Purpose: It has been previously argued that ii-irradiation of plasmid DNA i
n the presence of thiocyanate ions produces products recognized by the E. c
oli base excision-repair endonuclease formamidopyrimidine-DNA N-glycosylase
(FPG), and there that derive from an intermediate guanyl radical species.
The wish was to characterize the reactivity of this intermediate with reduc
ing agents.
Materials and methods: Aqueous solutions of plasmid DNA containing either b
romide or thiocyanate (10(-3) to 10(-1) mol dm(-3)) and also one of six oth
er additives (azide, ferrocyanide, iodide, nitrile, promethazine, tryptopha
n, 10(-7) to 10(-3) mol dm(-3)) were subjected to Cs-137 gamma -irradiation
(662 keV). After irradiation, the plasmid was incubated with FPG. Strand b
reak yields before and after incubation were determined by agarose gel elec
trophoresis under neutral conditions.
Results: The very high yields of FPG-sensitive sites in the presence of SCN
- or Br- decreased significantly with increasing concentrations of all of t
he sis additives, with promethazine and tryptophan being the most efficient
additives, and azide and iodide the least.
Conclusions: From the results it is possible to estimate values of the rate
constants for the reduction of the DNA guanyl radical (5 x 10(5), 2 x 10(5
), 10(7) and 10(7) dm(3) mol(-1) s(-1) for ferrocyanide, nitrite, promethaz
ine and tryptophan respectively).