pore of the influenza A virus M2 proton-selective ion channelProceedings of the National Academy of Sciences of the United States of America, 2008
- Production of infectious hepatitis C virus in tissue culture from a cloned viral genomeNature Medicine, 2005
- A DNA transfection system for generation of influenza A virus from eight plasmidsProceedings of the National Academy of Sciences of the United States of America, 2000