Live‐Cell Imaging of Ebola Virus Matrix Protein VP40

Frick, Cary T.; Ollmann-Saphire, Erica; Stahelin, Robert V.

doi:10.1096/fasebj.29.1_supplement.886.4

Cited by 2 publications

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“…Real-time monitoring systems for the intracellular dynamics of VP40 ( Adu-Gyamfi et al, 2012 ; Frick et al, 2015 ) and nucleocapsid-like structural proteins, including NP, VP35, and VP24 ( Takamatsu et al, 2018 ), in live cells have been developed. Integration with our system enables a more detailed characterization of molecular dynamics in terms of host–virus interactions during the EBOV replication cycle.…”

Section: Discussionmentioning

confidence: 99%

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

Furuyama

Sakaguchi²,

Yamada³

et al. 2022

Front. Microbiol.

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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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Section: Discussionmentioning

confidence: 99%

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

Furuyama

Sakaguchi²,

Yamada³

et al. 2022

Front. Microbiol.

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“…The EBOV structural protein VP40 is necessary and sufficient for the assembly and budding of virus particles. When expressed in the absence of any other viral protein, VP40 can form VLPs that bud and are released from cells similar to wild-type EBOV [46][47][48]. To determine if HERC5 targets VP40, we co-transfected 293T cells with a plasmid carrying VP40 and increasing concentrations of plasmids carrying either empty vector control or FLAG-tagged HERC5.…”

Section: Herc5 Inhibits Ebov Vp40 Particle Productionmentioning

confidence: 99%

Interferon-Induced HERC5 Inhibits Ebola Virus Particle Production and Is Antagonized by Ebola Glycoprotein

Paparisto

Hunt

Labach

et al. 2021

Cells

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Survival following Ebola virus (EBOV) infection correlates with the ability to mount an early and robust interferon (IFN) response. The host IFN-induced proteins that contribute to controlling EBOV replication are not fully known. Among the top genes with the strongest early increases in expression after infection in vivo is IFN-induced HERC5. Using a transcription- and replication-competent VLP system, we showed that HERC5 inhibits EBOV virus-like particle (VLP) replication by depleting EBOV mRNAs. The HERC5 RCC1-like domain was necessary and sufficient for this inhibition and did not require zinc finger antiviral protein (ZAP). Moreover, we showed that EBOV (Zaire) glycoprotein (GP) but not Marburg virus GP antagonized HERC5 early during infection. Our data identify a novel ‘protagonist–antagonistic’ relationship between HERC5 and GP in the early stages of EBOV infection that could be exploited for the development of novel antiviral therapeutics.

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Live‐Cell Imaging of Ebola Virus Matrix Protein VP40

Cited by 2 publications

References 0 publications

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

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

Interferon-Induced HERC5 Inhibits Ebola Virus Particle Production and Is Antagonized by Ebola Glycoprotein

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