Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells
Japanese encephalitis virus (JEV) remains the predominant cause of viral encephalitis worldwide. It reaches the central nervous system upon crossing the blood–brain barrier through pathogenic mechanisms that are not completely understood. Here, using a high-throughput siRNA screening assay combined...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-01-01
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| Series: | Emerging Microbes and Infections |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2020.1757388 |
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| author | Yan-Gang Liu Yang Chen Xiaohang Wang Ping Zhao Yongzhe Zhu Zhongtian Qi |
| author_facet | Yan-Gang Liu Yang Chen Xiaohang Wang Ping Zhao Yongzhe Zhu Zhongtian Qi |
| author_sort | Yan-Gang Liu |
| collection | DOAJ |
| description | Japanese encephalitis virus (JEV) remains the predominant cause of viral encephalitis worldwide. It reaches the central nervous system upon crossing the blood–brain barrier through pathogenic mechanisms that are not completely understood. Here, using a high-throughput siRNA screening assay combined with verification experiments, we found that JEV enters the primary human brain microvascular endothelial cells (HBMEC) through a caveolae-mediated endocytic pathway. The role of ezrin, an essential host factor for JEV entry based on our screening, in caveolae-mediated JEV internalization was investigated. We observed that JEV internalization in HBMEC is largely dependent on ezrin-mediated actin cytoskeleton polymerization. Moreover, Src, a protein predicted by a STRING database search, was found to be required in JEV entry. By a variety of pharmacological inhibition and immunoprecipitation assays, we found that Src, ezrin, and caveolin-1 were sequentially activated and formed a complex during JEV infection. A combination of in vitro kinase assay and subcellular analysis demonstrated that ezrin is essential for Src-caveolin-1 interactions. In vivo, both Src and ezrin inhibitors protected ICR suckling mice against JEV-induced mortality and diminished mouse brain viral load. Therefore, JEV entry into HBMEC requires the activation of the Src-ezrin-caveolin-1 signalling axis, which provides potential targets for restricting JEV infection. |
| format | Article |
| id | doaj-art-ee3d7730ed2c4331a89d183fc5a41822 |
| institution | OA Journals |
| issn | 2222-1751 |
| language | English |
| publishDate | 2020-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj-art-ee3d7730ed2c4331a89d183fc5a418222025-08-20T02:26:30ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512020-01-01911330134110.1080/22221751.2020.1757388Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cellsYan-Gang Liu0Yang Chen1Xiaohang Wang2Ping Zhao3Yongzhe Zhu4Zhongtian Qi5Department of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaDepartment of Microbiology, Shanghai Key Laboratory of Medical Biodefense, Naval Medical University (Second Military Medical University), Shanghai, People’s Republic of ChinaJapanese encephalitis virus (JEV) remains the predominant cause of viral encephalitis worldwide. It reaches the central nervous system upon crossing the blood–brain barrier through pathogenic mechanisms that are not completely understood. Here, using a high-throughput siRNA screening assay combined with verification experiments, we found that JEV enters the primary human brain microvascular endothelial cells (HBMEC) through a caveolae-mediated endocytic pathway. The role of ezrin, an essential host factor for JEV entry based on our screening, in caveolae-mediated JEV internalization was investigated. We observed that JEV internalization in HBMEC is largely dependent on ezrin-mediated actin cytoskeleton polymerization. Moreover, Src, a protein predicted by a STRING database search, was found to be required in JEV entry. By a variety of pharmacological inhibition and immunoprecipitation assays, we found that Src, ezrin, and caveolin-1 were sequentially activated and formed a complex during JEV infection. A combination of in vitro kinase assay and subcellular analysis demonstrated that ezrin is essential for Src-caveolin-1 interactions. In vivo, both Src and ezrin inhibitors protected ICR suckling mice against JEV-induced mortality and diminished mouse brain viral load. Therefore, JEV entry into HBMEC requires the activation of the Src-ezrin-caveolin-1 signalling axis, which provides potential targets for restricting JEV infection.https://www.tandfonline.com/doi/10.1080/22221751.2020.1757388Brain microvascular endothelial cellscaveolin-1ezrinJapanese encephalitis virusSrcviral entry |
| spellingShingle | Yan-Gang Liu Yang Chen Xiaohang Wang Ping Zhao Yongzhe Zhu Zhongtian Qi Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells Emerging Microbes and Infections Brain microvascular endothelial cells caveolin-1 ezrin Japanese encephalitis virus Src viral entry |
| title | Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells |
| title_full | Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells |
| title_fullStr | Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells |
| title_full_unstemmed | Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells |
| title_short | Ezrin is essential for the entry of Japanese encephalitis virus into the human brain microvascular endothelial cells |
| title_sort | ezrin is essential for the entry of japanese encephalitis virus into the human brain microvascular endothelial cells |
| topic | Brain microvascular endothelial cells caveolin-1 ezrin Japanese encephalitis virus Src viral entry |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2020.1757388 |
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