Gating kinetics of pH-activated membrane fusion of vesicular stomatitis virus with cells: stopped-flow measurements by dequenching of octadecylrhodamine fluorescence

Biochemistry ◽  
1990 ◽  
Vol 29 (5) ◽  
pp. 1303-1308 ◽  
Author(s):  
Michael J. Clague ◽  
Christian Schoch ◽  
Loren Zech ◽  
Robert Blumenthal
Keyword(s):  
Membrane Fusion ◽  
Vesicular Stomatitis Virus ◽  
Stopped Flow ◽  
Flow Measurements ◽  
Gating Kinetics ◽  
Vesicular Stomatitis ◽  
Kinetics Of

scholarly journals Membrane Fusion Induced by Vesicular Stomatitis Virus Depends on Histidine Protonation

2003 ◽  
Vol 278 (16) ◽  
pp. 13789-13794 ◽  
Author(s):  
Fabiana A. Carneiro ◽  
Fausto Stauffer ◽  
Carla S. Lima ◽  
Maria A. Juliano ◽  
Luiz Juliano ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Vesicular Stomatitis Virus ◽  
Vesicular Stomatitis

scholarly journals The Membrane-Proximal Domain of Vesicular Stomatitis Virus G Protein Functions as a Membrane Fusion Potentiator and Can Induce Hemifusion

2002 ◽  
Vol 76 (23) ◽  
pp. 12300-12311 ◽  
Author(s):  
E. Jeetendra ◽  
Clinton S. Robison ◽  
Lorraine M. Albritton ◽  
Michael A. Whitt
Keyword(s):  
Membrane Fusion ◽  
G Protein ◽  
Vesicular Stomatitis Virus ◽  
Fusion Proteins ◽  
Fold Increase ◽  
Wild Type Virus ◽  
Viral Fusion ◽  
Fusion Activity ◽  
Viral Glycoprotein ◽  
Vesicular Stomatitis

ABSTRACT Recently we showed that the membrane-proximal stem region of the vesicular stomatitis virus (VSV) G protein ectodomain (G stem [GS]), together with the transmembrane and cytoplasmic domains, was sufficient to mediate efficient VSV budding (C. S. Robison and M. A. Whitt, J. Virol. 74:2239-2246, 2000). Here, we show that GS can also potentiate the membrane fusion activity of heterologous viral fusion proteins when GS is coexpressed with those proteins. For some fusion proteins, there was as much as a 40-fold increase in syncytium formation when GS was coexpressed compared to that seen when the fusion protein was expressed alone. Fusion potentiation by GS was not protein specific, since it occurred with both pH-dependent as well as pH-independent fusion proteins. Using a recombinant vesicular stomatitis virus encoding GS that contained an N-terminal hemagglutinin (HA) tag (GSHA virus), we found that the GSHA virus bound to cells as well as the wild-type virus did at pH 7.0; however, the GSHA virus was noninfectious. Analysis of cells expressing GSHA in a three-color membrane fusion assay revealed that GSHA could induce lipid mixing but not cytoplasmic mixing, indicating that GS can induce hemifusion. Treatment of GSHA virus-bound cells with the membrane-destabilizing drug chlorpromazine rescued the hemifusion block and allowed entry and subsequent replication of GSHA virus, demonstrating that GS-mediated hemifusion was a functional intermediate in the membrane fusion pathway. Using a series of truncation mutants, we also determined that only 14 residues of GS, together with the VSV G transmembrane and cytoplasmic tail, were sufficient for fusion potentiation. To our knowledge, this is the first report which shows that a small domain of one viral glycoprotein can promote the fusion activity of other, unrelated viral glycoproteins.

scholarly journals Cholesterol 25-hydroxylase suppresses SARS-CoV-2 replication by blocking membrane fusion

2020 ◽  
Vol 117 (50) ◽  
pp. 32105-32113 ◽  
Author(s):  
Ruochen Zang ◽  
James Brett Case ◽  
Eylan Yutuc ◽  
Xiucui Ma ◽  
Sheng Shen ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Vesicular Stomatitis Virus ◽  
Molecular Basis ◽  
Enveloped Viruses ◽  
Late Endosomes ◽  
Wide Range ◽  
Therapeutic Development ◽  
Antiviral Activities ◽  
Vesicular Stomatitis ◽  
Catalyzed Membrane

Cholesterol 25-hydroxylase (CH25H) is an interferon (IFN)-stimulated gene that shows broad antiviral activities against a wide range of enveloped viruses. Here, using an IFN-stimulated gene screen against vesicular stomatitis virus (VSV)-SARS-CoV and VSV-SARS-CoV-2 chimeric viruses, we identified CH25H and its enzymatic product 25-hydroxycholesterol (25HC) as potent inhibitors of SARS-CoV-2 replication. Internalized 25HC accumulates in the late endosomes and potentially restricts SARS-CoV-2 spike protein catalyzed membrane fusion via blockade of cholesterol export. Our results highlight one of the possible antiviral mechanisms of 25HC and provide the molecular basis for its therapeutic development.

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