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. 2017 May 23;85(6):e00121-17. doi: 10.1128/IAI.00121-17

FIG 6.

FIG 6

The PTSNtr affects the levels of expression of PmrA-regulated genes via PmrA. (A) The expression of the PmrA translational lacZ fusion was examined in the wild-type strain (JR32), in the ptsP deletion mutant (SY-ΔptsP), in the ptsO deletion mutant (SY-ptsO-Km), and in the ptsP-ptsO double deletion mutant (SY-ΔptsP-ptsO-Km) at the stationary phase. (B) The expression of effector translational lacZ fusions (the effectors examined are indicated below the bars) was examined in the wild-type strain (JR32), in the ptsP deletion mutant (SY-ΔptsP), in the pmrA deletion mutant (HK-PQ1), and in the ptsP-pmrA double-deletion mutant (SY-ΔptsP-pmrA-Km) at the stationary phase. The levels of expression of the marked lacZ fusions were found to be significantly different (*, P < 0.01; **, P < 0.001, Student's t test). For panels A and B, strains are identified according to the legend on the figure. (C) The levels of expression of two PmrA-regulated effector-encoding genes (ceg3 and lepB) were examined in the L. pneumophila wild-type strain (JR32) and the pmrA deletion mutant (HK-PQ1). The bacteria contained a plasmid with the ptsP gene (two upper graphs) or the ptsO gene (two lower graphs) cloned under the control of the IPTG-inducible Ptac promoter. The plasmids containing the corresponding lacZ fusion of the examined gene without the ptsP or ptsO gene were used as controls (white bars). The IPTG concentrations used to express the wild-type and mutated ptsP were 0.005, 0.01, and 0.05 mM, and they were 0, 0.005, and 0.01 mM for ptsO. The levels of expression of the lacZ fusions were found to be significantly different (*, P < 0.01; **, P < 0.001, Student's t test), in comparisons between results with the fusions with the ptsP or ptsO gene expressed from the Ptac promoter and those with the corresponding lacZ fusions without the ptsP and ptsO genes. β-Galactosidase activity was measured as described in Materials and Methods. Data (expressed in Miller units [M.U.]) are the averages ± standard deviations (error bars) of the results of at least three different experiments.