Figure 3.
RepX as new ligand of FinO. (A) Schematic representation of pRSF1010 plasmid present in SL1344. In yellow antibiotic resistance cassetes to streptomycin and sulfonamides. In pink, loci involved in mobilization of the plasmid. In green loci involved in replication of the plasmid. In grey loci with unknown function. RepX in purple antisense to repA locus. (B) Northern blot detection of RepX co-immunoprecipitation with FinO or ProQ. (C) Total RNA levels of FinP and RepX levels in WT, ΔfinO and ΔproQ strains detected by northern blot. 5S was detected as a loading control. (D) Stability of FinP and RepX in WT, ΔfinO and ΔproQ strains determined by detection of FinP and RepX by northern blot after rifampicin treatment. Samples were collected at time points 0, 15, 30, 60, 120 and 240 min. Right panels, quantifications of FinP and RepX stability. (E) Relative plasmid copy number of pRSF1010 in WT and ΔfinO strains. The ratio plasmid pRSF1010/chromosome was quantified by qPCR in WT and ΔfinO extracts from cultures at OD600 nm 2.0. Average of WT values was set to 1 to determine fold change in ΔfinO extracts. Unpaired two-tailed Student's t-test was carried out, ****P < 0.0001.