title: Rep and PriA helicase activities prevent RecA from provoking unnecessary recombination during replication fork repair.
abstract: The rescue of replication forks stalled on the template DNA was investigated using an assay for synthetic lethality that provides a visual readout of cell viability and permits investigation of why certain mutations are lethal when combined. The results presented show that RecA and other recombination proteins are often engaged during replication because RecA is present and provokes recombination rather than because recombination is necessary. This occurs particularly frequently in cells lacking the helicase activities of Rep and PriA. We propose that these two proteins normally limit the loading of RecA on ssDNA regions exposed on the leading strand template of damaged forks, and do so by unwinding the nascent lagging strand, thus facilitating reannealing of the parental strands. Gap closure followed by loading of the DnaB replicative helicase enables synthesis of the leading strand to continue. Without either activity, RecA loads more frequently on the DNA and drives fork reversal, which creates a chickenfoot structure and a requirement for other recombination proteins to re-establish a viable fork. The assay also reveals that stalled transcription complexes are common impediments to fork progression, and that damaged forks often reverse independently of RecA.
authors.affiliation: Institute of Genetics, University of Nottingham, Queen's Medical Centre, UK.
authors.forename: Akeel A
authors.initials: AA
authors.lastname: Mahdi
authors.forename: Carol
authors.initials: C
authors.lastname: Buckman
authors.forename: Lynda
authors.initials: L
authors.lastname: Harris
authors.forename: Robert G
authors.initials: RG
authors.lastname: Lloyd
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D004261:DNA Replication; D004269:DNA, Bacterial; D011938:Rec A Recombinases / Q000235:genetics / Q000378:metabolism*; D011995:Recombination, Genetic
chemical_list: D004269:DNA, Bacterial; D011938:Rec A Recombinases; D000251:Adenosine Triphosphatases; D004265:DNA Helicases