Extended Data Fig. 5: NER strictly coupled to transcription and is mostly independent of Mfd.
From: Crucial role and mechanism of transcription-coupled DNA repair in bacteria
Supplementary to Fig. 4. a, Wild-type (wt) and mutant cells were UV irradiated at 50 J/m2 and allowed to recover. At the indicated times, genomic DNA was isolated and either treated with T4endoV or mock treated for 30 min at 37 °C and then analysed on alkali-agarose gels. Rifampicin (Rif; 750 µg/ml), chloramphenicol (Cm; 200 µg/ml) and/or bicyclomycin (Bcm; 25 µg/ml) were added 30 min prior to UV irradiation (see Methods). Representative gels are shown for each analysed strain and condition. The percentage of repaired (lesion-free) DNA in T4endoV-treated samples is plotted for each time point relative to untreated samples (see Fig. 4). Data are the mean ± SEM from at least three independent experiments. b, Mfd overexpression interferes with global NER. The wild-type cells containing pMfd and the “empty” vector control (pCA24N) were induced with 0.1 mM IPTG followed by UV irradiation and recovery. The percentage of repaired (lesion-free) DNA in T4endoV-treated samples is plotted for each time point relative to untreated samples. Data are the mean ± SEM from at least three independent experiments.
