Reaction of dithiothreitol and para-nitroacetophenone with different radical precursors of .OH radical-induced strand break formation of single-stranded DNA in anoxic aqueous solution.
The yields of single-strand breakage (ssb) in single-stranded calf thymus DNA (ssDNA) have been determined after 60Co gamma-irradiation of aqueous anoxic solutions in the presence of different concentrations of dithiothreitol (DTT), ascorbate or trans-4,5-dihydroxy-1,2-dithiane, using low-angle laser light scattering. The influence of DTT on the kinetics of ssb formation has been determined by conductivity measurements in pulse radiolysis. The results suggest that strand breakage in ssDNA proceeds via two modes of about equal contribution and with half-lives of about 7 ms and 0.8s, respectively. Both modes reflect reactions of at least two DNA radicals, which react with DTT by hydrogen-atom transfer reactions with similar rate constants of about 5-9 x 10(5) dm3 mol-1 s-1. These hydrogen-atom transfer reactions inhibit strand break formation. The slow mode is shown to represent the decay of base-radicals to generate sugar radicals. The involvement of the oxidizing .OH adduct radical of guanine in the formation of strand breaks can be ruled out and there is no evidence for a contribution from the anion or radical anion of DTT to the inhibition of strand breaks via electron transfer reactions to DNA radicals.
Max-Planck-Institut für Strahlenchemie, Mülheim a.d. Ruhr, Germany.
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Rao
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Bothe
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Schulte-Frohlinde
D000098:Acetophenones; D000818:Animals; D004249:DNA Damage; D004277:DNA, Single-Stranded / Q000528:radiation effects*; D004229:Dithiothreitol; D006878:Hydroxides; D017665:Hydroxyl Radical; D066298:In Vitro Techniques; D010100:Oxygen; D011838:Radiation-Sensitizing Agents; D012996:Solutions; D014867:Water
D000098:Acetophenones; D004277:DNA, Single-Stranded; D006878:Hydroxides; D011838:Radiation-Sensitizing Agents; D012996:Solutions; D014867:Water; C001972:4-nitroacetophenone; D017665:Hydroxyl Radical; D010100:Oxygen; D004229:Dithiothreitol