The endocytic recycling compartment serves as a viral factory for hepatitis E virus

Author:

Bentaleb Cyrine,Hervouet Kévin,Montpellier Claire,Camuzet Charline,Ferrié Martin,Burlaud-Gaillard Julien,Bressanelli Stéphane,Metzger Karoline,Werkmeister Elisabeth,Ankavay Maliki,Janampa Nancy Leon,Marlet Julien,Roux Julien,Deffaud Clarence,Goffard Anne,Rouillé Yves,Dubuisson Jean,Roingeard Philippe,Aliouat-Denis Cécile-Marie,Cocquerel LaurenceORCID

Abstract

AbstractAlthough hepatitis E virus (HEV) is the major leading cause of enterically transmitted viral hepatitis worldwide, many gaps remain in the understanding of the HEV lifecycle. Notably, viral factories induced by HEV have not been documented yet, and it is currently unknown whether HEV infection leads to cellular membrane modeling as many positive-strand RNA viruses. HEV genome encodes the ORF1 replicase, the ORF2 capsid protein and the ORF3 protein involved in virion egress. Previously, we demonstrated that HEV produces different ORF2 isoforms including the virion-associated ORF2i form. Here, we generated monoclonal antibodies that specifically recognize the ORF2i form and antibodies that recognize the different ORF2 isoforms. One antibody, named P1H1 and targeting the ORF2i N-terminus, recognized delipidated HEV particles from cell culture and patient sera. Importantly, AlphaFold2 modeling demonstrated that the P1H1 epitope is exposed on HEV particles. Next, antibodies were used to probe viral factories in HEV-producing/infected cells. By confocal microscopy, we identified subcellular nugget-like structures enriched in ORF1, ORF2 and ORF3 proteins and viral RNA. Electron microscopy analyses revealed an unprecedented HEV-induced membrane network containing tubular and vesicular structures. We showed that these structures are dependent on ORF2i capsid protein assembly and ORF3 expression. An extensive colocalization study of viral proteins with subcellular markers, and silencing experiments demonstrated that these structures are derived from the endocytic recycling compartment (ERC) for which Rab11 is a central player. Hence, HEV hijacks the ERC and forms a membrane network of vesicular and tubular structures that might be the hallmark of HEV infection.

Funder

Inserm Transfert

Institut Pasteur de Lille

Région Hauts-de-France

Agence Nationale de Recherches sur le Sida et les Hépatites Virales

Université de Tours

Institut Pasteur De Lille

Publisher

Springer Science and Business Media LLC

Subject

Cell Biology,Cellular and Molecular Neuroscience,Pharmacology,Molecular Biology,Molecular Medicine

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