Fig. 5: Specific mutations of residues in PilZ and PlzR disturb the PlzR–PilZ–PilB interaction complex.
From: PlzR regulates type IV pili assembly in Pseudomonas aeruginosa via PilZ binding
A Rank 1 ColabFold model of the PlzR–PilZ–PilB interaction complex. The dotted box indicates the region shown magnified at the right. Predicted residues involved in PilZ–PilB interaction (PilZW72 and PilZM118) and in PlzR–PilZ interaction (PilZK43, PilZF52, PlzRD86, and PlzRY90) are indicated, and their sidechains are shown as sticks. B Bacterial two-hybrid results of PilZ and PlzR mutants. PilZ and its mutants were C-terminally fused (indicated with *PilZx) to the T18 domain of the adenylate cyclase, whereas PilB and PlzR and its mutants were C-terminally fused (indicated with *PilB and *PlzRx) to the T25 domain. Combinations with a reduced interaction strength as a result of the mutation are indicated with an asterisk. C Hundredfold serial dilution (100–10−8) of phage LUZ19 dropped on lawns of the P. aeruginosa PAO1 pilZ and plzR double deletion mutant complemented with the IPTG-inducible plzR gene on the mini-T7 transposon and the arabinose-inducible pilZ gene or variants on the pHERD20T plasmid (PAO1ΔplzRΔpilZ::plzR + pilZx). The lipopolysaccharide-dependent phage PEV2 was dropped in parallel for comparison. D Diameter of the twitching zone of the P. aeruginosa PAO1 pilZ and plzR double deletion mutant complemented with the IPTG-inducible plzR gene on the mini-T7 transposon and the arabinose-inducible pilZ gene or variants on the pHERD20T plasmid (PAO1ΔplzRΔpilZ::plzR + pilZx). Data are presented as mean values ± SD and p values were calculated using the Student’s t-test (two-sided, n = 5 biological replicates), n.s. = not significant. Source data are provided as a Source Data file. E Tenfold serial dilution (10−1–10−8) of phage LUZ19 dropped on lawns of the P. aeruginosa PAO1 plzR deletion mutant carrying the plzR gene or a mutant (plzRD86A, plzRY90A) on the mini-tn7T transposon (PAO1ΔplzR::plzRx). The lipopolysaccharide-dependent phage PEV2 was dropped in parallel for comparison. F Diameter of twitching zone of the P. aeruginosa PAO1 plzR deletion mutant complemented with the IPTG-inducible plzR gene or a mutant on the mini-T7 transposon (PAO1ΔplzR::plzRx). Data are presented as mean values ± SD and p values were calculated using the Student’s t-test (two-sided, n = 5 biological replicates). Source data are provided as a Source Data file.
