SOLUTION STRUCTURE OF THE AMINOFLUORENE [AF]-INTERCALATED CONFORMER OF THE SYN-[AF]-C-8-DG ADDUCT OPPOSITE DC IN A DNA DUPLEX

We report below on a conformational equilibrium between AF-intercalate d and AF-external states in slow exchange for the [AF]dG lesion positi oned opposite dC in the d(C-[AF]G-C) d(G-C-G) sequence context, The sl ow exchange between states is attributed to interconversion between sy n glycosidic torsion angle in the AF-intercalated and anti torsion ang le in AF-external conformers of the [AF]dG opposite dC containing dupl ex. The present paper describes an NMR-molecular mechanics study that defines the solution structure of the AF-intercalated conformer for th e case of [AF]dG adduct positioned opposite dC in the d(C-[AF]G-C).d(G -C-G) sequence context. The structure is of the base displacement-inte rcalation type where the aminofluorene ring is intercalated into the h elix between intact Watson-Crick dG.dC base pairs, which results in a displacement of the modified guanine ring into the major groove where it stacks with the major groove edge of its 5'-flanking cytosine in th e adduct duplex. The conformational equilibrium between AF-intercalate d conformer (similar to 70%) with a syn alignment and AF-external conf ormer (similar to 30%) with an anti alignment for the [AF]dG adduct po sitioned opposite dC in the d(C-[AF]G-C).d(G-C-G) sequence context can be contrasted with our earlier demonstration that the population is 1 00% for the AP-intercalated conformer with a syn alignment at the N-(d eoxyguanosin-8-yl)-2-aminopyrene ([AP]dG) adduct site positioned oppos ite dC in the same sequence context [Mao, B,, Vyas, R. R., Hingerty, B , E,, Broyde, S,, Basu, A, K., and Patel, D, J. (1996) Biochemistry, 3 5, 12659-12670]. This shift in population may reflect the much larger size of the pyrenyl ring of the [AP]dG adduct compared to the fluoreny l ring of the [AF]dG adduct which in turn might provide for a greater overlap of the aromatic amine with the flanking base pairs in the inte rcalated conformer of the former adduct in DNA.