DNA Methylation of Pig FUT3 Promoter Alters mRNA Expression to Regulate E. coli F18 Susceptibility

Abstract

Post-weaning diarrhea (PWD) is frequently associated with E. coli F18 infections in piglets. However, the underlying molecular mechanism concerning the resistance of E. coli F18 in local weaned piglets in China is not clearly understood. In the present study, by a comparative analysis of the transcriptome, a-1,3-fucosyltransferase (FUT3) was evaluated as a key candidate correlated with resistance to E. coli F18 in Sutai and Meishan piglets. Functional analysis demonstrated that FUT3 acts as a key positive regulator of E. coli F18 susceptibility in newly food accustomed piglets. However, the core promoter of FUT3 was present at −500–(−206) bp (chr.2: g.73171117–g.73171616), comprising of 9 methylated CpG sites. Among these, the methylation levels of the two CpG sites (mC-3, mC-5) located in HIF1A and Sp1 transcription factor (TF) considerably associated with mRNA expression of FUT3 (p < 0.05). Our findings indicated that the methylation of mC-3 and mC-5 sites has certain inhibitory effect on FUT3 expression and promotes the resistance of E. coli F18 in piglets. The underlined study may explore FUT3 as a new candidate target in E. coli F18 infection in Chinese local weaned piglets.

Insight into mechanisms of pig lncRNA FUT3-AS1 regulating E. coli F18-bacterial diarrhea

“…As shown in Fig 3G , histone H4 levels were increased significantly in the promoter regions of FUT3 compared with the controls ( P <0.01). Besides, we have identified the core promoter region (chr2: g.73171117–g.73171616) of FUT3 in previous study [ 32 ]. On this basis, we performed a DNA-pull down assay and identified 173 specific FUT3 promoter-associated proteins using mass spectrometry, including histone H4.…”

“…Additionally, to explore the mechanism of DNA methylation regulating FUT3 gene expression, we also determined the core promoter (chr.2: g.73171117-g.73171616) of FUT3 gene, and found 9 methylated CpG sites in the core promoter by bisulfifite amplicon sequencing (BSAS). Further analysis showed the methylation levels of two CpG sites (mC-3, mC-5) located in HIF1A and Sp1 transcription factor have certain inhibitory effect on FUT3 expression [ 32 ]. In this study, our findings revealed that the lncRNA regulated FUT3 expression via H4K16ac modification or ceRNA mechanism, and systematic experiments demonstrated that FUT3 acts as a positive regulator of E .…”

Comprehensive Analysis Revealed the Potential Roles of N6-Methyladenosine (m6A) Mediating E. coli F18 Susceptibility in IPEC-J2 Cells

“…Thus, resistance against the E. coli F18 invasion depends on the intestinal immunity and F18-receptor content in pigs. Recently, several studies analyzed the association between immune-related genes ( CD14, TLR5 ) [ 4 , 5 ], or fucosetransferase genes ( FUT2, FUT3 ) [ 6 , 7 ] and E. coli F18 susceptibility in piglets. Nevertheless, the mechanism behind piglets’ resistance to E. coli F18 invasion is not fully understood.…”

“…In general, the intestinal porcine epithelial cell (IPEC-J2) is an efficient cell model for the examination of the underlying mechanism behind the host anti- E. coli F18 pathogen [ 7 , 20 ]. In addition, multiple studies have revealed that the IPEC-J2 exposure to E. coli triggers oxidative damage and inflammatory responses, which results in enhancing apoptotic cell death and diminishing functionality [ 21 , 22 , 23 ].…”