DNA repair pathway in alkylated human cells: apurinic/apyrimidinic intermediate resolved by Escherichia coli endonuclease IV-coupled alkaline elution

Abstract

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.