RecA-independent recombination is efficient but limited by exonucleases.
Genetic recombination in bacteria is facilitated by the RecA strand transfer protein and strongly depends on the homology between interacting sequences. RecA-independent recombination is detectable but occurs at extremely low frequencies and is less responsive to the extent of homology. In this article, we show that RecA-independent recombination in Escherichia coli is depressed by the redundant action of single-strand exonucleases. In the absence of multiple single-strand exonucleases, the efficiency of RecA-independent recombination events, involving either gene conversion or crossing-over, is markedly increased to levels rivaling RecA-dependent events. This finding suggests that RecA-independent recombination is not intrinsically inefficient but is limited by single-strand DNA substrate availability. Crossing-over is inhibited by exonucleases ExoI, ExoVII, ExoX, and RecJ, whereas only ExoI and RecJ abort gene-conversion events. In ExoI(-) RecJ(-) strains, gene conversion can be accomplished by transformation of short single-strand DNA oligonucleotides and is more efficient when the oligonucleotide is complementary to the lagging-strand replication template. We propose that E. coli encodes an unknown mechanism for RecA-independent recombination (independent of prophage recombination systems) that is targeted to replication forks. The potential of RecA-independent recombination to mediate exchange at short homologies suggests that it may contribute significantly to genomic change in bacteria, especially in species with reduced cellular exonuclease activity or those that encode DNA protection factors.
Dutra
Bethany E
BE
Rosenstiel Basic Medical Sciences Research Center and Department of Biology, Brandeis University, Waltham, MA 02454-9110, USA.
Sutera
Vincent A
VA
Lovett
Susan T
ST
D003434:Crossing Over, Genetic; D004260:DNA Repair; D018274:Electroporation; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005090:Exodeoxyribonucleases / Q000502:physiology*; D011938:Rec A Recombinases / Q000502:physiology*; D011995:Recombination, Genetic
D029968:Escherichia coli Proteins; D011938:Rec A Recombinases; D005090:Exodeoxyribonucleases; C489467:RecJ protein, E coli; C028450:exodeoxyribonuclease I; C020133:exodeoxyribonuclease III; C026870:exodeoxyribonuclease VII