Comparison of Sensitivity of Rad Mutants of Diploid Yeast to Heat and Gamma Radiation: Cellular Target for Heat Inactivation

Summary

Wild type and radiation-sensitive mutants rad 53, 54 and 55 of the diploid yeast Saccharomyces cerevisiae, in stationary and log phase were exposed to gamma radiation and hyperthermia (51°C) in order to compare their sensitivity to these agents. The wild type diploid strain exposed to gamma rays showed a sigmoidal survival curve both in stationary and log phase cultures. Log phase cells were significantly more resistant than stationary phase cells. When compared to wild type, the gamma radiation response of the mutants indicated that the mutations in these RAD loci render the cells sensitive in stationary phase and very sensitive in log phase. The response of mutants to hyperthermia was similar to that of wild type cells in both the phases. The log phase cells of both wild type and mutants were more sensitive to heat than stationary phase cells. These results suggest that the RAD loci are not involved in the repair of hyperthermic damage. Since it is known that the products of the RAD genes are involved in the repair of DNA damage, the wild type response of these rad mutants to hyperthermia indicates that the DNA may not be the principal target for hyperthermic killing. Furthermore, the enhanced thermal sensitivity of log phase cells, containing higher amounts of active enzymes and sensitive membrane, strongly suggests that proteins and/or membranes could be the primary targets for thermal inactivation.