The major, N-2-dG adduct of (+)-anti-B[a]PDE induces G -> A mutations in a5 '-A(G)under-bar-A-3 ' sequence context

Previously, in a random mutagenesis study, the (+)-anti diol epoxide of ben zo[a]pyrene [(+)-anti-B[a]PDE] was shown to induce a complex mutational spe ctrum in the supF gene of an Escherichia coli plasmid, which included inser tions, deletions and base substitution mutations, notably a significant fra ction of GC-->TA, GC-->AT and GC-->CG mutations. At some sites, a single ty pe of mutation dominated and to understand individual mutagenic pathways th ese sites were chosen for study by site-specific means to determine whether the major adduct, [+ta]-B[a]P-N-2-dG, was responsible. [+ta]-B[a]P-N2-dG w as shown to induce similar to 95% G-->T mutations in a 5'-T (G) under bar C -3' sequence context and similar to 80% G-->A mutations in a 5'-C (G) under bar T-3' sequence context. (+)-anti-B[a]PDE induced principally GC-->CG mu tations in the G133 sequence context (5'-A (G) under bar A-3') in studies u sing both SOS-uninduced or SOS-induced E.coli. Herein, [+ta]-B[a]P-N2-dG is shown to induce principally G-->A mutations (>90%) either without or with SOS induction in a closely related 5'-A (G) under bar A-3' sequence context (identical over 7 bp), This is the first time that there has been a discre pancy between the mutagenic specificity of(+)-anti-B[a]PDE versus [+ta]-B[a ]P-N2-dG. Eight explanations for this discordance are considered. Four are ruled out; e.g. the second most prevalent adduct [ + ca]-B[a]P-N2-dG also i nduces a preponderance of G-->A mutations (>90%), so it also is not respons ible for (+)-anti-B[a]PDE-induced G133-->C mutations. The four explanations not ruled out are discussed and include that another minor adduct might be responsible and that the 5'-A (G) under bar A-3' sequence context differed slightly in the studies with [+ta]-B[a]P-N2-dG versus(+)-anti-B[a]PDE, In spite of the discordance, [Sta]-B[a]P-N-2-dG induces G-->A mutations in the context studied herein and this result has proven useful in generating a h ypothesis for what conformations of [+ta]-B[a]P-N-2-dG are responsible for G-->T versus G-->A mutations.