title: High-copy bacterial plasmids diffuse in the nucleoid-free space, replicate stochastically and are randomly partitioned at cell division.
abstract: Bacterial plasmids play important roles in the metabolism, pathogenesis and bacterial evolution and are highly versatile biotechnological tools. Stable inheritance of plasmids depends on their autonomous replication and efficient partition to daughter cells at cell division. Active partition systems have not been identified for high-copy number plasmids, and it has been generally believed that they are partitioned randomly at cell division. Nevertheless, direct evidence for the cellular location of replicating and nonreplicating plasmids, and the partition mechanism has been lacking. We used as model pJHCMW1, a plasmid isolated from Klebsiella pneumoniae that includes two β-lactamase and two aminoglycoside resistance genes. Here we report that individual ColE1-type plasmid molecules are mobile and tend to be excluded from the nucleoid, mainly localizing at the cell poles but occasionally moving between poles along the long axis of the cell. As a consequence, at the moment of cell division, most plasmid molecules are located at the poles, resulting in efficient random partition to the daughter cells. Complete replication of individual molecules occurred stochastically and independently in the nucleoid-free space throughout the cell cycle, with a constant probability of initiation per plasmid.
authors.affiliation: Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK, Department of Biology, McGill University, Montreal, Quebec H3G 0B1, Canada and Department of Biological Science, Center for Applied Biotechnology Studies, College of Natural Science and Mathematics, California State University Fullerton, Fullerton, CA 92834-6850, USA.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Reyes-Lamothe
authors.forename: Tung
authors.initials: T
authors.lastname: Tran
authors.forename: Diane
authors.initials: D
authors.lastname: Meas
authors.forename: Laura
authors.initials: L
authors.lastname: Lee
authors.forename: Alice M
authors.initials: AM
authors.lastname: Li
authors.forename: David J
authors.initials: DJ
authors.lastname: Sherratt
authors.forename: Marcelo E
authors.initials: ME
authors.lastname: Tolmasky
mesh_terms: D001419:Bacteria / Q000235:genetics*; D002453:Cell Cycle / Q000235:genetics; D002455:Cell Division; D004261:DNA Replication; D004058:Diffusion; D007711:Klebsiella pneumoniae / Q000166:cytology / Q000235:genetics; D010957:Plasmids / Q000096:biosynthesis / Q000502:physiology*; D013269:Stochastic Processes