Hepatitis E virus (HEV) infects ~20 million people annually, causing 70,000 deaths. HEV has a positive sense RNA genome that contains 3 conserved ORFs, of these ORF1 encodes a large non-structural polyprotein (pORF1) necessary and essential for viral genome replication.  The study of pORF1 functions and virus genome replication has been hampered by a paucity of reagents. To address this we previously used transposon mutagenesis to identify two insertion sites within pORF1 that tolerated epitope tagging without compromising the replication of an HEV subgenomic replicon (SGR). Subsequently, we used these novel SGRs expressing tagged pORF1 proteins to establish a stable model of viral persistence. Immunoprecipitation of pORF1 coupled with tandem mass spectrometry identified 146 host proteins as putative novel targets for pORF1. Notably, one of these was fatty acid synthase (FASN), and given the important role of lipid metabolism in positive-sense RNA virus replication, we investigated the role of FASN in HEV replication.  HEV genome replication was inhibited by pharmacological inhibition of both FASN and protein palmitoylation, suggesting that FASN-derived palmitate supports pORF1 post-translational modification and viral replication. Ongoing experiments aim to determine how FASN influences HEV pORF1 synthesis, whether FASN-derived palmitate is required for replication complex formation, and viral protein palmitoylation. Understanding this mechanism will reveal HEV-host metabolic dependencies and guide host-targeted antiviral development.