Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization

Viral RNA synthesis of several non-segmented, negative-sense RNA viruses (NNSVs) takes place in inclusion bodies (IBs) that show properties of liquid organelles, which are formed by liquid–liquid phase separation of scaffold proteins. It is believed that this is driven by intrinsically disordered re...

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Main Authors: Bianca S. Bodmer, Melina Vallbracht, Dmitry S. Ushakov, Lisa Wendt, Petr Chlanda, Thomas Hoenen
Format: Article
Language:English
Published: Taylor & Francis Group 2023-12-01
Series:Emerging Microbes and Infections
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Online Access:https://www.tandfonline.com/doi/10.1080/22221751.2023.2223727
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author Bianca S. Bodmer
Melina Vallbracht
Dmitry S. Ushakov
Lisa Wendt
Petr Chlanda
Thomas Hoenen
author_facet Bianca S. Bodmer
Melina Vallbracht
Dmitry S. Ushakov
Lisa Wendt
Petr Chlanda
Thomas Hoenen
author_sort Bianca S. Bodmer
collection DOAJ
description Viral RNA synthesis of several non-segmented, negative-sense RNA viruses (NNSVs) takes place in inclusion bodies (IBs) that show properties of liquid organelles, which are formed by liquid–liquid phase separation of scaffold proteins. It is believed that this is driven by intrinsically disordered regions (IDRs) and/or multiple copies of interaction domains, which for NNSVs are usually located in their nucleo – and phosphoproteins. In contrast to other NNSVs, the Ebola virus (EBOV) nucleoprotein NP alone is sufficient to form IBs without the need for a phosphoprotein, and to facilitate the recruitment of other viral proteins into these structures. While it has been proposed that also EBOV IBs are liquid organelles, this has so far not been formally demonstrated. Here we used a combination of live cell microscopy, fluorescence recovery after photobleaching assays, and mutagenesis approaches together with reverse genetics-based generation of recombinant viruses to study the formation of EBOV IBs. Our results demonstrate that EBOV IBs are indeed liquid organelles, and that oligomerization but not IDRs of the EBOV nucleoprotein plays a key role in their formation. Additionally, VP35 (often considered the phosphoprotein-equivalent of EBOV) is not essential for IB formation, but alters their liquid behaviour. These findings define the molecular mechanism for the formation of EBOV IBs, which play a central role in the life cycle of this deadly virus.
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spelling doaj-art-0710fdb8a91d42d8bb641e184a8ff62c2025-08-20T03:29:35ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512023-12-0112210.1080/22221751.2023.2223727Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerizationBianca S. Bodmer0Melina Vallbracht1Dmitry S. Ushakov2Lisa Wendt3Petr Chlanda4Thomas Hoenen5Institute for Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald - Insel Riems, GermanySchaller Research Groups, Department of Infectious Diseases, Virology, Heidelberg University Hospital, Heidelberg, GermanyInstitute for Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald - Insel Riems, GermanyInstitute for Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald - Insel Riems, GermanySchaller Research Groups, Department of Infectious Diseases, Virology, Heidelberg University Hospital, Heidelberg, GermanyInstitute for Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, Greifswald - Insel Riems, GermanyViral RNA synthesis of several non-segmented, negative-sense RNA viruses (NNSVs) takes place in inclusion bodies (IBs) that show properties of liquid organelles, which are formed by liquid–liquid phase separation of scaffold proteins. It is believed that this is driven by intrinsically disordered regions (IDRs) and/or multiple copies of interaction domains, which for NNSVs are usually located in their nucleo – and phosphoproteins. In contrast to other NNSVs, the Ebola virus (EBOV) nucleoprotein NP alone is sufficient to form IBs without the need for a phosphoprotein, and to facilitate the recruitment of other viral proteins into these structures. While it has been proposed that also EBOV IBs are liquid organelles, this has so far not been formally demonstrated. Here we used a combination of live cell microscopy, fluorescence recovery after photobleaching assays, and mutagenesis approaches together with reverse genetics-based generation of recombinant viruses to study the formation of EBOV IBs. Our results demonstrate that EBOV IBs are indeed liquid organelles, and that oligomerization but not IDRs of the EBOV nucleoprotein plays a key role in their formation. Additionally, VP35 (often considered the phosphoprotein-equivalent of EBOV) is not essential for IB formation, but alters their liquid behaviour. These findings define the molecular mechanism for the formation of EBOV IBs, which play a central role in the life cycle of this deadly virus.https://www.tandfonline.com/doi/10.1080/22221751.2023.2223727Ebolavirus, inclusion bodies, intrinsically disordered region, liquid organelle, liquid–liquid phase separation
spellingShingle Bianca S. Bodmer
Melina Vallbracht
Dmitry S. Ushakov
Lisa Wendt
Petr Chlanda
Thomas Hoenen
Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
Emerging Microbes and Infections
Ebolavirus, inclusion bodies, intrinsically disordered region, liquid organelle, liquid–liquid phase separation
title Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
title_full Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
title_fullStr Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
title_full_unstemmed Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
title_short Ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
title_sort ebola virus inclusion bodies are liquid organelles whose formation is facilitated by nucleoprotein oligomerization
topic Ebolavirus, inclusion bodies, intrinsically disordered region, liquid organelle, liquid–liquid phase separation
url https://www.tandfonline.com/doi/10.1080/22221751.2023.2223727
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