Evidence that ultraviolet light-induced DNA replication death of recA bacteria is prevented by protein synthesis in repair-proficient bacteria.
The ultraviolet light (UV) survival curve of Escherichia coli WP10 recA trp is almost biphasic, with a greatly reduced shoulder but demonstrating a transition to a decreased slope with increasing fluences, indicating the presence in the culture of a low frequency of resistant cells. Treatment of the culture with chloramphenicol before UV exposure brought almost all of the cells to a high degree of UV resistance, by bringing them to the end of their DNA replication cycle. The survival curves of the repair-proficient E. coli WP2 trp showed a similar pattern with chloramphenicol treatment or tryptophan starvation before UV exposure, but only if protein synthesis were blocked by chloramphenicol for 60 min after UV exposure. The results suggest that when recA/lexA-regulon induction is prevented, either by the recA mutation or by inhibition of protein synthesis after UV exposure, death occurs unless the cells are in the resistant state characteristic of bacteria at the end of their DNA replication cycle. With repair-proficient bacteria treated before UV exposure with chloramphenicol, when protein synthesis is not blocked after UV exposure, a marked expansion of the shoulder occurs because of the function of another resistance-conferring mechanism. This mechanism also depends on the recA+ gene since expansion of the shoulder does not occur in recA bacteria when protein synthesis is inhibited before UV exposure.
Doudney
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Wadsworth Center for Laboratories and Research, New York State Department Health, Albany, NY 12201.
Rinaldi
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D002470:Cell Survival; D002701:Chloramphenicol / Q000494:pharmacology*; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics* / Q000528:radiation effects; D009154:Mutation; D011938:Rec A Recombinases / Q000235:genetics*; D014466:Ultraviolet Rays
D002701:Chloramphenicol; D011938:Rec A Recombinases