FliH and FliI help FlhA bring strict order to flagellar protein export in Salmonella

Abstract The flagellar type III secretion system (fT3SS) switches substrate specificity from rod-hook-type to filament-type upon hook completion, terminating hook assembly and initiating filament assembly. The C-terminal cytoplasmic domain of FlhA (FlhA C ) forms a homo-nonameric ring and is directly involved in substrate recognition, allowing the fT3SS to coordinate flagellar protein export with assembly. The highly conserved GYXLI motif (residues 368–372) of FlhA C induces dynamic domain motions of FlhA C required for efficient and robust flagellar protein export by the fT3SS, but it remains unknown whether this motif is also important for ordered protein export by the fT3SS. Here we analyzed two GYXLI mutants, flhA(GAAAA) and flhA(GGGGG) , and provide evidence suggesting that the GYXLI motif in FlhA C requires the flagellar ATPase complex not only to efficiently remodel the FlhA C ring structure for the substrate specificity switching but also to correct substrate recognition errors that occur during flagellar assembly.