Amplification of the ribonucleotide reductase small subunit gene: analysis of novel joints and the mechanism of gene duplication in vaccinia virus.
Amplification of the M2 gene encoding the small subunit of ribonucleotide reductase (EC 1.17.4.1) was analyzed in a collection of vaccinia virus (VV) isolates selected for resistance to 5 mM hydroxyurea (HU). Most of the mutants harbored tandem direct repeat arrays of the M2 gene, but several had duplicated M2 as an inverted repeat by genomic rearrangements involving the chromosomal termini. Novel joints formed by direct repeats were mapped, amplified in vitro, and sequenced. The junctions were simple fusions between DNA downstream and upstream of the M2 gene. Lack of sequence homology at the breakpoints indicated that the initial genomic rearrangements leading to gene amplification were due to nonhomologous recombination events.
Slabaugh
M B
MB
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-6503.
Roseman
N A
NA
Mathews
C K
CK
D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics; D004352:Drug Resistance, Microbial; D005796:Genes; D005814:Genes, Viral; D006918:Hydroxyurea / Q000494:pharmacology; D008969:Molecular Sequence Data; D005810:Multigene Family; D011995:Recombination, Genetic; D012264:Ribonucleotide Reductases / Q000235:genetics*; D014616:Vaccinia virus / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D014764:Viral Proteins; D012264:Ribonucleotide Reductases; D006918:Hydroxyurea