The FlhA linker mediates flagellar protein export switching during flagellar assembly

The flagellar protein export apparatus switches substrate specificity from hook-type to filament-type upon hook assembly completion, thereby initiating filament assembly at the hook tip. The C-terminal cytoplasmic domain of FlhA (FlhA_C) serves as a docking platform for flagellar chaperones in complex with their cognate filament-type substrates. Interactions of the flexible linker of FlhA (FlhA_L) with its nearest FlhA_C subunit in the FlhA_C ring is required for the substrate specificity switching. To address how FlhA_L brings the order to flagellar assembly, we analyzed the flhA(E351A/W354A/D356A) ΔflgM mutant and found that this triple mutation in FlhA_L increased the secretion level of hook protein by 5-fold, thereby increasing hook length. The crystal structure of FlhA_C(E351A/D356A) showed that FlhA_L bound to the chaperone-binding site of its neighboring subunit. We propose that the interaction of FlhA_L with the chaperon-binding site of FlhA_C suppresses filament-type protein export and facilitates hook-type protein export during hook assembly.