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Cited by 27Hepatitis Viruses Studies and EpidemiologyCellular transport and secretionHepatitis B Virus Studies

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

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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, Laurence·2022·Cellular and Molecular Life Sciences
DOI 10.1007/s00018-022-04646-yPMC PMC9718719

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Abstract

Although 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.

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Metadata

Authors
Bentaleb, C., Hervouet, K., Montpellier, C., Camuzet, C., Ferrié, M., Burlaud-Gaillard, J., Bressanelli, S., Metzger, K., Werkmeister, E., Ankavay, M., Janampa, N. L., Marlet, J., Roux, J., Deffaud, C., Goffard, A., Rouillé, Y., Dubuisson, J., Roingeard, P., Aliouat-Denis, C.-M., & Cocquerel, L.
Affiliations
  • Centre National de la Recherche Scientifique (FR)
  • Université de Tours (FR)
Corresponding author
Cocquerel, Laurence
ORCID
Venue
Cellular and Molecular Life Sciences
Publisher
Springer Science and Business Media LLC
Published
2022-12
Volume
79
Issue
12
Pages
615-615
ISSN
  • 1420-682X (ISSN-L)
  • 1420-9071
DOI
10.1007/s00018-022-04646-y
PMC
PMC9718719
OpenAlex
W4311179475
Cited by
27
References
1
Citations by year
2026: 7 · 2025: 8 · 2024: 3 · 2023: 4 · 2022: 5
FWCI
2.98
Type
journal article
Language
en
Open access
Licence
Locations
  • Cellular and Molecular Life Sciences (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • SERVAL (Université de Lausanne) (publishedVersion, cc-by)
  • HAL (submittedVersion)
  • HAL (submittedVersion, other-oa)
  • IRIS (submittedVersion, cc-by)
  • LillOA (Université de Lille (University Of Lille)) (submittedVersion, other-oa)
Funders
  • Inserm Transfert (MAT-PI-17006)
  • Institut Pasteur de Lille (DiagHepE)
  • Région Hauts-de-France (DiagHepE)
  • Agence Nationale de Recherches sur le Sida et les Hépatites Virales
  • Université de Tours
Topics
  • Health Sciences › Medicine › Hepatology › Hepatitis Viruses Studies and Epidemiology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Health Sciences › Medicine › Epidemiology › Hepatitis B Virus Studies
Keywords
hepatitis E virus, viral factories, recycling endosomes, Rab11, ORF2 capsid protein, ORF3 protein, monoclonal antibodies, epitope mapping, viral assembly, positive-strand RNA viruses, viral life cycle
MeSH terms
Viral Replication Compartments, Antibodies, Monoclonal, Biological Transport, Humans, Hepatitis E virus, Capsid Proteins

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