Antagonists of DNA gyrase inhibit repair and recombination of UV-irradiated phage lambda

Abstract

Intracellular lambda DNA (from EDTA-sensitive tandem duplication phages) was extracted from infected rec+ bacteria and scored for infectivity and recombination (loss of duplication) by transfection of recA recB spheroplasts and subsequent assay for EDTA resistance. When phage development was blocked by repressor or by antibiotics (chloramphenicol and/or rifampin), the apparent recombination frequency was about 0.1% above the background value for recA infections. Prior irradiation of the phage greatly stimulated recombination; the frequency was 20% when UV fluence was 140 J/m2. Repair (recovery of infectivity) and recombination of irradiated phage DNA proceeded readily in the presence of chloramphenicol and rifampin. Inhibitors of DNA gyrase (coumermycin and oxolinic acid) blocked repair and reduced recombination. UV-stimulated recombination was very low in recA but nearly normal in recB cells: repair was reduced in both mutant strains. The recombination remained high as phage/cell ratios less than unity.