Loss of photoreversibility for UV mutation in E. coli using 405 nm or near-UV challenge

E. coli mutagenized with germicidal ultraviolet light (UV) were incubated to allow for development of mutation-fixation processes. Fixation was estimated from the effects on mutation frequency of photoreactivation challenge during the first 60 min post-UV. Two different light sources were used for photoreactivation, one providing effective light primarily at 405 nm and another providing a broad range of near-UV around 365 nm. Kinetics for the loss of photoreversibility (LOP) were determined. The times for completion of LOP in wild-type cells indicated one fixation process for back mutation and another for de novo or converted suppressor mutation regardless of the light source. Using 405-nm light for photoreactivation, the LOP kinetics for back mutation and de novo suppressor mutation in uvrA cells were similar. Hence, classical observations were confirmed here. Immediately post-UV all mutation frequencies were more sensitive to near-UV than 405-nm light. Experiments with rel cells supported the idea that growth delay and inhibition of induced lexA-coordinated responses may be responsible for this early, pronounced sensitivity to photoreactivation by near UV. For back mutation and de novo suppressor mutation, the sensitivity to 405-nm light was initially small and actually increased for 10-15 min. Possibly genome conformation changes are induced by UV and this affects the efficiency of photoenzymatic monomerization of 405-nm light during the first 10-15 min after irradiation.