Purpose: It was previously shown that gamma-irradiation of aqueous solution
s of plasmid DNA in the presence of millimolar concentrations of thiocyanat
e ions leads to the formation in very high yields of sites recognized by th
e base excision repair endonuclease formamido-pyrimidine-DNA N-glycosylase
(FPG). The authors wished to characterize the mechanism responsible for the
production of these FPG-sensitive sites.
Materials and methods: An aqueous solution of plasmid DNA containing thiocy
anate ions was irradiated with Cs-137 gamma-rays After irradiation, aliquot
s were treated with FPG. Break yields were determined using neutral agarose
gel electrophoresis.
Results: The yield of FPG-sensitive sites decreased with decreasing enzyme
activity, increasing thiocyanate concentration, increasing dose-rate, incre
asing ionic strength; increasing nitrite or iodide concentration, and decre
asing oxygen concentration.
Conclusion: The observations suggest that the monomeric thiocyanate radical
SCN. is an intermediate in the reaction, and that the yields of FPG-sensit
ive sites are determined by competition between the disproportionation of t
he dimeric radical anion (SCN)(2)(.-) and the fate of a one-electron oxidiz
ed guanine species in DNA. The latter can react with oxygen to produce an F
PG-sensitive site or can be reduced without producing an FPG-sensitive site
. The results help to clarify the mechanisms responsible for DNA damage by
the direct effect of ionizing radiation.