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The mechanism of HEV replication, especially host factors governing HEV replication is still not clear. Here, using HEV ORF1 <jats:italic>trans<\/jats:italic>-complementation cell culture system and HEV replicon system, combining with stable isotope labelling with amino acids in cell culture (SILAC) and mass spectrometry (MS), we aimed to identify the host factors regulating HEV replication. We identified a diversity of host factors associated with HEV ORF1 protein, which were putatively responsible for viral genomic RNA replication, in these two cell culture models. Of note, the protein arginine methyltransferase 5 (PRMT5)\/WDR77 complex was identified in both cell culture models as the top hit. Furthermore, we demonstrated that PRMT5 and WDR77 can specifically inhibit HEV replication, but not other viruses such as HCV or SARS-CoV-2, and this inhibition is conserved among different HEV strains and genotypes. Mechanistically, PRMT5\/WDR77 can catalyse methylation of ORF1 on its R458, impairing its replicase activity, and virus bearing R458K mutation in ORF1 relieves the restriction of PRMT5\/WDR77 accordingly. Taken together, our study promotes more comprehensive understanding of viral infections but also provides therapeutic targets for intervention.<\/jats:p>","DOI":"10.1371\/journal.ppat.1011434","type":"journal-article","created":{"date-parts":[[2023,6,5]],"date-time":"2023-06-05T17:26:06Z","timestamp":1685985966000},"page":"e1011434","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":11,"title":["The PRMT5\/WDR77 complex restricts hepatitis E virus replication"],"prefix":"10.1371","volume":"19","author":[{"given":"Xiaohui","family":"Ju","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanying","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenlin","family":"Ren","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Dong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianbin","family":"Meng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haiteng","family":"Deng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuchen","family":"Nan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6226-869X","authenticated-orcid":true,"given":"Qiang","family":"Ding","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,6,5]]},"reference":[{"issue":"4","key":"ppat.1011434.ref001","doi-asserted-by":"crossref","first-page":"988","DOI":"10.1002\/hep.25505","article-title":"The global burden of hepatitis E virus genotypes 1 and 2 in 2005","volume":"55","author":"DB Rein","year":"2012","journal-title":"Hepatology"},{"issue":"7","key":"ppat.1011434.ref002","doi-asserted-by":"crossref","first-page":"1516","DOI":"10.1111\/liv.14468","article-title":"The global epidemiology of hepatitis E virus infection: A systematic review and meta-analysis","volume":"40","author":"P Li","year":"2020","journal-title":"Liver Int"},{"issue":"1","key":"ppat.1011434.ref003","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.virusres.2011.03.006","article-title":"Hepatitis E virus infection in developed countries","volume":"161","author":"T. 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