The m6A demethylase ALKBH5 resists PEDV infection by regulating GAS6 expression in porcine alveolar macrophages

Porcine epidemic diarrhea virus (PEDV) causes porcine epidemic diarrhea (PED), which is a highly contagious intestinal infectious disease characterized by vomiting, diarrhea and dehydration and high mortality in suckling piglets. N6-methyladenosine (m6A) is the most common mode of modification in eukaryotic mRNAs, and m6A modification has an important regulatory role in the process of viral replication and host immunity. However, the role and molecular mechanism of m6A modification on PEDV infection resistance in pigs are not fully understood. In this study, we first examined the effect of PEDV infection on different m6A modifying enzymes at the tissue and cellular levels, determined the key m6A modifying enzyme ALKBH5, and further verified that down-regulation of ALKBH5 expression levels was not conducive to 3D4/21 cells to resist PEDV infection using RNA interference. We determined the genome-wide gene expression alterations between sh-ALKBH5 and sh-NC groups using RNA-Seq, which identified a total of 175 differentially expressed genes (DEGs), of which 97 were down-regulated and 78 were up-regulated. Based on the results of RNA-Seq, we initially regard GAS6 as a candidate gene for possible targeted regulation of ALKBH5.