THE CYSTEINE RESIDUES OF THE M2 PROTEIN ARE NOT REQUIRED FOR INFLUENZA-A VIRUS-REPLICATION

The M2 protein of influenza A virus functions as an ion channel. It co ntains three cysteine residues: cysteines 17 and 19, which form disulf ide bonds in the ectodomain, and cysteine 50, which is acylated. To un derstand the role of these cysteine residues in virus replication, we used reverse genetics to create influenza viruses in which the individ ual cysteines were mutated and a virus in which all three cysteines we re changed to serine. The M2 cysteine mutants that lacked either of th e cysteine residues in the ectodomain and the mutant that lacked all t hree residues had appreciably lower amounts of M2 oligomers than did t he wild-type virus when examined by sodium dodecyl sulfate-polyacrylam ide 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 p rotein to form functional oligomers for virus replication and that its cysteine residues are dispensable for influenza virus replication in vitro and in vivo. (C) 1997 Academic Press.