DNA Damage Produced by Exposure of Supercoiled Plasmid DNA to High- and Low-LET Ionizing Radiation: Effects of Hydroxyl Radical Quenchers

Abstract

Production of DNA damage by exposure to ionizing radiation was measured in two in vitro systems. A supercoiled plasmid of 7·3 kbp was isolated and exposed in an aqueous environment to ⁶⁰Co γ rays and JANUS 0·85 MeV fission-spectrum neutrons. Dose responses for the production of single-strand breaks (ssbs) and double-strand breaks (dsbs) were computed from the conversion of the supercoil to its relaxed and linear forms. The relative effectiveness (neutrons:γ-rays) for destruction of genetic transforming activity of M13 viral DNA was 0·23, close to that for ssb production, in contrast with the situation for biological effects such as lethality, mutagenesis and cellular transformation measured in mammalian cells, where RBEs are > 1. The role of hydroxyl (OH) radicals in DNA damage induction by neutrons was investigated by exposure of plasmid in the presence of known quenchers of this species. Of four quenchers tested, all were able to reduce the yields of both ssbs and dsbs. These findings are consistent with a model for ssb and dsb induction by high linear energy transfer radiation that involves OH radical mediation.