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2.
Antiviral Res ; 200: 105270, 2022 04.
Article in English | MEDLINE | ID: covidwho-1763566

ABSTRACT

The pandemic caused by the new coronavirus SARS-CoV-2 has made evident the need for broad-spectrum, efficient antiviral treatments to combat emerging and re-emerging viruses. Plitidepsin is an antitumor agent of marine origin that has also shown a potent pre-clinical efficacy against SARS-CoV-2. Plitidepsin targets the host protein eEF1A (eukaryotic translation elongation factor 1 alpha) and affects viral infection at an early, post-entry step. Because electron microscopy is a valuable tool to study virus-cell interactions and the mechanism of action of antiviral drugs, in this work we have used transmission electron microscopy (TEM) to evaluate the effects of plitidepsin in SARS-CoV-2 infection in cultured Vero E6 cells 24 and 48h post-infection. In the absence of plitidepsin, TEM morphological analysis showed double-membrane vesicles (DMVs), organelles that support coronavirus genome replication, single-membrane vesicles with viral particles, large vacuoles with groups of viruses and numerous extracellular virions attached to the plasma membrane. When treated with plitidepsin, no viral structures were found in SARS-CoV-2-infected Vero E6 cells. Immunogold detection of SARS-CoV-2 nucleocapsid (N) protein and double-stranded RNA (dsRNA) provided clear signals in cells infected in the absence of plitidepsin, but complete absence in cells infected and treated with plitidepsin. The present study shows that plitidepsin blocks the biogenesis of viral replication organelles and the morphogenesis of virus progeny. Electron microscopy morphological analysis coupled to immunogold labeling of SARS-CoV-2 products offers a unique approach to understand how antivirals such as plitidepsin work.


Subject(s)
COVID-19 , Depsipeptides , Animals , Antiviral Agents/therapeutic use , COVID-19/drug therapy , Chlorocebus aethiops , Depsipeptides/pharmacology , Peptides, Cyclic , SARS-CoV-2 , Vero Cells , Virus Replication
3.
Viruses ; 13(10)2021 09 29.
Article in English | MEDLINE | ID: covidwho-1688586

ABSTRACT

Recent advances in light and electron microscopy are uncovering viral lifecycle events with a level of detail never before seen [...].


Subject(s)
Host Microbial Interactions , Image Interpretation, Computer-Assisted/methods , Single Molecule Imaging/methods , Humans , Image Interpretation, Computer-Assisted/instrumentation , Microscopy, Electron/methods , Single Molecule Imaging/instrumentation , Virus Replication
5.
Encyclopedia of Virology ; : 495-500, 2021.
Article in English | [Unspecified Source], Grey literature | ID: grc-750375

ABSTRACT

Viral factories are intracellular compartments of the host cell that contain viral replication organelles and necessary elements for assembly and maturation of new infectious viral particles. In this article we revise the methods used to study viral factories and the current knowledge on the structure, functions and biogenesis of these structures. We also describe some of the most emblematic examples of viral factories characterized so far. Finally, we describe how the identification of mechanisms involved in the biogenesis and functional architecture of viral factories will provide new means for antiviral intervention.

6.
Non-conventional | WHO COVID | ID: covidwho-634859

ABSTRACT

Viral factories are intracellular compartments of the host cell that contain viral replication organelles and necessary elements for assembly and maturation of new infectious viral particles. In this article we revise the methods used to study viral factories and the current knowledge on the structure, functions and biogenesis of these structures. We also describe some of the most emblematic examples of viral factories characterized so far. Finally, we describe how the identification of mechanisms involved in the biogenesis and functional architecture of viral factories will provide new means for antiviral intervention.

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