Etheno (epsilon) and related DNA adducts are formed from the reaction of certain bifunctional electrophiles with DNA. Our interest has been focused on oxiranes substituted with leaving groups, e.g. 2-chlorooxirane, the epoxide derived from the carcinogen vinyl chloride. The chemical mechanisms of the formation of the major etheno products derived from adenine, cytosine and guanine have been elucidated by nuclear magnetic resonance analysis and 13C-labelled precursors. The amounts of all major etheno adducts have been quantified in DNA treated with 2-chlorooxirane by coupled high-performance liquid chromatography of nucleoside and base products. 1,N2-epsilon-Gua, its formally hydrated but stable hemiaminal HO-ethanoGua (5,6,7,9-tetrahydro-7-hydroxy-9-oxoimidazo[1,2-a]purine) and 1,N2-ethanoGua have all been inserted at a single site in oligonucleotides. All three of these bases block polymerases, cause misincorporations and produce some mutations in bacteria. The patterns of blockage and substitution vary among polymerases. In nucleotide excision repair-deficient Escherichia coli, 1,N2-epsilon-Gua yielded a calculated 16% mutation frequency (base-pair substitutions) when the results were corrected for strand usage. 1,N2-epsilon-Gua was also examined in Chinese hamster ovary cells with a stable integration system; the mutants are more complex than observed in bacteria and include rearrangements, deletions and base-pair substitutions other than at the adduct site.