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.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Blotting, Northern
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Blotting, Southern
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Cloning, Molecular* / methods
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DNA Damage*
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DNA Glycosylases*
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DNA Repair*
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Drug Resistance
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Escherichia coli / enzymology
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Escherichia coli / genetics*
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Escherichia coli / growth & development
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Genes, Fungal*
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Methyl Methanesulfonate / pharmacology
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N-Glycosyl Hydrolases / genetics*
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N-Glycosyl Hydrolases / metabolism
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Recombinant Proteins / metabolism
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Restriction Mapping
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics*
Substances
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Recombinant Proteins
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Methyl Methanesulfonate
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3-methyladenine-DNA glycosylase
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DNA Glycosylases
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N-Glycosyl Hydrolases