Abstract
Escherichia coli (E. coli) F18 is the main pathogen responsible for post-weaning diarrhea (PWD) in piglets. Resistance to E. coli F18 depends on the expression of the cognate receptors in the intestinal epithelial cells. However, the molecular mechanism of E. coli F18 resistance in weaned piglets remains unclear. Here, we performed a comparative transcriptome study of the duodenal tissue from Sutai E. coli F18 sensitive and resistant pigs by RNA-seq, and pig α(1,2) fucosyltransferase 2 (FUT2) was identified as a host differentially expressed gene controlling the E. coli F18 infection. Function analysis showed that the FUT2 expression was high in the duodenum and jejunum, with higher levels detected in sensitive individuals than in resistant individuals (p < 0.01). Expression levels of FUT2 were upregulated in IPEC-J2 cells after lipopolysaccharide (LPS)-induction or E. coli stimulation. FUT2 knockdown decreased the adhesion of E. coli F18 to IPEC-J2 cells (p < 0.05). FUT2 overexpression markedly increased the adhesion of E. coli F18 to IPEC-J2 cells (p < 0.05 or p < 0.01). Furthermore, the FUT2 mRNA levels correlated with methylation levels of the mC-22 site in the specificity protein 1 (Sp1) transcription factor (p < 0.05). Electrophoretic mobility shift assays (EMSA) showed that Sp1 interacts with the wild-type FUT2 promoter DNA, but not with methylated DNA. Our data suggested that FUT2 methylation at the mC-22 site inhibits Sp1 binding to the FUT2 promoter, thereby reducing FUT2 expression and enhancing E. coli F18 resistance in weaned piglets. These observations highlight FUT2 as a promising new target for combating E. coli F18 susceptibility in weaned piglets.
Highlights
Porcine post-weaning diarrhea (PWD) causes serious economic losses to large-scale pig farms
We identified the differential expression of the fucosyltransferase 2 (FUT2), FUT3, TLR5, TAP2, IL1β genes in the duodenum, indicating that these genes probably play a crucial role in the resistance to E. coli F18
Our previous studies concentrated on the relationship of immune gene expression and E. coli F18 resistance in pigs [4,5,6,7,8], only a few reports directly on receptor formation showing the effect of the α(1,2) fucosyltransferase 1 (FUT1) gene M307 G/A mutation on the adhesion of E. coli F18 to pig intestinal epithelial cells [9], but its polymorphism distribution in more than 20 Chinese local pig breeds and wild boar population is extremely skewed [10,11,12]
Summary
Porcine post-weaning diarrhea (PWD) causes serious economic losses to large-scale pig farms. Our previous studies concentrated on the relationship of immune gene expression and E. coli F18 resistance in pigs [4,5,6,7,8], only a few reports directly on receptor formation showing the effect of the α(1,2) fucosyltransferase 1 (FUT1) gene M307 G/A mutation on the adhesion of E. coli F18 to pig intestinal epithelial cells [9], but its polymorphism distribution in more than 20 Chinese local pig breeds and wild boar population is extremely skewed [10,11,12]. We used bisulfite amplicon sequencing (BSAS) to determine the methylation levels of CpG islands in the FUT2 promoter region in intestinal tissues of weaned piglets. This study revealed the regulatory mechanism of FUT2 expression controlling E. coli F18 resistance in weaned piglets, and provided the basis of strategies for the bioengineering regulation of E. coli F18 resistance in human and pigs
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