title: Hepatitis E virus inhibits type I interferon induction by ORF1 products.
abstract: Hepatitis E virus (HEV) causes both endemic and epidemic human hepatitis by fecal-oral transmission in many parts of the world. Zoonotic transmission of HEV from animals to humans has been reported. Due to the lack of an efficient cell culture system, the molecular mechanisms of HEV infection remain largely unknown. In this study, we found that HEV replication in hepatoma cells inhibited poly(I·C)-induced beta interferon (IFN-β) expression and that the HEV open reading frame 1 (ORF1) product was responsible for this inhibition. Two domains, X and the papain-like cysteine protease domain (PCP), of HEV ORF1 were identified as the putative IFN antagonists. When overexpressed in HEK293T cells, the X domain (or macro domain) inhibited poly(I·C)-induced phosphorylation of interferon regulatory factor 3 (IRF-3), which is the key transcription factor for IFN induction. The PCP domain was shown to have deubiquitinase activity for both RIG-I and TBK-1, whose ubiquitination is a key step in their activation in poly(I·C)-induced IFN induction. Furthermore, replication of a HEV replicon containing green fluorescent protein (GFP) (E2-GFP) in hepatoma cells led to impaired phosphorylation of IRF-3 and reduced ubiquitination of RIG-I and TBK-1, which confirmed our observations of X and PCP inhibitory effects in HEK293T cells. Altogether, our study identified the IFN antagonists within the HEV ORF1 polyprotein and expanded our understanding of the functions of several of the HEV ORF1 products, as well as the mechanisms of HEV pathogenesis. Importance: Type I interferons (IFNs) are important components of innate immunity and play a crucial role against viral infection. They also serve as key regulators to evoke an adaptive immune response. Virus infection can induce the synthesis of interferons; however, viruses have evolved many strategies to antagonize the induction of interferons. There is little knowledge about how hepatitis E virus (HEV) inhibits induction of host IFNs, though the viral genome was sequenced more than 2 decades ago. This is the first report of identification of the potential IFN antagonists encoded by HEV. By screening all the domains in the open reading frame 1 (ORF1) polyprotein, we identified two IFN antagonists and performed further research to determine how and at which step in the IFN induction pathway they antagonize host IFN induction. Our work provides valuable information about HEV-cell interaction and pathogenesis.
authors.affiliation: Molecular Virology Laboratory, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Nan
authors.affiliation: Molecular Virology Laboratory, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Yu
authors.affiliation: Molecular Virology Laboratory, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA.
authors.forename: Zexu
authors.initials: Z
authors.lastname: Ma
authors.affiliation: Virology Laboratory, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Khattar
authors.affiliation: Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, Maryland, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Fredericksen
authors.affiliation: Molecular Virology Laboratory, Virginia-Maryland College of Veterinary Medicine and Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA zhangyj@umd.edu.
authors.forename: Yan-Jin
authors.initials: YJ
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D057809:HEK293 Cells; D016752:Hepatitis E virus / Q000502:physiology*; D006801:Humans; D007370:Interferon Type I / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D016366:Open Reading Frames; D010766:Phosphorylation; D054875:Ubiquitination; D014764:Viral Proteins / Q000502:physiology
chemical_list: D017931:DNA Primers; D007370:Interferon Type I; D014764:Viral Proteins
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