Fig. 5: Interaction of MrpC and kinase complexes at the promoter region of mrpC.

a Interaction of AskA, AskB, AskC and MrpC with PmrpC-DNA as determined by DNA-protein pull-down assays. Scans of representative SDS-PAGE gels resolving proteins captured in various DNA-protein pull-down experiments are shown. Protein bands are marked and named exemplarily. Biotinylated PmrpC-DNA (probe), ambruticin VS-3, as well as purified proteins AskA, AskB, AskC, and MrpC were mixed in different combinations in the presence of ATP. Proteins, which bound to the biotinylated PmrpC-DNA, were extracted from the mixtures by using streptavidin-coated magnetic beads. The composition of each attempt is indicated below the lanes. Lanes 1 and 8: PageRuler plus prestained protein ladder (Thermo); Lanes 12, 16, and 19: PageRuler prestained protein ladder (Thermo). b Interaction of AskA, AskB, AskC and MrpC as determined by protein-protein pull-down assays. Scans of representative SDS-PAGE gels resolving proteins captured in various protein-protein pull-down experiments are shown. Protein bands are marked and named exemplarily. The protein band marked by a white arrow is an N-terminal fragment of AskB, which could not be separated from full-length AskB upon purification. Other minor impurities are all smaller N-terminal fragments of AskA or AskB, which also could not be separated. C-terminally His6-tagged AskC (probe), ambruticin VS-3, as well as purified proteins AskA, AskB and MrpC were mixed in different combinations. Proteins, which bound to the C-terminally-His’s-tagged AskC, were extracted from the mixtures by using Dynabeads® His-Tag Isolation & Pulldown magnetic beads. The composition of each attempt is indicated below the lanes. Lanes 1 and 15: PageRuler prestained protein ladder (Thermo); Lanes 8 and 16: PageRuler plus prestained protein ladder (Thermo). Results shown in panels a and b were performed in duplicates. Source data are provided as a Source Data file.