DNA strand break yields after post-high LET irradiation incubation with endonuclease-III and evidence for hydroxyl radical clustering

Abstract

Purpose : To determine the increase in single- (SSB) and double-strand break (DSB) yields after post-high LET irradiation incubation of plasmid DNA with the endonuclease-III (endo-III) of Escherichia coli. Materials and methods : Plasmid DNA in aerobic aqueous solution was irradiated with one of five radiation types: 137 Cs γ-rays (LET ~0.3keV μ m -1) , 244 Cm α -particles (140-190 keV μ m -1) , 4 He ions (97 keV μ m -1) , 56 Fe ions (143keV μ m -1) or 197 Au ions (1440keV μ m -1) . The irradiated samples were then incubated with endo-III. SSB and DSB yields were quantified by agarose gel electrophoresis. Results : Endo-III incubation produced an increase in the SSB and DSB yields. The increases were in general lower after the high LET irradiation than after γ-irradiation. This may reflect inhibition of the activity of endo-III by the nearby DNA damage expected from high LET radiation. It can be shown that even if the activity of endo remains unchanged, significantly lower increases in SSB and DSB yields would still be expected. Conclusion : The results provide evidence for clustered DNA damage after high LET irradiation.