Vaccinia DNA ligase complements Saccharomyces cerevisiae cdc9, localizes in cytoplasmic factories and affects virulence and virus sensitivity to DNA damaging agents.
The functional compatibility of vaccinia virus DNA ligase with eukaryotic counterparts was demonstrated by its ability to complement Saccharomyces cerevisiae cdc9. The vaccinia DNA ligase is a 63 kDa protein expressed early during infection that is non-essential for virus DNA replication and recombination in cultured cells. This implies complementation by a mammalian DNA ligase, yet no obvious recruitment of host DNA ligase I from the nucleus to the cytoplasm was observed during infection. An antiserum raised against a peptide conserved in eukaryotic DNA ligases identified the virus enzyme in discrete cytoplasmic 'factories', the sites of virus DNA synthesis, demonstrating immunological cross-reactivity between host DNA ligase I and the vaccinia enzyme. DNA ligase was not detected in the factories of a mutant virus lacking the ligase gene. Despite this, no difference in growth between wild-type (WT) and mutant virus was detectable even in Bloom's syndrome cells which have reduced DNA ligase I activity. However, DNA ligase negative virus showed an increased sensitivity to UV or bleomycin in cultured cells, and the importance of DNA ligase for virus virulence in vivo was demonstrated by the attenuated phenotype of the deletion mutant in intranasally infected mice.
Kerr
S M
SM
Sir William Dunn School of Pathology, University of Oxford, UK.
Johnston
L H
LH
Odell
M
M
Duncan
S A
SA
Law
K M
KM
Smith
G L
GL
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002460:Cell Line; D003593:Cytoplasm / Q000201:enzymology; D004249:DNA Damage; D011088:DNA Ligases / Q000378:metabolism*; D004279:DNA, Viral; D004285:Dogs; D005260:Female; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D005800:Genes, Fungal; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009153:Mutagens; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014616:Vaccinia virus / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D014774:Virulence / Q000235:genetics
D004279:DNA, Viral; D009153:Mutagens; D011088:DNA Ligases