DNA damage differentially activates regional chromosomal loci for Tn7 transposition in Escherichia coli.
The bacterial transposon Tn7 recognizes replicating DNA as a target with a preference for the region where DNA replication terminates in the Escherichia coli chromosome. It was previously shown that DNA double-strand breaks in the chromosome stimulate Tn7 transposition where transposition events occur broadly around the point of the DNA break. We show that individual DNA breaks actually activate a series of small regional hotspots in the chromosome for Tn7 insertion. These hotspots are fixed and become active only when a DNA break occurs in the same region of the chromosome. We find that the distribution of insertions around the break is not explained by the exonuclease activity of RecBCD moving the position of the DNA break, and stimulation of Tn7 transposition is not dependent on RecBCD. We show that other forms of DNA damage, like exposure to UV light, mitomycin C, or phleomycin, also stimulate Tn7 transposition. However, inducing the SOS response does not stimulate transposition. Tn7 transposition is not dependent on any known specific pathway of replication fork reactivation as a means of recognizing DNA break repair. Our results are consistent with the idea that Tn7 recognizes DNA replication involved in DNA repair and reveals discrete regions of the chromosome that are differentially activated as transposition targets.
Department of Microbiology, Cornell University, Ithaca, New York 14853, USA.
Qiaojuan
Q
Shi
Adam R
AR
Parks
Benjamin D
BD
Potter
Ilan J
IJ
Safir
Yun
Y
Luo
Brian M
BM
Forster
Joseph E
JE
Peters
D002876:Chromosomes, Bacterial / Q000235:genetics*; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects / Q000528:radiation effects; D004249:DNA Damage; D004265:DNA Helicases / Q000378:metabolism; D004261:DNA Replication / Q000187:drug effects / Q000528:radiation effects; D004251:DNA Transposable Elements / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000378:metabolism; D016685:Mitomycin / Q000494:pharmacology; D008957:Models, Genetic; D010692:Phleomycins / Q000494:pharmacology; D013014:SOS Response, Genetics / Q000187:drug effects / Q000528:radiation effects; D012641:Selection, Genetic; D014466:Ultraviolet Rays
D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D010692:Phleomycins; D016685:Mitomycin; C506743:priA protein, E coli; D004265:DNA Helicases