Mechanism of oxidative DNA damage induction in a strict anaerobe, Prevotella melaninogenica

We investigated the mechanism of the oxidative DNA damage induction by exposure to O(2) in Prevotella melaninogenica, a strict anaerobe. Flow cytometry with hydroethidine and dichlorofluorescein diacetate showed that O(2) exposure generated O(2)*-) and H(2)O(2). Results of electron spin resonance with alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone and ethanol showed that O(2) exposure also induced *OH radical generation in P. melaninogenica loaded with FeCl(2) but not in samples without FeCl(2) loading. In P. melaninogenica, O(2) exposure increased 8-hydroxydeoxyguanosine (8OHdG), typical of oxidative DNA damage. Catalase inhibited the increase, but the *OH radical scavengers did not. Phenanthroline, a membrane-permeable Fe and Cu chelator, increased the 8OHdG induction. In FeCl(2)-loaded samples, induction of 8OHdG decreased. Addition of H(2)O(2) markedly increased 8OHdG levels. These results indicate that in P. melaninogenica, exposure to O(2) generated and accumulated O(2)* and H(2)O(2), and that a crypto-OH radical generated through H(2)O(2) was the active species in the 8OHdG induction.