Fig. 2

In silico prediction and experimentally engineering of P. putida KT2440 to kill A. hydrophila via T6SS-dependent heterologous effector expression. A Three-dimensional view of the conformational structure of a VgrGkt4 trimer (depicted in blue, red and green) with a monomer of the TplE effector (depicted in purple). B Three-dimensional view of the VgrGkt4-TpleE complex highlighting the residues of TplE involved in the linking with VgrGkt4. Representation was obtained using Alphaphold3 to develop a 3D model and Alphabridge to highlight residues involved in the interaction between proteins. C Alphabridge diagram showing all the interactions between the proteins of the model. Each color represents interactions between different pairs of proteins. D Survival rate of A. hydrophila DSM 30187 (prey) in competition with KT2440 WT or KT2440 Δ3T6SS strains carrying different plasmids (p238, p238-VgrGkt4 and p238-VgrGkt4-TplE) as attackers. For all the experiments, at least three biological replicates are shown, and the error bars represent the SDs. Statistical significance was assessed via one-way ANOVA. **p value < 0.01, *** p value < 0.001, **** p value < 0.0001