No abstract

Induction of DNA double-strand breaks in diploid wild-type yeast cells, and inactivation of diploid mutant cells (rad54-3) unable to repair DNA double-strand breaks, were studied with aluminium K (1.5 keV) and carbon K (0.278 keV) characteristic X-rays. The induction of DNA double-strand breaks was found to increase linearly with absorbed dose for both characteristic X-rays. Carbon K X-rays were more effective than aluminium K X-rays. Relative to 60Co gamma-rays the r.b.e.-values for the induction of DNA double-strand breaks were found to be 3.8 and 2.2 for carbon K and aluminium K X-rays respectively. The survival curves of the rad54-3 mutant cells were exponential for both ultrasoft X-rays. For inactivation of rad54-3 mutant cells, the r.b.e.-values relative to 60Co gamma-rays were 2.6 and 2.4 for carbon K and aluminium K X-rays, respectively. The DNA double-strand break data obtained with aluminium K and carbon K X-rays are in agreement with the data obtained for gene mutation, chromosome aberrations and inactivation of mammalian cells, suggesting that DNA double-strand breaks are the possible molecular lesions leading to these effects.

DNA double-strand breaks (DSBs) are 2.9 times more frequently induced in yeast cells exposed to sparsely ionizing 30-MeV electrons under oxic compared to anoxic conditions. The rejoining of DSBs induced under anoxic conditions was investigated under conditions allowing repair of potentially lethal damage and compared to the rejoining of DSBs induced in oxic cells. In contrast to the biphasic rejoining kinetics of DSBs induced in oxic cells, the rejoining kinetics of DSBs induced in anoxic cells is complicated by the formation of secondary DSBs. These arise during postirradiation incubation of cells, presumably as a consequence of repair processes acting on radiation-induced lesions other than DSBs. These secondary DSBs may at least partially explain the finding that a greater fraction of unrejoinable DSBs is present in cells irradiated under anoxic compared to oxic conditions. As a consequence, the oxygen enhancement ratio of the yield of the remaining DSBs is decreasing in the course of DSB rejoining.

Effects of 2-deoxy-D-glucose (2-DG) on radiation-induced DNA double-strand breaks (dsb) have been studied under non-growth conditions in a respiratory-deficient strain of the yeast Saccharomyces cerevisiae. Velocity sedimentation in neutral sucrose gradients was used to measure DNA dsb. Addition of 2-DG to the liquid-holding medium (67 mM phosphate buffer, pH 5, 30 degrees C) at an equimolar concentration with glucose (50 mM) reduced the rate and extent of dsb rejoining. The inhibition of rejoining mediated by 2-DG is reversible for the majority--but not all--of the radiation-induced dsb.

DNA double-strand break (dsb) induction in diploid yeast was measured by neutral sucrose sedimentation after exposure to very heavy ions with values of linear energy transfer (LET) ranging from about 300 to 11500 ke V/microns. Linear fluence dependencies were found in all cases from which dsb production cross-sections (sigma dsb) could be calculated. Corresponding cross-sections for cell killing (sigma i) were derived from final slopes of survival curves measured in parallel and for the same fluence range. A close correlation was found between sigma i and sigma dsb. It is calculated that over the entire LET range, including 30 MeV electron irradiation, about 22 dsb are induced per lethal event when high exposures are considered.