Vaccinia DNA ligase complements Saccharomyces cerevisiae cdc9, localizes in cytoplasmic factories and affects virulence and virus sensitivity to DNA damaging agents. EMBO J 1991 Dec;10(13):4343-50
Date
12/01/1991Pubmed ID
1756739Pubmed Central ID
PMC453187DOI
10.1002/j.1460-2075.1991.tb05012.xScopus ID
2-s2.0-0026061401 (requires institutional sign-in at Scopus site) 59 CitationsAbstract
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.
Author List
Kerr SM, Johnston LH, Odell M, Duncan SA, Law KM, Smith GLMESH terms used to index this publication - Major topics in bold
AnimalsBase Sequence
Blotting, Southern
Blotting, Western
Cell Line
Cytoplasm
DNA Damage
DNA Ligases
DNA, Viral
Dogs
Female
Fluorescent Antibody Technique
Gene Expression
Genes, Fungal
Mice
Mice, Inbred BALB C
Molecular Sequence Data
Mutagens
Recombination, Genetic
Saccharomyces cerevisiae
Vaccinia virus
Virulence
