Fine structure mapping of five temperature-sensitive mutants in the 22- and 147-kilodalton subunits of vaccinia virus DNA-dependent RNA polymerase.
We have mapped the temperature-sensitive (ts) lesions of three mutants, ts51, ts53, and ts65, and two other mutants, ts7 and ts20, to regions on the vaccinia virus genome that encode the 147- and 22-kilodalton subunits of the viral DNA-dependent RNA polymerase, respectively. Plasmid and bacteriophage clones from the HindIII J region and the region spanning the HindIII J-H junction were used in marker rescue experiments to map the mutations. Sequence analysis of the region encoding the 22-kilodalton subunit in the wild-type, ts7, and ts20 viruses revealed a single base change in the mutants compared with that in the wild-type virus. The identification of these RNA polymerase mutants provides us with tools to understand transcription and its regulation in vaccinia virus.
Thompson
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CL
Department of Biochemistry, State University of New York, Buffalo 14214.
Hooda-Dhingra
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U
Condit
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RC
D000595:Amino Acid Sequence; D001483:Base Sequence; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D005796:Genes; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009154:Mutation; D011994:Recombinant Proteins / Q000235:genetics; D013696:Temperature; D014616:Vaccinia virus / Q000201:enzymology / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
D011994:Recombinant Proteins; D014764:Viral Proteins; D012321:DNA-Directed RNA Polymerases