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Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle

Ebola virus (EBOV) causes severe EBOV disease (EVD) in humans and non-human primates. Currently, limited countermeasures are available, and the virus must be studied in biosafety level-4 (BSL-4) laboratories. EBOV glycoprotein (GP) is a single transmembrane protein responsible for entry into host ce...

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Autores principales: Furuyama, Wakako, Sakaguchi, Miako, Yamada, Kento, Nanbo, Asuka
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9669576/
https://www.ncbi.nlm.nih.gov/pubmed/36406413
http://dx.doi.org/10.3389/fmicb.2022.1026644
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author Furuyama, Wakako
Sakaguchi, Miako
Yamada, Kento
Nanbo, Asuka
author_facet Furuyama, Wakako
Sakaguchi, Miako
Yamada, Kento
Nanbo, Asuka
author_sort Furuyama, Wakako
collection PubMed
description Ebola virus (EBOV) causes severe EBOV disease (EVD) in humans and non-human primates. Currently, limited countermeasures are available, and the virus must be studied in biosafety level-4 (BSL-4) laboratories. EBOV glycoprotein (GP) is a single transmembrane protein responsible for entry into host cells and is the target of multiple approved drugs. However, the molecular mechanisms underlying the intracellular dynamics of GP during EBOV lifecycle are poorly understood. In this study, we developed a novel GP monitoring system using transcription- and replication-competent virus-like particles (trVLPs) that enables the modeling of the EBOV lifecycle under BSL-2 conditions. We constructed plasmids to generate trVLPs containing the coding sequence of EBOV GP, in which the mucin-like domain (MLD) was replaced with fluorescent proteins. The generated trVLP efficiently replicated over multiple generations was similar to the wild type trVLP. Furthermore, we confirmed that the novel trVLP system enabled real-time visualization of GP throughout the trVLP replication cycle and exhibited intracellular localization similar to that of wild type GP. In summary, this novel monitoring system for GP will enable the characterization of the molecular mechanism of the EBOV lifecycle and can be applied for the development of therapeutics against EVD.
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spelling pubmed-96695762022-11-18 Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle Furuyama, Wakako Sakaguchi, Miako Yamada, Kento Nanbo, Asuka Front Microbiol Microbiology Ebola virus (EBOV) causes severe EBOV disease (EVD) in humans and non-human primates. Currently, limited countermeasures are available, and the virus must be studied in biosafety level-4 (BSL-4) laboratories. EBOV glycoprotein (GP) is a single transmembrane protein responsible for entry into host cells and is the target of multiple approved drugs. However, the molecular mechanisms underlying the intracellular dynamics of GP during EBOV lifecycle are poorly understood. In this study, we developed a novel GP monitoring system using transcription- and replication-competent virus-like particles (trVLPs) that enables the modeling of the EBOV lifecycle under BSL-2 conditions. We constructed plasmids to generate trVLPs containing the coding sequence of EBOV GP, in which the mucin-like domain (MLD) was replaced with fluorescent proteins. The generated trVLP efficiently replicated over multiple generations was similar to the wild type trVLP. Furthermore, we confirmed that the novel trVLP system enabled real-time visualization of GP throughout the trVLP replication cycle and exhibited intracellular localization similar to that of wild type GP. In summary, this novel monitoring system for GP will enable the characterization of the molecular mechanism of the EBOV lifecycle and can be applied for the development of therapeutics against EVD. Frontiers Media S.A. 2022-11-03 /pmc/articles/PMC9669576/ /pubmed/36406413 http://dx.doi.org/10.3389/fmicb.2022.1026644 Text en Copyright © 2022 Furuyama, Sakaguchi, Yamada and Nanbo. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Furuyama, Wakako
Sakaguchi, Miako
Yamada, Kento
Nanbo, Asuka
Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title_full Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title_fullStr Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title_full_unstemmed Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title_short Development of an imaging system for visualization of Ebola virus glycoprotein throughout the viral lifecycle
title_sort development of an imaging system for visualization of ebola virus glycoprotein throughout the viral lifecycle
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9669576/
https://www.ncbi.nlm.nih.gov/pubmed/36406413
http://dx.doi.org/10.3389/fmicb.2022.1026644
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