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FlhF is a key protein required for complete flagellar synthesis, and its deletion results in the complete absence of a flagella and thus motility in  Campylobacter jejuni . However, the specific mechanism still remains unknown. In this study, RNA-Seq, EMSAs, ChIP-qPCR and β-Galactosidase assays were performed to elucidate the novel interactions between FlhF and genes involved in flagellar synthesis. Results showed that FlhF has an overall influence on the transcription of flagellar genes with an  flhF  mutant displaying down-regulation of most flagellar related genes. FlhF can directly bind to the  flgI  promoter to regulate its expression, which has significant expression change in an  flhF  mutant. The possible binding site of FlhF to the  flgI  promoter was explored by continuously narrowing the  flgI  promoter region and performing further point mutations. Meanwhile, FlhF can directly bind to the promoters of  rpoD ,  flgS , and  fliA  encoding early flagellin regulators, thereby directly or indirectly regulating the synthesis of class I, II, and III flagellar genes, respectively. Collectively, this study demonstrates that FlhF may directly regulate the transcription of flagellar genes by binding to their promoters as a transcriptional regulator, which will be helpful in understanding the mechanism of FlhF in flagellar biosynthetic and bacterial flagellation in general.

Investigating the Role of FlhF Identifies Novel Interactions With Genes Involved in Flagellar Synthesis in Campylobacter jejuni