Tuning the replication fork progression by the initiation frequency

Abstract

The thermo-resistant period of the thermo-sensitive ribonucleotide reductase RNR101 encoded by the nrdA101 allele in Escherichia coli is prolonged for 50 min at 42°C, enabling an increase in DNA content of about 45%. Assuming that fork progression in the nrdA101 mutant is impaired, the question whether reduced number of ongoing replication rounds altered the thermo-resistant period in this strain was investigated. Decreases in the oriC/terC ratio and in the number of oriC per cell at 30°C were found in the presence of oriC228, oriC229 and oriC239 alleles in strain nrdA101. Correlated with this effect, increased thermo-resistance period of the RNR101 was allowed, and the detrimental effects on cell division, chromosome segregation and cell viability observed in the nrdA101 mutant at 42°C were suppressed. These results indicate that conditions leading to chromosome initiation deficiency at 30°C enhance the replication fork progression in the nrdA101 mutant at 42°C. We propose that coordination between initiation frequency and replication fork progression could be significant for most of the replication systems with important consequences in their cell cycle regulation.