title: The cysteine residues of the M2 protein are not required for influenza A virus replication.
abstract: The M2 protein of influenza A virus functions as an ion channel. It contains three cysteine residues: cysteines 17 and 19, which form disulfide bonds in the ectodomain, and cysteine 50 which is acylated. To understand the role of these cysteine residues in virus replication, we used reverse genetics to create influenza viruses in which the individual cysteines were mutated and a virus in which all three cysteines were changed to serine. The M2 cysteine mutants that lacked either of the cysteine residues in the ectodomain and the mutant that lacked all three residues had appreciably lower amounts of M2 oligomers than did the wild-type virus when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. None of the mutants, however, were defective in replication, either in vitro or in ferrets and mice. These findings demonstrate that noncovalent interactions are sufficient for the M2 protein to form functional oligomers for virus replication and that its cysteine residues are dispensable for influenza virus replication in vitro and in vivo.
authors.affiliation: Department of Virology, Istituto Superiore di Sanita, Rome, Italy.
authors.forename: M R
authors.initials: MR
authors.lastname: Castrucci
authors.forename: M
authors.initials: M
authors.lastname: Hughes
authors.forename: L
authors.initials: L
authors.lastname: Calzoletti
authors.forename: I
authors.initials: I
authors.lastname: Donatelli
authors.forename: K
authors.initials: K
authors.lastname: Wells
authors.forename: A
authors.initials: A
authors.lastname: Takada
authors.forename: Y
authors.initials: Y
authors.lastname: Kawaoka
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D003545:Cysteine; D017931:DNA Primers; D003673:Defective Viruses / Q000502:physiology; D005289:Ferrets; D006736:Horses; D009980:Influenza A virus / Q000472:pathogenicity / Q000502:physiology*; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009976:Orthomyxoviridae Infections / Q000503:physiopathology* / Q000821:virology; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012694:Serine; D014763:Viral Matrix Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D014779:Virus Replication; D000086082:Viroporin Proteins
chemical_list: D003545:Cysteine; D017931:DNA Primers; D012694:Serine; D014763:Viral Matrix Proteins; C031540:M-protein, influenza virus; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins