title: recA730-dependent suppression of recombination deficiency in RecA loading mutants of Escherichia coli.
abstract: Homologous recombination is an essential process in double-strand break repair. The main requirement for recombination is formation of a RecA filament. Double-strand breaks can be processed into a RecA filament by the action of three enzymatic activities: helicase, 5'-3' exonuclease and RecA loading onto ssDNA. These activities are provided by the RecBCD enzyme in wild type cells or by the RecF pathway gene products in recBC sbcBC(D) cells. In the recBD1080A mutant (recB∗ mutant), the recombination machineries of RecBCD and RecF pathways are interchangeable and include RecB∗CD enzyme (helicase), RecJ (5'-3' exonuclease) and RecFOR (RecA loading). The mutant RecA730 protein is able to produce a RecA filament without the help of RecFOR mediators, since it more efficiently competes with SSB protein for ssDNA than the normal RecA protein. It was previously shown that the recA730 mutation suppresses UV sensitivity in a uvrA recFOR genetic background. We tested whether the recA730 mutation can suppress recombination and DNA repair deficiency in a recB∗ mutant and its derivatives. We show that the recA730 mutation suppresses recombination deficiency in a recB∗ recFOR background, where the defect is at the level of RecA loading, but not in the recB∗ recJ background where the defect is at the level of nuclease activity.
authors.affiliation: Department of Molecular Biology, Ruđer Bošković Institute, Bijenička 54, 10000 Zagreb, Croatia.
authors.forename: Ignacija
authors.initials: I
authors.lastname: Vlašić
authors.forename: Ana
authors.initials: A
authors.lastname: Simatović
authors.forename: Krunoslav
authors.initials: K
authors.lastname: Brčić-Kostić
mesh_terms: D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism*; D043211:Exodeoxyribonuclease V / Q000235:genetics; D017353:Gene Deletion; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D011938:Rec A Recombinases / Q000235:genetics* / Q000378:metabolism*; D011995:Recombination, Genetic; D013489:Suppression, Genetic
chemical_list: D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D050505:Mutant Proteins; C489340:recF protein, E coli; D011938:Rec A Recombinases; D043211:Exodeoxyribonuclease V