Mutagenic specificity of a derivative of 3-nitrobenzanthrone in the supF shuttle vector plasmids

3-Nitrobenzanthrone (NBA) is a powerful bacterial mutagen and a suspected human carcinogen present in diesel exhaust and airborne particulates (Enya, T., et al. (1997) Environ. Sci. Technol. 31, 2772-2776]. In the accompanying paper [Enya, T,, ct al. (1998) Chem. Res. Toxcol. 11, 1460-1467], N-acetoxy-N-acetyl-3-aminobenzanthrone (N-Aco-N-Ac-ABA) was synthesized to yield the DNA adducts of NEA. In this work, to investigate the mutagenic specificity of NBA in human cells, we analyzed mutations induced by N-Aco-N-Ac-ABA using the supF shuttle vector plasmids. Base sequence analysis of 110 and 100 plasmids with mutations in the supF gene propagated in normal cells [WI38-VA13] and nucleotide excision repair deficient cells [XP2OS(SV)], respectively; revealed that the majority of the mutations were base substitutions (85 and 90%) and the rest were deletions and insertions (10 and 15%) in both cell lines. About half of the mutant. plasmids had a single base substitution. Of the base substitutions, the most frequent mutation was G.C to T.A transversion (41 and 51%), Followed by G.C to A.T transitions (18 and 24%) in either cell. The mutations were distributed not randomly but located at several hot spots, and almost all (nine of ten) hot spots were at the sites of G.C base pairs. The polymerase stop assay in the supF gene revealed that N-Aco-N-Ac-ABA preferentially bound to guanine residues, and mutation sites were generally consistent with the sites where the guanine adducts were formed.