Repair of O6-ethylguanine in DNA by a chromatin fraction from rat liver: transfer of the ethyl group to an acceptor protein

Abstract

Incubation of O6-[3H]ethylguanine-containing DNA with a rat liver chromatin fraction resulted in a decrease in the O6-ethylguanine content of the DNA. Analysis of the products of this reaction showed that the ethyl group had been transferred from the O6-ethylguanine to a protein acceptor. When the incubation mixture was separated on a cesium chloride gradient, the radioactivity removed from O6-ethylguanine appeared in a low-density band. This material has been isolated and subjected to trypsin digestion and high-pressure liquid chromatography analysis; it was sensitive to trypsin and the digest contained new high-pressure liquid chromatography peaks characteristic of oligopeptides. Radioactive peaks from the trypsin digestion have been digested further to the amino acid level and have been shown to contain S-[3H]ethylcysteine. Thus, we conclude that the repair activity in rat liver chromatin removes the ethyl group from O6-ethylguanine and transfers it to a cysteine moiety contained in an acceptor protein.