title: Cytoplasmic domain of influenza B virus BM2 protein plays critical roles in production of infectious virus.
abstract: Influenza B virus BM2 is a type III integral membrane protein that displays H(+) ion channel activity. Analysis of BM2 knockout mutants has suggested that this protein is a necessary component for the capture of M1-viral ribonucleoprotein (vRNP) complex at the plasma membrane and for incorporation of vRNP complex into the virion during the assembly process. BM2 comprises 109 amino acid residues and possesses a longer cytoplasmic domain than the other 3 integral membrane proteins (hemagglutinin, neuraminidase, and NB). To explore whether the cytoplasmic domain of BM2 is important for infectious virus production, a series of BM2 deletion mutants lacking three to nine amino acid residues at the carboxyl terminus, BM2Delta107-109, BM2Delta104-109, and BM2Delta101-109, was generated by reverse genetics. Intracellular transport and incorporation into virions were indistinguishable between truncated BM2 proteins and wild-type BM2. The BM2Delta107-109 mutant produced levels of infectious virus similar to those of wild-type virus and displayed a spherical shape. However, the BM2Delta104-109 and BM2Delta101-109 mutants produced viruses containing dramatically reduced vRNP complex, as with BM2 knockout mutants, and formed enlarged, irregularly shaped virions. Moreover, gradient separation of membranes indicated that membrane association of M1 from mutants was greatly affected by carboxyl-terminal truncations of BM2. Studies of alanine substitution mutants further suggested that amino acid sequences in the 98-109 region are variable while those in the 86-97 region are a prerequisite for innate BM2 function. These results indicate that the cytoplasmic domain of the BM2 protein is required for firm association of the M1 protein with lipid membranes, vRNP complex incorporation into virions, and virion morphology.
authors.affiliation: Laboratory of Influenza Viruses, Department of Virology 3, National Institute of Infectious Diseases, Gakuen 4-7-1 Musashi-Murayama, Tokyo 208-0011, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Imai
authors.forename: Kazunori
authors.initials: K
authors.lastname: Kawasaki
authors.forename: Takato
authors.initials: T
authors.lastname: Odagiri
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry; D006801:Humans; D009981:Influenza B virus / Q000235:genetics / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000235:genetics / Q000502:physiology; D017434:Protein Structure, Tertiary; D012261:Ribonucleoproteins / Q000032:analysis; D017384:Sequence Deletion; D014763:Viral Matrix Proteins / Q000032:analysis; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000737:chemistry / Q000648:ultrastructure; D019065:Virus Assembly / Q000235:genetics / Q000502:physiology*
chemical_list: C064964:BM2 polypeptide, influenza B virus; D050505:Mutant Proteins; D012261:Ribonucleoproteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins