Directed evolution of ionizing radiation resistance in Escherichia coli.
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.
Harris
Dennis R
DR
Department of Biochemistry, University of Wisconsin-Madison, 433 Babcock Drive, Madison, WI 53575, USA.
Pollock
Steve V
SV
Wood
Elizabeth A
EA
Goiffon
Reece J
RJ
Klingele
Audrey J
AJ
Cabot
Eric L
EL
Schackwitz
Wendy
W
Martin
Joel
J
Eggington
Julie
J
Durfee
Timothy J
TJ
Middle
Christina M
CM
Norton
Jason E
JE
Popelars
Michael C
MC
Li
Hao
H
Klugman
Sarit A
SA
Hamilton
Lindsay L
LL
Bane
Lukas B
LB
Pennacchio
Len A
LA
Albert
Thomas J
TJ
Perna
Nicole T
NT
Cox
Michael M
MM
Battista
John R
JR
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
D011938:Rec A Recombinases