Modulation of recombination and DNA repair by the RecG and PriA helicases of Escherichia coli K-12.
The RecG protein of Escherichia coli is a structure-specific DNA helicase that targets strand exchange intermediates in genetic recombination and drives their branch migration along the DNA. Strains carrying null mutations in recG show reduced recombination and DNA repair. Suppressors of this phenotype, called srgA, were located close to metB and shown to be alleles of priA. Suppression depends on the RecA, RecBCD, RecF, RuvAB, and RuvC recombination proteins. Nine srgA mutations were sequenced and shown to specify mutant PriA proteins with single amino acid substitutions located in or close to one of the conserved helicase motifs. The mutant proteins retain the ability to catalyze primosome assembly, as judged by the viability of recG srgA and srgA strains and their ability to support replication of plasmids based on the ColE1 replicon. Multicopy priA+ plasmids increase substantially the recombination- and repair-deficient phenotype of recG strains and confer similar phenotypes on recG srgA double mutants but not on ruvAB or wild-type strains. The multicopy effect is eliminated by K230R, C446G, and C477G substitutions in PriA. It is concluded that the 3'-5' DNA helicase/translocase activity of PriA inhibits recombination and that this effect is normally countered by RecG.
Al-Deib
A A
AA
Department of Genetics, University of Nottingham, Queens Medical Centre, United Kingdom.
Mahdi
A A
AA
Lloyd
R G
RG
D000251:Adenosine Triphosphatases / Q000235:genetics / Q000378:metabolism*; D000483:Alleles; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003227:Conjugation, Genetic; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000528:radiation effects; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D016153:Genes, Suppressor; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D011995:Recombination, Genetic; D013489:Suppression, Genetic; D014466:Ultraviolet Rays
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C071446:RecG protein, E coli; D000251:Adenosine Triphosphatases; C506743:priA protein, E coli; D004265:DNA Helicases