Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays
Throughout their life cycle, viruses interact with cellular host factors, thereby influencing propagation, host range, cell tropism and pathogenesis. The hepatitis E virus (HEV) is an underestimated RNA virus in which knowledge of the virus–host interaction network to date is limited. Here, two rela...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-12-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/15/12/2412 |
| _version_ | 1797379068181610496 |
|---|---|
| author | Laura Corneillie Irma Lemmens Karin Weening Amse De Meyer Freya Van Houtte Jan Tavernier Philip Meuleman |
| author_facet | Laura Corneillie Irma Lemmens Karin Weening Amse De Meyer Freya Van Houtte Jan Tavernier Philip Meuleman |
| author_sort | Laura Corneillie |
| collection | DOAJ |
| description | Throughout their life cycle, viruses interact with cellular host factors, thereby influencing propagation, host range, cell tropism and pathogenesis. The hepatitis E virus (HEV) is an underestimated RNA virus in which knowledge of the virus–host interaction network to date is limited. Here, two related high-throughput mammalian two-hybrid approaches (MAPPIT and KISS) were used to screen for HEV-interacting host proteins. Promising hits were examined on protein function, involved pathway(s), and their relation to other viruses. We identified 37 ORF2 hits, 187 for ORF3 and 91 for ORF4. Several hits had functions in the life cycle of distinct viruses. We focused on SHARPIN and RNF5 as candidate hits for ORF3, as they are involved in the RLR-MAVS pathway and interferon (IFN) induction during viral infections. Knocking out (KO) SHARPIN and RNF5 resulted in a different IFN response upon ORF3 transfection, compared to wild-type cells. Moreover, infection was increased in SHARPIN KO cells and decreased in RNF5 KO cells. In conclusion, MAPPIT and KISS are valuable tools to study virus–host interactions, providing insights into the poorly understood HEV life cycle. We further provide evidence for two identified hits as new host factors in the HEV life cycle. |
| first_indexed | 2024-03-08T20:16:49Z |
| format | Article |
| id | doaj.art-23e11388b4bb41cb8d0c3e7a62c97abd |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-08T20:16:49Z |
| publishDate | 2023-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-23e11388b4bb41cb8d0c3e7a62c97abd2023-12-22T14:49:22ZengMDPI AGViruses1999-49152023-12-011512241210.3390/v15122412Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid AssaysLaura Corneillie0Irma Lemmens1Karin Weening2Amse De Meyer3Freya Van Houtte4Jan Tavernier5Philip Meuleman6Laboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumVIB-UGent Center for Medical Biotechnology, Department of Biomolecular Medicine, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumLaboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumLaboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumLaboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumVIB-UGent Center for Medical Biotechnology, Department of Biomolecular Medicine, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumLaboratory of Liver Infectious Diseases, Department of Diagnostic Sciences, Faculty of Medicine and Health Sciences, Ghent University, 9000 Ghent, BelgiumThroughout their life cycle, viruses interact with cellular host factors, thereby influencing propagation, host range, cell tropism and pathogenesis. The hepatitis E virus (HEV) is an underestimated RNA virus in which knowledge of the virus–host interaction network to date is limited. Here, two related high-throughput mammalian two-hybrid approaches (MAPPIT and KISS) were used to screen for HEV-interacting host proteins. Promising hits were examined on protein function, involved pathway(s), and their relation to other viruses. We identified 37 ORF2 hits, 187 for ORF3 and 91 for ORF4. Several hits had functions in the life cycle of distinct viruses. We focused on SHARPIN and RNF5 as candidate hits for ORF3, as they are involved in the RLR-MAVS pathway and interferon (IFN) induction during viral infections. Knocking out (KO) SHARPIN and RNF5 resulted in a different IFN response upon ORF3 transfection, compared to wild-type cells. Moreover, infection was increased in SHARPIN KO cells and decreased in RNF5 KO cells. In conclusion, MAPPIT and KISS are valuable tools to study virus–host interactions, providing insights into the poorly understood HEV life cycle. We further provide evidence for two identified hits as new host factors in the HEV life cycle.https://www.mdpi.com/1999-4915/15/12/2412protein–protein interactionsMAPPITKISSviral hepatitishost factorhepatitis E virus |
| spellingShingle | Laura Corneillie Irma Lemmens Karin Weening Amse De Meyer Freya Van Houtte Jan Tavernier Philip Meuleman Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays Viruses protein–protein interactions MAPPIT KISS viral hepatitis host factor hepatitis E virus |
| title | Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays |
| title_full | Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays |
| title_fullStr | Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays |
| title_full_unstemmed | Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays |
| title_short | Virus–Host Protein Interaction Network of the Hepatitis E Virus ORF2-4 by Mammalian Two-Hybrid Assays |
| title_sort | virus host protein interaction network of the hepatitis e virus orf2 4 by mammalian two hybrid assays |
| topic | protein–protein interactions MAPPIT KISS viral hepatitis host factor hepatitis E virus |
| url | https://www.mdpi.com/1999-4915/15/12/2412 |
| work_keys_str_mv | AT lauracorneillie virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT irmalemmens virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT karinweening virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT amsedemeyer virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT freyavanhoutte virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT jantavernier virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays AT philipmeuleman virushostproteininteractionnetworkofthehepatitisevirusorf24bymammaliantwohybridassays |