Fig. 3: The promoter of plzR is induced by the second messenger cyclic di-GMP.
From: PlzR regulates type IV pili assembly in Pseudomonas aeruginosa via PilZ binding
A Graphic representation of tested intergenic regions (marked in gray) upstream of genes plzR and cdrA. The predicted promoter of plzR (at position −36 to −81) is marked in black. B Fluorimetric assay to determine the promoter activity of the plzR promoter in the presence of elevated cellular c-di-GMP levels. P. aeruginosa PAO1 strains harboring the yip gene (encoding a phage-encoded diguanylate cyclase YfiN interacting peptide, which induces an increase of c-di-GMP production in the host cell) in the mini-tn7T transposon (PAO1::yip), or the empty counterpart (PAO1::E), supplemented with a reporter plasmid containing the tested promoter region fused to a msfGFP reporter gene were studied in the absence (non-induced) and presence of 1 mM IPTG (induced). As a control, the intergenic region upstream the plzR gene lacking the predicted promoter (pPlzRΔ45bp-msfGFP) was tested. Fluorescent measurements were normalized for OD600nm and corrected for background fluorescent levels of the corresponding strain with empty pSEVE421-plasmid. Data are presented as mean values ± SD of four biological replicates, and p values were calculated using the Student’s t-test (two-sided, n = 4 colonies). Source data are provided as a Source Data file.
