CROWN-seq profiling of snRNA m6Am modification in pig ST cells upon PRV infection

Fat mass and obesity-associated protein (FTO) was discovered as the first RNA demethylase and mediates the demethylation of N6,2'-O-dimethyladenosine (m6Am), a prevalent RNA modification adjacent to the 5' cap. Despite its significance, the regulation of the m6Am demethylase and the potential impact of virus infection on FTO remain largely unexplored. In this study, we show that the alphaherpesvirus pseudorabies virus (PRV) triggers phosphorylation of FTO and that this phosphorylation is caused by the viral serine/threonine protein kinase UL13. To test whether the phosphorylated FTO can result in different demethylation on snRNA, we performed CROWN-seq to quantify m6Am stoichiometry in cells. Overall design: Three replicates were performed on ST cells (mock), ST cells transfected by wild-type PRV, and ST cells transfected by UL13-null PRV