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. 2021 Aug 3;12:706934. doi: 10.3389/fmicb.2021.706934

FIGURE 4.

FIGURE 4

YmoA-mediated control of CsrC occurs on the post-transcriptional level. (A) Model for YmoA influence on CsrC on the transcriptional or post-transcriptional level. Increased CsrC levels can result from increased transcription or decreased degradation of CsrC. (B) Plasmids harboring transcriptional csrC-lacZ fusions with varying 3′-ends of the csrC gene and a csrC(+81)-lacZ fusion with a Δ(+24–+57) deletion (pKB17) were transformed into Y. pseudotuberculosis YPIII and YP50 (ΔymoA). Strains were grown overnight at 25°C and transcriptional activity of the different fusions was determined by β-galactosidase assays. β-galactosidase activity is given in μmol min–1 mg–1 for comparison. The data represent the average ± SD from at least three different experiments each done in duplicate. The statistical significances between the wild-type and the ymoA mutant were determined by the ANOVA test. P-values: ∗∗∗: <0.001. (C) Secondary structure prediction of the CsrC RNA from nucleotide +1 to +76 generated by Mfold. The single-stranded regions including the most highly conserved GGA element of CsrA binding sites are indicated by numbers. The arrows indicate the 5′ and 3′ end of the Δ(+24–+57) deletion. (D) Y. pseudotuberculosis strains YPIII (wild-type), YP50 (ΔymoA), and YP53 (ΔcsrA) harboring pKB59 (csrC+) or pKB49 (csrCΔ+24–+57) were grown in LB at 25°C overnight. Total RNA of the strains was extracted, separated on agarose gels and a CsrC-specific probe was used to detect the CsrC RNA by Northern blotting. The 16S rRNAs are shown as RNA loading control.