DNA damage in deoxynucleosides and oligonucleotides treated with peroxynitrite

Peroxynitrite (ONOO-) is a powerful oxidizing agent that forms in a reactio n of nitric oxide (NO.) and superoxide (O-2(-.)) We have investigated ONOO- -induced DNA damage using deoxynucleosides and oligonucleotides as model su bstrates, with particular attention paid to the oxidation of 8-oxodG by ONO O-. With regard to deoxynucleosides, ONOO- was found to have significant re activity only with dG; dA, dC, and dT showed minimal reactivity. However, t wo of the major products of ONOO--induced oxidation of dG (8-oxodG and 8-ni troG) were both found to be significantly more reactive with ONOO- than wit h dG. In the context of an oligonucleotide, we observed a concentration-dep endent oxidation of 8-oxodG to at least two types of products, one appearin g at ONOO- concentrations of less than or equal to 100 mu M and the other a t concentrations of greater than or equal to 500 mu M. We also examined the susceptibility of these oxidation products to repair by FaPy glycosylase, endonuclease III, uracil glycosylase, and MutY. FaPy glycosylase, which rec ognizes 8-oxoG as its primary substrate, was the only enzyme that exhibited an efficient reaction with 8-oxodG oxidation products at low ONOO- concent rations (less than or equal to 100 mu M); the product(s) formed at ONOO- co ncentrations of greater than or equal to 500 mu M either was not recognized or was poorly repaired by the enzymes. While processing of the lesions was inefficient with endonuclease III and not apparent with uracil glycosylase , the excision of A opposite an 8-oxoG lesion by the enzyme MutY was not af fected by the reaction of 8-oxoG; with ONOO-. In addition to demonstrating the complexity of ONOO- DNA damage chemistry, these results suggest that 8- oxodG may be a primary target of ONOO- in DNA.