FIG 8.
Electrophoretic mobility shift assays for analysis of DNA-binding activity of the Cjj1483 RR. For panels A and B, purified WT Cjj1483 protein was added to various promoter DNAs at concentrations ranging from 0 to 2 μM. For panels C and D, WT Cjj1483 or the Cjj1483D58E or Cjj1483D58N mutant protein was added at concentrations ranging from 0 to 4 μM. For panels A, B, and D, “−AcP” and “+AcP” indicate whether or not Cjj1483 was pretreated with Li-AcP to autophosphorylate the protein prior to addition to DNA-binding assay mixtures. (A) Binding of Cjj1483 to the radiolabeled promoter for Cjj0438 or aphA-3 in the absence of competition. (B) Binding of Cjj1483 to the radiolabeled promoter for Cjj0438 in the presence of unlabeled competitor DNA (Cjj0438 promoter; left panels) or unlabeled noncompetitor DNA (aphA-3 promoter; right panels). (C) Binding of Cjj1483D58E and Cjj1483D58N mutant proteins to the radiolabeled promoter for Cjj0438 in the absence of competition. (D) Binding of Cjj1483 to the radiolabeled promoter for Cjj1386 in the absence of competition.