Characterization of a novel reaction by human O6-alkylguanine-DNA alkyltransferase with 1,3-bis(2-chloroethyl)-1-nitrosourea-treated DNA

Abstract

The reaction of partially purified human O6-alkylguanine-DNA alkyltransferase with 1,3-bis(2-chloroethyl)-1-nitrosourea-treated DNA produces a covalent DNA-protein complex. Evidence that the complex-forming and the alkyltransferase activities are one and the same includes their copurification, identical heat inactivation kinetics, and similar responses to inhibitors. This novel reaction thus provides a sensitive alternative assay for detecting the alkyltransferase. It also provides support for a model in which N1,O6-ethanoguanine is an intermediate in formation of chloroethylnitrosourea-induced DNA interstrand cross-links.