Specificity of motor components in the dual flagellar system of Shewanella putrefaciens CN-32

Summary Bacterial flagellar motors are intricate nanomachines in which the stator units and rotor component FliM may be dynamically exchanged during function. Similar to other bacterial species, the gammaproteo- bacterium Shewanella putrefaciens CN-32 possesses a complete secondary flagellar system along with a corresponding stator unit. Expression of the secondary system occurs during planktonic growth in complex media and leads to the formation of a subpopulation with one or more additional flagella at random positions in addition to the primary polar system. We used physiological and phenotypic char- acterizations of defined mutants in concert with fluo- rescent microscopy on labelled components of the two different systems, the stator proteins PomB and MotB, the rotor components FliM1 and FliM2, and the auxiliary motor components MotX and MotY, to deter- mine localization, function and dynamics of the pro- teins in the flagellar motors. The results demonstrate that the polar flagellum is driven by a Na + -dependent FliM1/PomAB/MotX/MotY flagellar motor while the secondary system is rotated by a H + -dependent FliM2/MotAB motor. The components were highly spe- cific for their corresponding motor and are unlikely to be extensively swapped or shared between the two flagellar systems under planktonic conditions. The results have implications for both specificity and dynamics of flagellar motor components.