title: Directed evolution of ionizing radiation resistance in Escherichia coli.
abstract: We have generated extreme ionizing radiation resistance in a relatively sensitive bacterial species, Escherichia coli, by directed evolution. Four populations of Escherichia coli K-12 were derived independently from strain MG1655, with each specifically adapted to survive exposure to high doses of ionizing radiation. D(37) values for strains isolated from two of the populations approached that exhibited by Deinococcus radiodurans. Complete genomic sequencing was carried out on nine purified strains derived from these populations. Clear mutational patterns were observed that both pointed to key underlying mechanisms and guided further characterization of the strains. In these evolved populations, passive genomic protection is not in evidence. Instead, enhanced recombinational DNA repair makes a prominent but probably not exclusive contribution to genome reconstitution. Multiple genes, multiple alleles of some genes, multiple mechanisms, and multiple evolutionary pathways all play a role in the evolutionary acquisition of extreme radiation resistance. Several mutations in the recA gene and a deletion of the e14 prophage both demonstrably contribute to and partially explain the new phenotype. Mutations in additional components of the bacterial recombinational repair system and the replication restart primosome are also prominent, as are mutations in genes involved in cell division, protein turnover, and glutamate transport. At least some evolutionary pathways to extreme radiation resistance are constrained by the temporally ordered appearance of specific alleles.
authors.affiliation: Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53575, USA.
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Harris
authors.forename: Steve V
authors.initials: SV
authors.lastname: Pollock
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Wood
authors.forename: Reece J
authors.initials: RJ
authors.lastname: Goiffon
authors.forename: Audrey J
authors.initials: AJ
authors.lastname: Klingele
authors.forename: Eric L
authors.initials: EL
authors.lastname: Cabot
authors.forename: Wendy
authors.initials: W
authors.lastname: Schackwitz
authors.forename: Joel
authors.initials: J
authors.lastname: Martin
authors.forename: Julie
authors.initials: J
authors.lastname: Eggington
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Durfee
authors.forename: Christina M
authors.initials: CM
authors.lastname: Middle
authors.forename: Jason E
authors.initials: JE
authors.lastname: Norton
authors.forename: Michael C
authors.initials: MC
authors.lastname: Popelars
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.forename: Sarit A
authors.initials: SA
authors.lastname: Klugman
authors.forename: Lindsay L
authors.initials: LL
authors.lastname: Hamilton
authors.forename: Lukas B
authors.initials: LB
authors.lastname: Bane
authors.forename: Len A
authors.initials: LA
authors.lastname: Pennacchio
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Albert
authors.forename: Nicole T
authors.initials: NT
authors.lastname: Perna
authors.forename: Michael M
authors.initials: MM
authors.lastname: Cox
authors.forename: John R
authors.initials: JR
authors.lastname: Battista
mesh_terms: D002851:Chromatography, High Pressure Liquid; D019020:Directed Molecular Evolution; D016521:Electrophoresis, Gel, Pulsed-Field; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects*; D009154:Mutation; D010802:Phylogeny; D011839:Radiation, Ionizing; D011938:Rec A Recombinases / Q000235:genetics / Q000502:physiology
chemical_list: D011938:Rec A Recombinases