The endocytic recycling compartment serves as a viral factory for hepatitis E virus.
AlphaFold2
Antibodies
Electron microscopy
Endocytic recycling compartment
Hepatitis E virus
Infectious particles
ORF2 capsid protein
Rab11
Viral factories
Journal
Cellular and molecular life sciences : CMLS
ISSN: 1420-9071
Titre abrégé: Cell Mol Life Sci
Pays: Switzerland
ID NLM: 9705402
Informations de publication
Date de publication:
03 Dec 2022
03 Dec 2022
Historique:
received:
26
08
2022
accepted:
23
11
2022
revised:
04
11
2022
entrez:
2
12
2022
pubmed:
3
12
2022
medline:
7
12
2022
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 36460928
doi: 10.1007/s00018-022-04646-y
pii: 10.1007/s00018-022-04646-y
pmc: PMC9718719
doi:
Substances chimiques
Capsid Proteins
0
Antibodies, Monoclonal
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
615Subventions
Organisme : Inserm Transfert
ID : MAT-PI-17006
Organisme : Institut Pasteur de Lille
ID : DiagHepE
Organisme : Région Hauts-de-France
ID : DiagHepE
Informations de copyright
© 2022. The Author(s).
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