title: Identification, characterization, and natural selection of mutations driving airborne transmission of A/H5N1 virus.
abstract: Recently, A/H5N1 influenza viruses were shown to acquire airborne transmissibility between ferrets upon targeted mutagenesis and virus passage. The critical genetic changes in airborne A/Indonesia/5/05 were not yet identified. Here, five substitutions proved to be sufficient to determine this airborne transmission phenotype. Substitutions in PB1 and PB2 collectively caused enhanced transcription and virus replication. One substitution increased HA thermostability and lowered the pH of membrane fusion. Two substitutions independently changed HA binding preference from α2,3-linked to α2,6-linked sialic acid receptors. The loss of a glycosylation site in HA enhanced overall binding to receptors. The acquired substitutions emerged early during ferret passage as minor variants and became dominant rapidly. Identification of substitutions that are essential for airborne transmission of avian influenza viruses between ferrets and their associated phenotypes advances our fundamental understanding of virus transmission and will increase the value of future surveillance programs and public health risk assessments.
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Martin
authors.initials: M
authors.lastname: Linster
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: van Boheemen
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Miranda
authors.initials: M
authors.lastname: de Graaf
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Eefje J A
authors.initials: EJA
authors.lastname: Schrauwen
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Pascal
authors.initials: P
authors.lastname: Lexmond
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Mänz
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Theo M
authors.initials: TM
authors.lastname: Bestebroer
authors.affiliation: Institute of Virology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Baumann
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Debby
authors.initials: D
authors.lastname: van Riel
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Guus F
authors.initials: GF
authors.lastname: Rimmelzwaan
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Albert D M E
authors.initials: ADME
authors.lastname: Osterhaus
authors.affiliation: Institute of Virology, Philipps-University, 35043 Marburg, Germany.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Matrosovich
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands. Electronic address: r.fouchier@erasmusmc.nl.
authors.forename: Ron A M
authors.initials: RAM
authors.lastname: Fouchier
authors.affiliation: Department of Viroscience, Postgraduate School of Molecular Medicine, Erasmus Medical Center, 3015GE Rotterdam, the Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: Herfst
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D005289:Ferrets; D016679:Genome, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000378:metabolism; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000502:physiology*; D007251:Influenza, Human / Q000635:transmission* / Q000821:virology*; D009154:Mutation; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011991:Receptors, Virus / Q000378:metabolism; D012641:Selection, Genetic
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011991:Receptors, Virus; C483468:hemagglutinin, human influenza A virus; D012324:RNA-Dependent RNA Polymerase
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