Analysis of the physiological control of replication of ColE1-type plasmids.
The physiology of ColE1-type plasmid replication in a growing host has been examined both theoretically, using computer simulation, and experimentally, by observing replication of the plasmid pBR322 after a nutritional shift-up from glycerol minimal medium (doubling time 71 min) to LB medium (doubling time 24 min). The theory was based on a negative control model and uses three rate equations: for the accumulation of cell mass, for the accumulation of the replication inhibitor, and for the rate of plasmid synthesis. The implications of the theory were explored by simulating the effects of changes in the expression of replication control genes. The nutritional shift-up experiment showed that plasmid replication was blocked immediately after the shift for about half a mass doubling time; after that time, replication rapidly increased until plasmid numbers per unit volume of culture parallelled the increase in culture mass. After the establishment of steady-state growth in the post-shift medium, the plasmid concentration (plasmids per cell mass) was reduced in comparison to pre-shift growth in the same proportion as the culture doubling time. The results showed that plasmid replication factors are under metabolic control and that the changes in the control of these factors compensate one another during steady-state growth, but not immediately after the medium shift.
D003086:Bacteriocin Plasmids; D003198:Computer Simulation; D004261:DNA Replication; D004269:DNA, Bacterial / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics*; D008957:Models, Genetic; D010957:Plasmids; D012313:RNA / Q000502:physiology; D012329:RNA, Bacterial / Q000502:physiology; D014158:Transcription, Genetic
D004269:DNA, Bacterial; C042027:RNA I; C034477:RNA primers; D012329:RNA, Bacterial; D012313:RNA