Effect and Mechanism Analysis of Pig FUT8 Gene on Resistance to Escherichia coli F18 Infection

Post-weaning diarrhea caused by enterotoxigenic Escherichia coli F18 (E. coli F18) causes significant economic losses for pig producers. Fucosyltransferase 8 (FUT8) is a glycosyltransferase that catalyzes core fucosylation; however, its role in mediating the resistance to E. coli F18 infection in pigs remains unknown. In this study, we systematically verified the relationship between FUT8 expression and E. coli resistance. The results showed that FUT8 was expressed in all detected tissues of Meishan piglets and that its expression was significantly increased in the duodenum and jejunum of E. coli F18-sensitive individuals when compared to E. coli F18-resistant individuals. FUT8 expression increased after exposure to E. coli F18 (p < 0.05) and decreased significantly after LPS induction for 6 h (p < 0.01). Then, the IPEC-J2 stable cell line with FUT8 interference was constructed, and FUT8 knockdown decreased the adhesion of E. coli F18ac to IPEC-J2 cells (p < 0.05). Moreover, we performed a comparative transcriptome study of IPEC-J2 cells after FUT8 knockdown via RNA-seq. In addition, further expression verification demonstrated the significant effect of FUT8 on the glycosphingolipid biosynthesis and Toll-like signaling pathways. Moreover, the core promoter of FUT8, which was located at -1213 bp to -673 bp, was identified via luciferase assay. Interestingly, we found a 1 bp C base insertion mutation at the -774 bp region, which could clearly inhibit the transcriptional binding activity of C/EBP alpha to an FUT8 promoter. Therefore, it is speculated that FUT8 acts in a critical role in the process of E. coli infection; furthermore, the low expression of FUT8 is conducive to the enhancement of E. coli resistance in piglets. Our findings revealed the mechanism of pig FUT8 in regulating E. coli resistance, which provided a theoretical basis for the screening of E. coli resistance in Chinese local pig breeds.

Automated summary

In this study, the relationship between FUT8 expression and E. coli resistance was systematically verified. The results showed that FUT8 was significantly upregulated in the duodenum and jejunum of E. coli F18-sensitive individuals. Knockdown of FUT8 decreased the adhesion of E. coli F18 to cells. Transcriptome analysis revealed the significant effect of FUT8 on glycosphingolipid biosynthesis and Toll-like signaling pathways. The core promoter of FUT8 and a mutation inhibiting its transcriptional binding activity were identified. These findings provide insights into the role of FUT8 in regulating E. coli resistance in pigs.