Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles. Author Qiang Ding, Brigitte Heller, Juan Capuccino, Bokai Song, Ila Nimgaonkar, Gabriela Hrebikova, Jorge Contreras, Alexander Ploss Publication Year 2017 Type Journal Article Abstract Hepatitis E virus (HEV) is the leading cause of enterically transmitted viral hepatitis globally. Of HEV's three ORFs, the function of ORF3 has remained elusive. Here, we demonstrate that via homophilic interactions ORF3 forms multimeric complexes associated with intracellular endoplasmic reticulum (ER)-derived membranes. HEV ORF3 shares several structural features with class I viroporins, and the function of HEV ORF3 can be maintained by replacing it with the well-characterized viroporin influenza A virus (IAV) matrix-2 protein. ORF3's ion channel function is further evidenced by its ability to mediate ionic currents when expressed in Xenopus laevis oocytes. Furthermore, we identified several positions in ORF3 critical for its formation of multimeric complexes, ion channel activity, and, ultimately, release of infectious particles. Collectively, our data demonstrate a previously undescribed function of HEV ORF3 as a viroporin, which may serve as an attractive target in developing direct-acting antivirals. Keywords Animals, Structure-Activity Relationship, Humans, Gene Deletion, Amino Acid Substitution, Recombinant Fusion Proteins, Amino Acid Sequence, Patch-Clamp Techniques, HEK293 Cells, Xenopus laevis, Virus Replication, Viral Proteins, Oocytes, Endoplasmic Reticulum, Amino Acid Motifs, Protein Domains, Ion Channels, Hep G2 Cells, Hepatitis E virus, Ion Transport, Viral Matrix Proteins, Virus Release Journal Proc Natl Acad Sci U S A Volume 114 Issue 5 Pages 1147-1152 Date Published 2017 Jan 31 ISSN Number 1091-6490 DOI 10.1073/pnas.1614955114 Alternate Journal Proc Natl Acad Sci U S A PMCID PMC5293053 PMID 28096411 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML