title: Single mutation at the amino acid position 627 of PB2 that leads to increased virulence of an H5N1 avian influenza virus during adaptation in mice can be compensated by multiple mutations at other sites of PB2.
abstract: To understand the adaptation of H5N1 influenza viruses to mammals, a non-pathogenic influenza H5N1 virus (HN021) in mice was passaged for 15 times in mammalian host. Animal experimental results indicated that the mouse-adapted (MA) variants became highly pathogenic in mice after the passages. Sequence analysis showed that there was one amino acid substitution in PB2 protein of MA mutants after first passage (MA1), three amino acid substitutions in PB2 protein of MA5 and one amino acid in M1 protein, seven amino acids in HA protein and seven amino acids in PB2 protein of MA15, respectively. Animal experiments and growth assays with reassortant viruses produced by reverse genetics showed that mutations in PB2 alone contributed to the increase in virulence of HN021 in mice. Polymerase activity assays showed that the mutations in PB2 enhanced ribonucleoprotein complex polymerase activity in mammalian cells. Interestingly, one reverse mutation (K627E) took place at the amino acid position 627 of PB2 during passages of MA5 to MA15, indicating that a lysine at position 627 of PB2 is not absolutely needed for virulence and adaptation in mice by H5N1 virus. Taken together, the results suggest that mutations at multiple sites of PB2 contributed to the virulence and adaptation in mice, and the E627K mutation of PB2 is not an indispensable determinant in PB2 for mammalian adaptation by H5N1 avian influenza virus.
authors.affiliation: State Key Laboratory of Virology and Modern Virology Research Center, College of Life Sciences, Wuhan University, Luojia Hill, Wuhan 430072, PR China.
authors.forename: Junwei
authors.initials: J
authors.lastname: Li
authors.forename: Musarat
authors.initials: M
authors.lastname: Ishaq
authors.forename: Mabiala
authors.initials: M
authors.lastname: Prudence
authors.forename: Xiao
authors.initials: X
authors.lastname: Xi
authors.forename: Tao
authors.initials: T
authors.lastname: Hu
authors.forename: Qingzhen
authors.initials: Q
authors.lastname: Liu
authors.forename: Deyin
authors.initials: D
authors.lastname: Guo
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics*; D000818:Animals; D004252:DNA Mutational Analysis; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics* / Q000472:pathogenicity*; D008168:Lung / Q000473:pathology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D020125:Mutation, Missense; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*; D016865:Reassortant Viruses / Q000235:genetics / Q000472:pathogenicity; D012692:Serial Passage; D013489:Suppression, Genetic; D014764:Viral Proteins / Q000235:genetics*; D014774:Virulence
chemical_list: C057027:PB2 protein, Influenzavirus A; D014764:Viral Proteins; D012324:RNA-Dependent RNA Polymerase