Evaluation of lesion clustering in irradiated plasmid DNA

2005, Vol. 81, No. 1 , Pages 41-54 (doi:10.1080/09553000400017895)
1Dept. of Particle Physics, Weizmann Institute of Science, Rehovot, 76100, Israel
2Dept. of Biological Chemistry, Weizmann Institute of Science, Rehovot, 76100, Israel
3Dept. of Radiation Medicine, Loma Linda University Medical Center, Loma Linda, CA 92354, USA
4Dept. of Radiology, University of California at San Diego, La Jolla, CA 92093, USA
5Physikalisch Technische Bundesanstalt, D-38116 Braunschweig, Germany
Address for correspondence: C. Leloup. Dept. of Particle Physics, Weizmann Institute of Science, Rehovot, 76100, Israel, Currently at the center for Radiological Research, Columbia University, 630W 168th street VC 11-219, NY 10034, USA



Purpose: To measure the yield of DNA strand breaks and clustered lesions in plasmid DNA irradiated with protons, helium nuclei, and γ-rays.

Materials and methods: Plasmid DNA was irradiated with 1.03, 19.3 and 249 MeV protons (linear energy transfer = 25.5, 2.7, and 0.39 keV μm – 1 respectively), 26 MeV helium nuclei (25.5 keV μm) and γ-rays (137Cs or 60Co) in phosphate buffer containing 2 mM or 200 mM glycerol. Single-and double-strand breaks (SSB and DSB) were measured by gel electrophoresis, and clustered lesions containing base lesions were quantified by converting them into irreparable DSB in transformed bacteria.

Results: For protons, SSB yield decreased with increasing LET (linear energy transfer). The yield of DSB and all clustered lesions seemed to reach a minimum around 3 keV μm – 1. There was a higher yield of SSB, DSB and total clustered lesions for protons compared to helium nuclei at 25.5 keV μm – 1. A difference in the yields between 137Cs and 60Co γ-rays was also observed, especially for SSB.

Conclusion: In this work we have demonstrated the complex LET dependence of clustered-lesion yields, governed by interplay of the radical recombination and change in track structure. As expected, there was also a significant difference in clustered lesion yields between various radiation fields, having the same or similar LET values, but differing in nanometric track structure.