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Significance Hepatitis E virus (HEV) is responsible for an estimated 20 million enterically transmitted cases of viral hepatitis globally. Here, we demonstrate that one of HEV’s three major gene products, ORF3, is an ion channel. Deletion of ORF3 abrogates release of infectious virions, and we show that viral egress can be rescued by expressing the influenza A virus (IAV) matrix-2 proteinin trans . Expression of ORF3 facilitates ion flux across the plasma membrane, providing direct evidence for its viroporin activity. We identify regions within ORF3 abrogating both ion channel activity and particle release, thereby linking these two processes for a quasienveloped human virus and providing an attractive potential target for antiviral drug development.

Hepatitis E virus ORF3 is a functional ion channel required for release of infectious particles