Role of uracil-DNA glycosylase in the repair of deaminated cytosine residues of DNA in Escherichia coli.
Uracil-DNA glycosylase, which acts specifically on uracil-containing DNA, was purified 250-fold from an extract of Escherichia coli 1100. The enzyme releases free uracil from DNA, producing alkali-labile apyrimidinic sites in the DNA. The enzyme is active on both native and heat-denatured DNA of phage PBS1, which contains uracil in place of thymine. piX174 DNA which had been treated with bisulfite and then at alkaline pH was susceptible to the action of uracil-DNA glycosylase. Since DNA treated with bisulfite alone was less susceptible to the enzyme, it is likely that the enzyme recognizes deaminated cytosine, namely uracil, but not bisulfite adducts of uracil and cytosine in the treated DNA. DNA treated with nitrite or hydroxylamine was not attacked by the enzyme. Enzyme activity acting on bisulfite-treated DNA was absent from an extract of E. coli mutant BD10 (ung). The mutant exhibited higher sensitivity to bisulfite than did the wild-type strain and was unable to reactivate phage T1 pre-exposed to bisulfite and weak alkali.
D001665:Binding Sites; D003090:Coliphages / Q000187:drug effects; D003596:Cytosine / Q000378:metabolism*; D004260:DNA Repair; D004279:DNA, Viral / Q000378:metabolism; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D009154:Mutation; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D009691:Nucleic Acid Denaturation; D013447:Sulfites / Q000494:pharmacology
D004279:DNA, Viral; D003856:Deoxyuracil Nucleotides; D013447:Sulfites; D003596:Cytosine; D009699:N-Glycosyl Hydrolases