title: Isolation and characterization of conditional lethal amber nonsense mutants of vesicular stomatitis virus.
abstract: We describe the isolation and characterization of conditional lethal amber nonsense mutants of vesicular stomatitis virus (VSV), Indiana serotype. The mutants were isolated from a chemically mutagenized stock of wild-type virus by their ability to grow on genetically engineered cells which express a Xenopus laevis amber suppressor tyrosine tRNA gene (su+ cells) but not on the non-suppressor parental cells (su- cells). Five mutations were assigned to complementation group I (the L gene) and one to complementation group V (the G gene) by complementation analysis using temperature-sensitive mutants representing each of the five VSV cistrons. Four of the group I mutants were observed to synthesize a novel polypeptide species in su+ cells. Immunoprecipitation and immunoblotting studies using monospecific antisera directed against the N and C termini of the VSV L protein showed that the novel polypeptide species contain N terminal- but not C terminal-specific sequences and can thus be considered to be truncated versions of the L protein. In addition a protein which again contained N terminal- but not C terminal-specific sequences could be identified for the fifth group I mutant. Revertants of four of the group I mutants were isolated on the su- cells. The revertants all synthesized normal L protein but not the putative truncated version.
authors.affiliation: MRC Virology Unit, University of Glasgow, U.K.
authors.forename: B T
authors.initials: BT
authors.lastname: White
authors.forename: D J
authors.initials: DJ
authors.lastname: McGeoch
mesh_terms: D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics; D005804:Genes, Lethal; D005816:Genetic Complementation Test; D007158:Immunologic Techniques; D009154:Mutation; D013489:Suppression, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins