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Type II restriction-modification systems are ubiquitous in prokaryotes. Some of them are present in naturally occurring plasmids, which may facilitate the spread of these systems in bacterial populations by horizontal gene transfer. However, little is known about the routes of their dissemination. As a model to study this, we have chosen an  Escherichia coli  natural plasmid pEC156 that carries the EcoVIII restriction modification system. The presence of this system as well as the  cis -acting  cer  site involved in resolution of plasmid multimers determines the stable maintenance of pEC156 not only in  Escherichia coli  but also in other enterobacteria. We have shown that due to the presence of  oriT -type F and  oriT -type R64 loci it is possible to mobilize pEC156 by conjugative plasmids (F and R64, respectively). The highest mobilization frequency was observed when pEC156-derivatives were transferred between  Escherichia coli  strains,  Enterobacter cloacae  and  Citrobacter freundii  representing coliform bacteria. We found that a pEC156-derivative with a functional EcoVIII restriction-modification system was mobilized in enterobacteria at a frequency lower than a plasmid lacking this system. In addition, we found that bacteria that possess the EcoVIII restriction-modification system can efficiently release plasmid content to the environment. We have shown that  E .  coli  cells can be naturally transformed with pEC156-derivatives, however, with low efficiency. The transformation protocol employed neither involved chemical agents (e.g. CaCl 2 ) nor temperature shift which could induce plasmid DNA uptake.

plos one

Plasmid pEC156, a Naturally Occurring Escherichia coli Genetic Element That Carries Genes of the EcoVIII Restriction-Modification System, Is Mobilizable among Enterobacteria