Effects of recJ, recQ, and recFOR mutations on recombination in nuclease-deficient recB recD double mutants of Escherichia coli.
The two main recombination pathways in Escherichia coli (RecBCD and RecF) have different recombination machineries that act independently in the initiation of recombination. Three essential enzymatic activities are required for early recombinational processing of double-stranded DNA ends and breaks: a helicase, a 5'-->3' exonuclease, and loading of RecA protein onto single-stranded DNA tails. The RecBCD enzyme performs all of these activities, whereas the recombination machinery of the RecF pathway consists of RecQ (helicase), RecJ (5'-->3' exonuclease), and RecFOR (RecA-single-stranded DNA filament formation). The recombination pathway operating in recB (nuclease-deficient) mutants is a hybrid because it includes elements of both the RecBCD and RecF recombination machineries. In this study, genetic analysis of recombination in a recB (nuclease-deficient) recD double mutant was performed. We show that conjugational recombination and DNA repair after UV and gamma irradiation in this mutant are highly dependent on recJ, partially dependent on recFOR, and independent of recQ. These results suggest that the recombination pathway operating in a nuclease-deficient recB recD double mutant is also a hybrid. We propose that the helicase and RecA loading activities belong to the RecBCD recombination machinery, while the RecJ-mediated 5'-->3' exonuclease is an element of the RecF recombination machinery.
Ivancic-Bace
Ivana
I
Department of Molecular Biology, Ruder Bosković Institute, Bijenicka 54, HR-10002 Zagreb, Croatia.
Salaj-Smic
Erika
E
Brcic-Kostic
Krunoslav
K
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D003227:Conjugation, Genetic; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D004260:DNA Repair; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000502:physiology; D043211:Exodeoxyribonuclease V / Q000235:genetics / Q000502:physiology; D005090:Exodeoxyribonucleases / Q000235:genetics / Q000502:physiology; D005092:Exonucleases / Q000235:genetics / Q000502:physiology; D005798:Genes, Bacterial; D009154:Mutation; D011995:Recombination, Genetic
D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C081006:RecO protein, E coli; C489318:RecR protein, E coli; C489340:recF protein, E coli; D005090:Exodeoxyribonucleases; D005092:Exonucleases; C489467:RecJ protein, E coli; D043211:Exodeoxyribonuclease V; D000251:Adenosine Triphosphatases; D004265:DNA Helicases