Regulation of the rulAB Mutagenic DNA Repair Operon of Pseudomonas syringae by UV-B (290 to 320 Nanometers) Radiation and Analysis of rulAB -Mediated Mutability In Vitro and In Planta

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Abstract
The effects of the rulAB operon ofPseudomonas syringae on mutagenic DNA repair and the transcriptional regulation of rulAB following irradiation with UV-B wavelengths were determined. For arulB::Km insertional mutant constructed inP. syringae pv. syringae B86-17, sensitivity to UV-B irradiation increased and UV mutability decreased by 12- to 14-fold.rulAB-induced UV mutability was also tracked in phyllosphere populations of B86-17 for up to 5 days following plant inoculation. UV mutability to rifampin resistance (Rifr) was detected at all sampling points at levels which were significantly greater than in nonirradiated controls. In P. aeruginosa PAO1, the cloned rulABdeterminant on pJJK17 conferred a 30-fold increase in survival and a 200-fold increase in mutability following a UV-B dose of 1,900 J m−2. In comparative studies using defined genetic constructs, we determined that rulAB restored mutability to the Escherichia coli umuDC deletion mutant RW120 at a level between those of its homologs mucAB and umuDC. Analyses using a rulAB::inaZtranscriptional fusion in Pseudomonas fluorescens Pf5 showed that rulAB was rapidly induced after UV-B irradiation, with expression levels peaking at 4 h. At the highest UV-B dose administered, transcriptional activity of therulAB promoter was elevated as much as 261-fold compared to that of a nonirradiated control. The importance of rulABfor survival of P. syringae in its phyllosphere habitat, coupled with its wide distribution among a broad range of P. syringae genotypes, suggests that this determinant would be appropriate for continued investigations into the ecological ramifications of mutagenic DNA repair.