title: Highly variable individual donor cell fates characterize robust horizontal gene transfer of an integrative and conjugative element.
abstract: Horizontal gene transfer is an important evolutionary mechanism for bacterial adaptation. However, given the typical low transfer frequencies in a bacterial population, little is known about the fate and interplay of donor cells and the mobilized DNA during transfer. Here we study transfer of an integrative and conjugative element (ICE) among individual live bacterial cells. ICEs are widely distributed mobile DNA elements that are different than plasmids because they reside silent in the host chromosome and are maintained through vertical descent. Occasionally, ICEs become active, excise, and transmit their DNA to a new recipient, where it is reintegrated. We develop a fluorescent tool to differentiate excision, transfer, and reintegration of a model ICE named ICEclc (for carrying the clc genes for chlorocatechol metabolism) among single Pseudomonas cells by using time-lapse microscopy. We find that ICEclc activation is initiated in stationary phase cells, but excision and transfer predominantly occur only when such cells have been presented with new nutrients. Donors with activated ICE develop a number of different states, characterized by reduced cell division rates or growth arrest, persistence, or lysis, concomitant with ICE excision, and likely, ICE loss or replication. The donor cell state transitions can be described by using a stochastic model, which predicts that ICE fitness is optimal at low initiation rates in stationary phase. Despite highly variable donor cell fates, ICE transfer is remarkably robust overall, with 75% success after excision. Our results help to better understand ICE behavior and shed a new light on bacterial cellular differentiation during horizontal gene transfer.
authors.affiliation: Department of Fundamental Microbiology, Bâtiment Biophore, University of Lausanne, 1015 Lausanne, Switzerland.
authors.forename: François
authors.initials: F
authors.lastname: Delavat
authors.affiliation: Department of Fundamental Microbiology, Bâtiment Biophore, University of Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Sara
authors.initials: S
authors.lastname: Mitri
authors.affiliation: Department of Fundamental Microbiology, Bâtiment Biophore, University of Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Serge
authors.initials: S
authors.lastname: Pelet
authors.affiliation: Department of Fundamental Microbiology, Bâtiment Biophore, University of Lausanne, 1015 Lausanne, Switzerland janroelof.vandermeer@unil.ch.
authors.forename: Jan Roelof
authors.initials: JR
authors.lastname: van der Meer
mesh_terms: D002455:Cell Division / Q000502:physiology*; D003227:Conjugation, Genetic / Q000502:physiology*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D022761:Gene Transfer, Horizontal / Q000502:physiology*; D008954:Models, Biological; D016958:Pseudomonas putida / Q000235:genetics / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial
keywords: Pseudomonas; bacterial evolution; conjugation; modeling; time-lapse microscopy