Deletion of the Saccharomyces cerevisiae gene RAD30 encoding an Escherichia coli DinB homolog confers UV radiation sensitivity and altered mutability.
The dinB gene of Escherichia coli is an SOS-inducible gene of unknown function. Its mode of regulation and the amino acid sequence similarity of the predicted DinB protein to the UmuC protein of E. coli both suggest a role in cellular responses to DNA damage and probably in error-prone repair. Proteins with sequence similarity to DinB have been predicted from genes cloned from various prokaryotic and eukaryotic organisms, including Caenorhabditis elegans. Here we present the phenotypic characterization of a haploid Saccharomyces cerevisiae strain deleted for the ORF YDR419W, encoding a yeast DinB homolog. The deletion mutant is viable but is moderately sensitive to killing following exposure to ultraviolet (UV) radiation. Hence, we have named the gene RAD30. Steady-state levels of RAD30 transcripts are increased following UV irradiation. UV-induced locus-specific reversion of an ochre allele (arg4-17) is reduced in the rad30 deletion mutant. However, enhanced mutability was observed following treatment with the alkylating agent methylmethanesulfonate (MMS). Spontaneous mutability was also slightly increased. We conclude that RAD30 encodes an accessory function involved in DNA repair and mutagenesis. We speculate that the relatively weak phenotype and the opposite effects on mutability as a function of the type of DNA damage involved may derive from a functional redundancy of yeast proteins which facilitate replicative bypass of non-coding DNA lesions.
Roush
A A
AA
Graduate Program in Biochemistry, Cellular and Developmental Biology, Emory University School of Medicine, Atlanta, GA 30322, USA.
Suarez
M
M
Friedberg
E C
EC
Radman
M
M
Siede
W
W
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004260:DNA Repair; D004259:DNA-Directed DNA Polymerase; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D005656:Fungal Proteins / Q000235:genetics*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011836:Radiation Tolerance; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000528:radiation effects; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014466:Ultraviolet Rays; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
D001426:Bacterial Proteins; D004259:DNA-Directed DNA Polymerase; D029968:Escherichia coli Proteins; D005656:Fungal Proteins; D000094769:DNA Polymerase iota; C094399:DinB protein, E coli; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases