Abstract

Bacterial chromosomes harbour a unique origin of bidirectional replication, oriC. They are almost always circular, with replication terminating in a region diametrically opposite to oriC, the terminus. The oriC-terminus organisation is reflected by the orientation of the genes and by the disposition of DNA-binding protein motifs implicated in the coordination of chromosome replication and segregation with cell division. Correspondingly, the E. coli and B. subtilis model bacteria possess a replication fork trap system, Tus/ter and RTP/ter, respectively, which enforces replication termination in the terminus region. Here, we show that tus and rtp are restricted to four clades of bacteria, suggesting that tus was recently domesticated from a plasmid gene. We further demonstrate that there is no replication fork system in Vibrio cholerae, a bacterium closely related to E. coli. Marker frequency analysis showed that replication forks originating from ectopic origins were not blocked in the terminus region of either of the two V. cholerae chromosomes, but progressed normally until they encountered an opposite fork. As expected, termination synchrony of the two chromosomes is disrupted by these ectopic origins. Finally, we show that premature completion of the primary chromosome replication did not modify the choreography of segregation of its terminus region.

Highlights

  • Genome architecture is extremely conserved in bacteria

  • The replication fork trap (RFT) effect on replication fork progression was monitored in E. coli using Marker Frequency Analysis (MFA) in strains harbouring an additional ectopic origin: in presence of an RFT, the convergence between the two forks continued to be in the Ter region, while in its absence, the convergence point was displaced to the midpoint between the two origins[36,49]

  • Our results unambiguously demonstrate that no replication fork trap (RFT) exists in V. cholerae

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Summary

OPEN Replication termination without a replication fork trap

Elisa Galli[1], Jean-Luc Ferat[1,2], Jean-Michel Desfontaines[1], Marie-Eve Val 3,4, Ole Skovgaard 5, François-Xavier Barre1 & Christophe Possoz[1]. S95, the length of the region centred on the fcp within which 95% of forks converged, increased from 4 kbp for the fcp[1] of EPV50 to 128 kbp for the fcpL of EGV140, 628 kpb for the fcpR of EGV140, 396 kbp for the fcpL of EGV111 and 176 kbp for the fcpR of EGV111 (Fig. 2 and Table S4) Together, these results suggest that replication is slightly perturbed when ectopic origins were added. Both fcp were located close to their corresponding midpoint, at less than 1.5% of each replicated DNA segment (Fig. 3B,C, Table S5) These results show that replication forks may progress beyond dif[2] over ~67% of the left replichore of chr[2] and over the entire right replichore of chr[2].

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