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Chromosomal inheritance in bacteria usually entails bidirectional replication of a single chromosome from a single origin into two copies and subsequent partitioning of one copy each into daughter cells upon cell division. However, the human pathogen  Vibrio cholerae  and other  Vibrionaceae  harbor two chromosomes, a large Chr1 and a small Chr2. Chr1 and Chr2 have different origins, an  oriC- type origin and a P1 plasmid-type origin, respectively, driving the replication of respective chromosomes. Recently, we described naturally occurring exceptions to the two-chromosome rule of  Vibrionaceae : i.e., Chr1 and Chr2 fused single chromosome  V. cholerae  strains, NSCV1 and NSCV2, in which both origins of replication are present. Using NSCV1 and NSCV2, here we tested whether two types of origins of replication can function simultaneously on the same chromosome or one or the other origin is silenced. We found that in NSCV1, both origins are active whereas in NSCV2  ori2  is silenced despite the fact that it is functional in an isolated context. The  ori2  activity appears to be primarily determined by the copy number of the triggering site,  crtS,  which in turn is determined by its location with respect to  ori1  and  ori2  on the fused chromosome.

Functionality of Two Origins of Replication in Vibrio cholerae Strains With a Single Chromosome