Monitoring the Structure-Dependent Reaction Pathways of Guanine Radical Cations in Triplex DNA: Deprotonation Versus Hydration.

Exposure of DNA to one-electron oxidants leads initially to the formation of guanine radical cations (G), which may degrade by deprotonation or hydration and ultimately cause strand breaks or 8-oxoG lesions. As the structure is dramatically changed by binding of the third strand in the major groove of the target duplex, it makes the triplex an interesting DNA structure to be examined and compared with the duplex on the G degradation pathways. Here, we report for the first time the time-resolved spectroscopy study on the G reaction dynamics in triplex DNA together with the Fourier transform infrared characterization of steady-state products, from which structural effects on the reactivity of G are unraveled. For an antiparallel triplex-containing GGC motif, G mainly suffers from fast deprotonation (9.8 ± 0.2) × 10 s, featuring release of both N-H and N-H of G in the third strand directly into bulk water. The much faster and distinct deprotonation behavior compared to the duplex should be related to long-resident water spines in the third strand. The G hydration product 8-oxoG is negligible for an antiparallel triplex; instead, the 5-HOO-(G-H) hydroperoxide formed after G deprotonation is identified by its vibrational marker band. In contrast, in a parallel triplex (CGC), the deprotonation of G occurs slowly (6.0 ± 0.3) × 10 s with the release of N-H, while G hydration becomes the major pathway with yields of 8-oxoG larger than in the duplex. The increased positive charge brought by the third strand makes the G radical in the parallel triplex sustain more cation character and prone for hydration. These results indicate that non-B DNA (triplex) plays an important role in DNA damage formation and provide mechanistic insights to rationalize why triplex structures might become hot spots for mutagenesis.