DNA repair pathway in alkylated human cells: apurinic/apyrimidinic intermediate resolved by Escherichia coli endonuclease IV-coupled alkaline elution.
Apurinic/apyrimidinic (AP) sites were measured in HeLa cells by digestion of cellular DNA with Escherichia coli endonuclease IV, an AP-specific endonuclease, prior to alkaline elution. The absence of non-specific endonuclease activity allowed endonuclease IV-sensitive AP sites to be detected with the sensitivity of conventional alkaline elution. Cells that were alkylated with dimethyl sulfate contained AP sites that were repaired along with DNA single-strand breaks during a post-alkylation recovery period. These results show that DNA alkylation products are repaired, at least in part, via an AP intermediate suggesting a repair pathway initiated by DNA glycosylases followed by DNA incision by AP endonuclease.
D000478:Alkylation; D004260:DNA Repair; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D043223:Deoxyribonuclease IV (Phage T4-Induced); D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D013463:Sulfuric Acid Esters / Q000494:pharmacology
D029968:Escherichia coli Proteins; D013463:Sulfuric Acid Esters; D004706:Endodeoxyribonucleases; D043223:Deoxyribonuclease IV (Phage T4-Induced); C468972:endonuclease IV, E coli; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; C007482:dimethyl sulfate