PriA supports two distinct pathways for replication restart in UV-irradiated Escherichia coli cells.
The Escherichia coli PriA protein loads the DnaB replicative helicase at branched DNA structures independently of the replication initiator protein, DnaA, and thereby facilitates assembly of functional replisomes at sites removed from oriC. It is therefore a critical factor in the rescue of replication forks stalled at DNA lesions. It is also a DNA helicase. We describe insertions near the 3' end of priA that interfere with PriA activity. These insertions and the previously described priA300 encoding helicase-defective PriA K230R are shown to be effective suppressors of the DNA repair defect in recG strains, but substantially reduce the ability of ruv mutants to survive DNA damage. The data presented suggest that PriA helicase in conjunction with RecG can promote direct rescue of stalled forks independently of the recombinational pathway promoted by the combined activities of the RuvABC, RecBCD and RecA proteins, which requires only the primosome assembly activity of PriA to load DnaB at D loops. In cells lacking the helicase activity of PriA, we propose that stalled forks can be redirected to the recombination pathway via a Holliday junction intermediate common to both pathways, thus explaining the resistance of these cells to DNA damage.
Jaktaji
Razieh P
RP
Institute of Genetics, University of Nottingham, Queen's Medical Centre, Nottingham NG7 2UH, UK.
Lloyd
Robert G
RG
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D004249:DNA Damage; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects*; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000235:genetics / Q000528:radiation effects; D053488:DnaB Helicases; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects*; D029968:Escherichia coli Proteins; D008954:Models, Biological; D016254:Mutagenesis, Insertional; D011995:Recombination, Genetic; D013489:Suppression, Genetic; D014466:Ultraviolet Rays
D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D000251:Adenosine Triphosphatases; C506730:dnaB protein, E coli; C506743:priA protein, E coli; D004265:DNA Helicases; D053488:DnaB Helicases