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Temperature-Sensitive Recovery of a Mutant of Escherichia coli K-12 Irradiated with Ultraviolet Light

Biochemistry Laboratory, Department of Chemistry, National Institute of Radiological Sciences, Anagawa Chiba-shi, Japan

ABSTRACT

URT-43 is a mutant of Escherichia coli K-12 which gives a much larger number of survivors when ultraviolet (UV)-irradiated bacteria are incubated on agar medium at 30 C than when they are incubated on the medium at 41 C, although in both cases the number of survivors is fewer than that given by its wild-type ancestor. The UV sensitivity of this mutant was found to be markedly influenced by the presence of a high concentration of NaCl or sucrose in the plating medium. Thus, when irradiated bacteria were plated on agar medium containing 2% NaCl or 0.5 M sucrose at 30 C, they exhibited a resistance similar to that of their wild-type ancestor. At 30 C, there was also an extensive recovery in liquid M9 medium supplemented with all of the nutrients required for growth and NaCl or sucrose. At 41 C, however, the recovery was greatly inhibited. Direct chemical analysis of thymine dimers has revealed that no significant amount of the dimer was released from deoxyribonucleic acid during the period of extensive recovery. It was concluded, therefore, that the temperature-sensitive recovery of URT-43 does not accompany excision of the bulk of pyrimidine dimers. To learn the gene function involved in the recovery, double mutants carrying an additional mutation either in a uvr or a rec gene have been investigated for their UV sensitivities and recovery in liquid medium. It was found that recA^– and recB^– derivatives retain the ability of undergoing an efficient recovery at a low temperature, whereas uvrB^– and uvrC^– derivatives have completely lost this ability. For these reasons, it was concluded that the mechanism responsible for the recovery of URT-43 involves the function controlled by the uvr genes. The results of photoreactivation suggested that most of the entities dealt with during recovery were pyrimidine dimers.