title: Cloning a eukaryotic DNA glycosylase repair gene by the suppression of a DNA repair defect in Escherichia coli.
abstract: If eukaryotic genes could protect bacteria with defects in DNA repair, this effect could be exploited for the isolation of eukaryotic DNA repair genes. We have thus cloned a DNA repair gene from Saccharomyces cerevisiae that directs the synthesis of a DNA glycosylase that specifically releases 3-methyladenine from alkylated DNA and in so doing protects alkylation-sensitive Escherichia coli from killing by methylating agents. The cloned yeast gene was then used to generate a mutant strain of S. cerevisiae that carries a defect in the glycosylase gene and is extremely sensitive to DNA methylation. This approach may allow the isolation of a large number of eukaryotic DNA repair genes.
authors.affiliation: Harvard School of Public Health, Charles A. Dana Laboratory of Toxicology, Boston, MA 02115.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: B
authors.initials: B
authors.lastname: Derfler
authors.forename: A
authors.initials: A
authors.lastname: Maskati
authors.forename: L
authors.initials: L
authors.lastname: Samson
mesh_terms: D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods; D004249:DNA Damage; D045647:DNA Glycosylases; D004260:DNA Repair; D004351:Drug Resistance; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D005800:Genes, Fungal; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D009699:N-Glycosyl Hydrolases / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
chemical_list: D011994:Recombinant Proteins; D008741:Methyl Methanesulfonate; C021565:3-methyladenine-DNA glycosylase; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases