Heavy Ion-Induced DNA Double-Strand Breaks in Yeast

Abstract Kiefer, J., Egenolf, R. and Ikpeme, S. Heavy Ion-Induced DNA Double-Strand Breaks in Yeast. Radiat. Res. 157, 141–148 (2002). Induction of DSBs in the diploid yeast, Saccharomyces cerevisiae, was measured by pulsed-field gel electrophoresis (PFGE) after the cells had been exposed on membrane filters to a variety of energetic heavy ions with values of linear energy transfer (LET) ranging from about 2 to 11,500 keV/μm, 241Am α particles, and 80 keV X rays. After irradiation, the cells were lysed, and the chromosomes were separated by PFGE. The gels were stained with ethidium bromide, placed on a UV transilluminator, and analyzed using a computer-coupled camera. The fluorescence intensities of the larger bands were found to decrease exponentially with dose or particle fluence. The slope of this line corresponds to the cross section for at least one double-strand break (DSB), but closely spaced multiple breaks cannot be discriminated. Based on the known size of the native DNA molecules, breakage cross sections per base pair were calculated. They increased with LET until they reached a transient plateau value of about 6 × 10–7 μm2 at about 300–2000 keV/μm; they then rose for the higher LETs, probably reflecting the influence of δ electrons. The relative biological effectiveness for DNA breakage displays a maximum of about 2.5 around 100–200 keV/μm and falls below unity for LET values above 103 keV/μm. For these yeast cells, comparison of the derived breakage cross sections with the corresponding cross section for inactivation derived from the terminal slope of the survival curves shows a strong linear relationship between these cross sections, extending over several orders of magnitude.