The YfiBNR Signal Transduction Mechanism Reveals Novel Targets for the Evolution of Persistent Pseudomonas aeruginosa in Cystic Fibrosis Airways
Figure 5
Effect of YfiB linker mutants.
A) Sequence of the N-terminal YfiB ‘linker’, between the outer membrane (lipid anchor on the highlighted cysteine) and the Pal-like domain. The first activating mutant position (Figure 4) is shown in red. ‘short-linker’ indicates a deletion of 5 amino acids, leaving the lipobox intact. ‘long-linker’ indicates an insertion/duplication of 9 amino acids (dashed line). B) Co-localization of YfiB and YfiR at the outer membrane. Immunoblots of fractionated soluble and membrane samples with anti-YfiB and M2 antisera as shown. ‘YfiB WT’ indicates ΔyfiBNR Tn7::yfiNR containing pME6032-yfiB. ‘short’/‘long’ indicate the strains containing pME6032-yfiB linker mutants. C) The effect of different yfiB mutants, expressed from pME6032 in ΔyfiBNR Tn7::yfiNR, on attachment is shown relative to pME6032 only (ctrl.). The immunoblot shows the levels of YfiB protein present in each strain. D) Colony morphologies on LB Congo-red agar upon over-expression of yfiB mutants.