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Listeria monocytogenes  is a food-associated bacterium that is responsible for food-related illnesses worldwide. In the  L. monocytogenes  EGD-e genome, FlhB, FliM, and FliY  (encoded  by  lmo0679, lmo0699 , and  lmo0700 , respectively) are annotated as putative flagella biosynthesis factors, but their functions remain unknown. To explore whether FlhB, FliM, and FliY are involved in  Listeria  flagella synthesis, we constructed  flhB, fliM, fliY , and other flagellar-related gene deletion mutants using a homologous recombination strategy. Then, we analyzed the motility, flagella synthesis, and protein expression of these mutant strains. Motility and flagella synthesis were completely abolished in the absence of  flhB, fliM , or  fliY . These impaired phenotypes were fully restored in the complemented strains CΔ flhB , CΔ fliM , and CΔ fliY . The transcriptional levels of flagellar-related genes, including  flaA, fliM, fliY, lmo0695, lmo0698, fliI , and  fliS , were downregulated markedly in the absence of  flhB, fliM , or  fliY . Deletion of  flhB  resulted in the complete abolishment of FlaA expression, while it decreased FliM and FliY expression. The expression of FlaA was abolished completely in the absence of  fliM  or  fliY . No significant changes were found in the expression of FlhF and two flagella synthesis regulatory factors, MogR and GmaR. We demonstrate for the first time that FlhB, FliM, and FliY not only mediate  Listeria  motility, but also are involved in regulating flagella synthesis. This study provides novel insights that increase our understanding of the roles played by FlhB, FliM, and FliY in the flagellar type III secretion system in  L. monocytogenes .

Flagellar Basal Body Structural Proteins FlhB, FliM, and FliY Are Required for Flagellar-Associated Protein Expression in Listeria monocytogenes