Spectroscopic and theoretical insights into sequence effects of aminofluorene-induced conformational heterogeneity and nucleotide excision repair

Časopis: BIOCHEMISTRY 46, 11263-11278
Autoři: Meneni, SR., Shell, SM., Gao, L., Jurecka, P., Lee, W., Sponer, J., Zou, Y., Chiarelli, MP., Cho, BP.
Rok: 2007

Abstrakt

A systematic spectroscopic and computational study was conducted in order to probe the influence of base sequences on stacked (S) versus B-type (13) conformational heterogeneity induced by the major dG adduct derived from the model carcinogen 7-fluoro-2-antinofluorene (FAF). We prepared and characterized eight 12-mer DNA duplexes (-AG*N-series, d[CTTCTAG*NCCTC]; -CG*N-series, d[CTTCTCG*NCCTC]), in which the central guanines (G*) were site-specifically modified with FAF with varying flanking bases (N = G, A, C, T). S/B heterogeneity was examined by CD, UV, and dynamic F-19 NMR spectroscopy. All the modified duplexes studied followed a typical dynamic exchange between the S and B conformers in a sequence dependent manner. Specifically, purine bases at the 3'-flanking site promoted the S conformation (G >A >C >T). Simulation analysis showed that the S/B energy barriers were in the 14-16 kcal/mol range. The correlation time's (iota = l/k) were found to be in the millisecond range at 20 degrees C. The van der Waals energy force field calculations indicated the importance of the stacking interaction between the carcinogen and neighboring base pairs. Quantum mechanics calculations showed the existence of correlations between the total interaction energies (including electrostatic and solvation effects) and the S/B population ratios. The S/B equilibrium seems to modulate the efficiency of Escherichia coli UvrABC-based nucleotide excision repair in a conformation-specific manner: i.e., greater repair susceptibility for the S over B conformation and for the -AG*N- over the -CG*N- series. The results indicate a novel structure-function relationship, which provides insights into how bulky DNA adducts are accommodated by UvrABC proteins.