title: Biological or pharmacological activation of protein kinase C alpha constrains hepatitis E virus replication.
abstract: Although hepatitis E has emerged as a global health issue, there is limited knowledge of its infection biology and no FDA-approved medication is available. Aiming to investigate the role of protein kinases in hepatitis E virus (HEV) infection and to identify potential antiviral targets, we screened a library of pharmacological kinase inhibitors in a cell culture model, a subgenomic HEV replicon containing luciferase reporter. We identified protein kinase C alpha (PKCα) as an essential cell host factor restricting HEV replication. Both specific inhibitor and shRNA-mediated knockdown of PKCα enhanced HEV replication. Conversely, over-expression of the activated form of PKCα or treatment with its pharmacological activator strongly inhibited HEV replication. Interestingly, upon the stimulation by its activator, PKCα efficiently activates its downstream Activator Protein 1 (AP-1) pathway, leading to the induction of antiviral interferon-stimulated genes (ISGs). This process is independent of the JAK-STAT machinery and interferon production. However, PKCα induced HEV inhibition appears independent of the AP1 cascade. The discovery that activated PKCα restricts HEV replication reveals new insight of HEV-host interactions and provides new target for antiviral drug development.
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Wenshi
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Yijin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Debing
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Xinying
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Yuebang
authors.initials: Y
authors.lastname: Yin
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Lei
authors.initials: L
authors.lastname: Xu
authors.affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Elena
authors.initials: E
authors.lastname: Herrera Carrillo
authors.affiliation: Department of Cell Biology, Medical Genetics Cluster, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Johannes H
authors.initials: JH
authors.lastname: Brandsma
authors.affiliation: Department of Cell Biology, Medical Genetics Cluster, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Raymond A
authors.initials: RA
authors.lastname: Poot
authors.affiliation: Laboratory of Experimental Virology, Department of Medical Microbiology, Center for Infection and Immunity Amsterdam (CINIMA), Academic Medical Center of the University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Ben
authors.initials: B
authors.lastname: Berkhout
authors.affiliation: Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven, Leuven, Belgium.
authors.forename: Johan
authors.initials: J
authors.lastname: Neyts
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
authors.forename: Maikel P
authors.initials: MP
authors.lastname: Peppelenbosch
authors.affiliation: Department of Gastroenterology and Hepatology, Postgraduate School Molecular Medicine, Erasmus MC, University Medical Center, Rotterdam, The Netherlands. Electronic address: q.pan@erasmusmc.nl.
authors.forename: Qiuwei
authors.initials: Q
authors.lastname: Pan
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification / Q000494:pharmacology*; D018929:Cell Culture Techniques; D004261:DNA Replication; D004789:Enzyme Activation; D016751:Hepatitis E / Q000188:drug therapy; D016752:Hepatitis E virus / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D051571:Protein Kinase C-alpha / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D047428:Protein Kinase Inhibitors / Q000302:isolation & purification / Q000494:pharmacology*; D012093:Replicon; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018808:Transcription Factor AP-1 / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000998:Antiviral Agents; D047428:Protein Kinase Inhibitors; D018808:Transcription Factor AP-1; C496628:PRKCA protein, human; D051571:Protein Kinase C-alpha; D013755:Tetradecanoylphorbol Acetate
keywords: AP-1; Hepatitis E virus; ISG; PKCα; PMA