Fig. 4: Physical interaction between WspR with RpfG. | npj Biofilms and Microbiomes

Fig. 4: Physical interaction between WspR with RpfG.

From: The Wsp chemosensory system modulates c-di-GMP-dependent biofilm formation by integrating DSF quorum sensing through the WspR-RpfG complex in Lysobacter

Fig. 4

a E. coli-based B2H assay showing that WspR interacts with RpfG. CK+, positive control (pBT-GacS and pTRG-GacS) and CK−, negative control (empty vector of pBT and pTRG). WspR, pTRG-WspR. RpfG, pBT-RpfG. −3AT-Strr and +3AT + Strr represent strains grown on LB-based non-selective medium and minimal medium (M9)-based selective medium respectively. b Pull-down assay confirming the interaction of WspR-His with RpfG-FLAG. The IP assay was performed using anti-FLAG antibody. Western blotting was carried out using anti-FLAG and anti-His antibodies. c Microscale thermophoresis (MST) measurement of binding affinity: green trace, WspR-His and MBP-RpfG (Kd = 0.15 μM); red trace, WspR-His and MBP (no binding). d Schematic map of the full-length WspR and their truncations (WspRΔREC and WspRΔEEF). e Schematic map of the full-length RpfG and their truncations (RpfGΔREC and RpfGΔGYP). f Pull-down assay confirming the interactions between the GGDEF domains of WspR (WspRΔREC) and the HD-GYP domain of RpfG (RpfGΔREC). All blots derive from the same experiment and that they were processed in parallel.

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