Death through respiratory failure of a fraction of ultraviolet-irradiated Escherichia coli b-r cells

Abstract

Escherichia coli B/r cells grown on a glycerol-containing medium and ultraviolet (UV)-irradiated to about 0.5% survival respire for about 1 hr and then cease for several hours. The cells that have completed repair and recovery processes begin to divide about 120 min after UV treatment, but this division is completely inhibited in liquid medium by caffeine, which delays repair of the irradiated deoxyribonucleic acid (DNA). When 5-fluorouracil (FU) is used to maintain respiration, the number of cells which form colonies when plated increases about 60-fold within 1 hr after irradiation. At least part of this increase does not involve repair while the cells are in the liquid medium because when caffeine is present there is still a 20-fold increase in colony formation. We conclude that many irradiated cells, although capable of carrying out complete and accurate repair of their DNA, die of respiratory failure; only when continuance of respiration is favored by FU treatment is their colony-forming potential realized. After an early increase, the number of cells able to form colonies in medium that contains FU remains constant while the completion of repair and recovery occurs. After these processes are completed, the number of cells able to form colonies increases slowly, except in the presence of caffeine, presumably because the late increase requires that repair steps take place while the cells are in liquid medium prior to cell division.