Rapid Evolution of Citrate Utilization by Escherichia coli by Direct Selection Requires citT and dctA

Journal of Bacteriology
Dustin J Van HofwegenScott A Minnich

Abstract

The isolation of aerobic citrate-utilizing Escherichia coli (Cit(+)) in long-term evolution experiments (LTEE) has been termed a rare, innovative, presumptive speciation event. We hypothesized that direct selection would rapidly yield the same class of E. coli Cit(+) mutants and follow the same genetic trajectory: potentiation, actualization, and refinement. This hypothesis was tested with wild-type E. coli strain B and with K-12 and three K-12 derivatives: an E. coli ΔrpoS::kan mutant (impaired for stationary-phase survival), an E. coli ΔcitT::kan mutant (deleted for the anaerobic citrate/succinate antiporter), and an E. coli ΔdctA::kan mutant (deleted for the aerobic succinate transporter). E. coli underwent adaptation to aerobic citrate metabolism that was readily and repeatedly achieved using minimal medium supplemented with citrate (M9C), M9C with 0.005% glycerol, or M9C with 0.0025% glucose. Forty-six independent E. coli Cit(+) mutants were isolated from all E. coli derivatives except the E. coli ΔcitT::kan mutant. Potentiation/actualization mutations occurred within as few as 12 generations, and refinement mutations occurred within 100 generations. Citrate utilization was confirmed using Simmons, Christensen, and LeMaste...Continue Reading

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Citations

Mar 23, 2019·Annals of the New York Academy of Sciences·James A Shapiro
Jun 20, 2018·Proceedings of the National Academy of Sciences of the United States of America·Melissa Tumen-VelasquezEllen L Neidle
Jul 27, 2018·Microbiology and Molecular Biology Reviews : MMBR·Bram Van den BerghJan Michiels
Nov 10, 2018·Science·Zachary D BlountJonathan B Losos
Nov 19, 2019·Frontiers in Genetics·Christine J YeHenry H Heng
Feb 18, 2018·Research in Microbiology·Lacey L WestphalKarin E Kram
May 7, 2021·ACS Sensors·Lauren M CastleAriel L Furst
May 13, 2021·Progress in Biophysics and Molecular Biology·Keith Baverstock
Sep 10, 2021·PLoS Pathogens·Matthew J Culyba, Daria Van Tyne

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