Abstract
Campylobacter jejuni is an enteric pathogen and a major cause of foodborne illness worldwide. It has been shown that C. jejuni primarily utilizes amino acids as its preferred energy source, but its ability to utilize L-fucose can grant a competitive advantage during intestinal colonization. In C. jejuni, fucose utilization is encoded by a variable region named plasticity region 2 (PR2); however, the regulatory mechanism for the region remains unknown and is investigated in this study. Genomic sequence analysis revealed that immediately upstream of the fucose utilization operon is a putative IclR-type transcriptional regulator, cj0480c (named fucR here). To determine whether fucR regulates the expression of the fucose utilization operon, we generated a knock-out mutant of fucR. RT-PCR and microarray analysis found that all the genes in the operon were polycistronic and significantly upregulated in the fucR mutant compared with their expression in the wild-type strain. In the presence of fucose, expression of the fucose utilization genes was induced in the wild-type strain but no longer inducible in the fucR mutant, suggesting that FucR functions as a repressor for the fucose utilization operon. To determine whether FucR directly or indirectly regulates the fucose utilization operon, a 6xHis-tagged full-length FucR was produced in Escherichia coli, and the purified recombinant FucR was used in electrophoretic mobility shift assay, which demonstrated that FucR bound specifically to the promoter region of the fucose utilization operon. Together, these results indicate that the L-fucose utilization operon in C. jejuni is directly regulated by FucR, which functions as a transcriptional repressor and modulates the expression of the operon in response to fucose.