Figure 6: Charged residues within the ClpV1 groove and the TssC1 peptide mediate the ClpV1-TssC1 interaction.

(A) Bacterial two-hybrid assay. BTH101 reporter cells producing the indicated wild-type or mutated proteins fused to the T18 or T25 domains of the Bordetella adenylate cyclase were spotted on X-Gal indicator plates. The blue color of the colony reflects the interaction between the two proteins. (B,C) Sci-1-dependent antibacterial growth inhibition. Prey cells (W3110 gfp⁺, kan^R) were mixed with the indicated attacker cells, spotted onto sci-1-inducing medium (SIM) agar plates and incubated for 4 hours at 37 °C. The image of a representative bacterial spot is shown and the relative fluorescent levels (in arbitrary units, AU) are indicated in the upper graph. The number of recovered E. coli prey cells is indicated in the lower graph (in log₁₀ of colony-forming units (cfu)). The circles indicate values from three independent assays, and the average is indicated by the bar.