title: Entry of vaccinia virus and cell-cell fusion require a highly conserved cysteine-rich membrane protein encoded by the A16L gene.
abstract: The vaccinia virus A16L open reading frame encodes a 378-amino-acid protein with a predicted C-terminal transmembrane domain and 20 invariant cysteine residues that is conserved in all sequenced members of the poxvirus family. The A16 protein was expressed late in infection and incorporated into intracellular virus particles with the N-terminal segment of the protein exposed on the surface. The cysteine residues were disulfide bonded via the poxvirus cytoplasmic redox system. Unsuccessful attempts to isolate a mutant virus with the A16L gene deleted suggested that the protein is essential for replication. To study the role of the A16 protein, we made a recombinant vaccinia virus that has the Escherichia coli lac operator system regulating transcription of the A16L gene. In the absence of inducer, A16 synthesis was repressed and plaque size and virus yield were greatly reduced. Nevertheless, virus morphogenesis occurred and normal-looking intracellular and extracellular virus particles formed. Purified virions made in the presence and absence of inducer were indistinguishable, though the latter had 60- to 100-fold-lower specific infectivity. A16-deficient virions bound to cells, but their cores did not penetrate into the cytoplasm. Furthermore, A16-deficient virions were unable to induce low-pH-triggered syncytium formation. The phenotype of the inducible A16L mutant was similar to those of mutants in which synthesis of the A21, A28, H2, or L5 membrane protein was repressed, indicating that at least five conserved viral proteins are required for entry of poxviruses into cells as well as for cell-cell fusion.
authors.affiliation: Laboratory of Viral Diseases, National Institutes of Health, 4 Center Dr., MSC 0445, Bethesda, MD 20892-0445, USA.
authors.forename: Suany
authors.initials: S
authors.lastname: Ojeda
authors.forename: Tatiana G
authors.initials: TG
authors.lastname: Senkevich
authors.forename: Bernard
authors.initials: B
authors.lastname: Moss
mesh_terms: D000595:Amino Acid Sequence; D002459:Cell Fusion; D002460:Cell Line; D003545:Cysteine; D008561:Membrane Fusion / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D014616:Vaccinia virus / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication
chemical_list: C516397:A16L protein, vaccinia virus; D014760:Viral Fusion Proteins; D003545:Cysteine