Fig 3.

Electrophoretic mobility shift assays of PaaR binding to the upstream regions of the paaK, paaA, paaYZ, and paaRI genes. The upstream region of hemH from C. glutamicum AS 1.542 was tested in parallel as a negative reference (A and B). Cy3-labeled DNA probes were added at 0.1 pmol, and PaaR protein was added at 5 pmol (A and D), at 60 pmol (C), or as indicated. The effector PA-CoA was added at 250 μM (A) or as indicated (C). For competition EMSA experiments with the paaRI DNA probe (C) or with the paaK, paaA, or paaYZ DNA probe (D), unlabeled 40-bp DNA fragments carrying the corresponding putative binding sites were added. As a negative control, an unlabeled fragment (40 bp) of the argC gene was added to an EMSA mixture with the Cy3-labeled paaA probe (D).