Topics in Free Radical-mediated DNA Damage: Purines and Damage Amplification—superoxic Reactions—bleomycin, the Incomplete Radiomimetic

Abstract

Only a small percentage of the DNA damage set by ionizing radiation in the living cell manifests itself as lethal. It is now increasingly accepted that clustered lesions may constitute the kind of damage that the repair enzymes cannot adequately deal with. The question is raised as to whether damage amplification reactions (radical transfer reactions) may contribute to these clustered lesions, and examples of such damage amplification reactions are given. In one example a purine is involved. With 2′-deoxy adenosine and 2′-deoxy guanosine it is shown that these purine nucleosides undergo unexpected radical reactions. Evidence for the radical transfer from the purine to the sugar moiety is provided by the formation of the 5′-aldehydes. These products have been assayed with 2-thiobarbituric acid (TBA), a reagent commonly applied to the detection of malonaldehyde. TBA-reactive material has also been assayed in γ-irradiated and bleomycin-treated DNA. In γ-irradiated DNA, about one-third of this is free malonaldehyde, while the major part of the TBA-reactive material remains bound to the DNA. In contrast, bleomycin-treated DNA yields practically no free malonaldehyde, and the major TBA-reactive products are identified as the thymine and adenine base propenals. There is also further unidentified low-molecular-weight, TBA-reactive material. It is concluded that the drug must be involved not only in the first step (H-abstraction from the sugar moiety), but also in the subsequent free-radical reactions on the way to the base propenals.