Formation of single- and double-strand breaks of pBR322 plasmid irradiated in the presence of scavengers

By the method of gel electrophoresis, radiation-induced DNA single- and double-strand breaks (SSB, DSB) were studied with a model system of pBR322 solution in vitro in the presence of ^·OH radical scavengers, mannitol and TE (10^–2 mol dm^–3 Tris and 10^–3 mol dm^–3 ethylene diamine tetra-acetic acid). Experiments showed that SSB resulted from one-hit events of radiation energy deposition and DSB resulted from both one-hit and two-hit energy deposition events and so were distinguished into two classes of αDSB and βDSB. Moreover, α/β, where α is the number of DSB per unit dose induced in one irradiation event and β the number of DSB per unit squared dose induced by the combination of two independent SSB, was related to the scavenging capacity, σ, and for σ>10⁸ s^–1,αDSB predominate over DSB. On the other hand, if σ<2×10⁸ s^–1, the measured G(αDSB) decreased in parallel with G(SSB), i.e., G(αDSB)/G(SSB) was a constant. When σ>2×10⁸ s^–1, G(αDSB) decreased slightly so that the ratio of αDSB to SSB evidently increased. Therefore, αDSB could be induced by the radical transfer mechanism for σ<2×10⁸ s^–1 and contrarily produced by the local multiply damaged sites (LMDS) mechanism for σ larger than this value. In addition, the distance for two independent complementary SSB forming a DSB was deduced, but no apparent variation of it was found in the wide σ range from ∼10⁵ to ∼10⁹ s^–1, which shows that the DNA steric structure was not influenced by mannitol. Received: 28 September 1998 / Accepted in revised form: 20 March 1999