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J Physiol
2021 Jun 01;59911:2851-2868. doi: 10.1113/JP281037.
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The envelope protein of SARS-CoV-2 increases intra-Golgi pH and forms a cation channel that is regulated by pH.
Cabrera-Garcia D
,
Bekdash R
,
Abbott GW
,
Yazawa M
,
Harrison NL
.
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KEY POINTS: We report a novel method for the transient expression of SARS-CoV-2 envelope (E) protein in intracellular organelles and the plasma membrane of mammalian cells and Xenopus oocytes. Intracellular expression of SARS-CoV-2 E protein increases intra-Golgi pH. By targeting the SARS-CoV-2 E protein to the plasma membrane, we show that it forms a cation channel, viroporin, that is modulated by changes of pH. This method for studying the activity of viroporins may facilitate screening for new antiviral drugs to identify novel treatments for COVID-19.
ABSTRACT: The envelope (E) protein of coronaviruses such as SARS-CoV-1 is proposed to form an ion channel or viroporin that participates in viral propagation and pathogenesis. Here we developed a technique to study the E protein of SARS-CoV-2 in mammalian cells by directed targeting using a carboxyl-terminal fluorescent protein tag, mKate2. The wild-type SARS-CoV-2 E protein can be trafficked to intracellular organelles, notably the endoplasmic reticulum-Golgi intermediate complex, where its expression increases pH inside the organelle. We also succeeded in targeting SARS-CoV-2 E to the plasma membrane, which enabled biophysical analysis using whole-cell patch clamp recording in a mammalian cell line, HEK 293 cells, and two-electrode voltage clamp electrophysiology in Xenopus oocytes. The results suggest that the E protein forms an ion channel that is permeable to monovalent cations such as Na+ , Cs+ and K+ . The E current is nearly time- and voltage-independent when E protein is expressed in mammalian cells, and is modulated by changes of pH. At pH 6.0 and 7.4, the E protein current is activated, whereas at pH 8.0 and 9.0, the amplitude of E protein current is reduced, and in oocytes the inward E current fades at pH 9 in a time- and voltage-dependent manner. Using this directed targeting method and electrophysiological recordings, potential inhibitors of the E protein can be screened and subsequently investigated for antiviral activity against SARS-CoV-2 in vitro and possible efficacy in treating COVID-19.
Columbia Stem Cell Initiative, Department of Anesthesiology at CUIMC, Columbia University Irving Medical Centre Dean's Office Fund, GM130377 NIH, National Institute of General Medical Sciences, R35 GM130377 NIGMS NIH HHS
Abbott,
KCNE1 and KCNE3: The yin and yang of voltage-gated K(+) channel regulation.
2016, Pubmed
Abbott,
KCNE1 and KCNE3: The yin and yang of voltage-gated K(+) channel regulation.
2016,
Pubmed
Andersen,
The proximal origin of SARS-CoV-2.
2020,
Pubmed
Arons,
Presymptomatic SARS-CoV-2 Infections and Transmission in a Skilled Nursing Facility.
2020,
Pubmed
Bedford,
Cryptic transmission of SARS-CoV-2 in Washington state.
2020,
Pubmed
Bianchi,
Sars-CoV-2 Envelope and Membrane Proteins: Structural Differences Linked to Virus Characteristics?
2020,
Pubmed
Boson,
The SARS-CoV-2 envelope and membrane proteins modulate maturation and retention of the spike protein, allowing assembly of virus-like particles.
2021,
Pubmed
Castaño-Rodriguez,
Role of Severe Acute Respiratory Syndrome Coronavirus Viroporins E, 3a, and 8a in Replication and Pathogenesis.
2018,
Pubmed
Cele,
Escape of SARS-CoV-2 501Y.V2 from neutralization by convalescent plasma.
2021,
Pubmed
Chizhmakov,
Selective proton permeability and pH regulation of the influenza virus M2 channel expressed in mouse erythroleukaemia cells.
1996,
Pubmed
Cohen,
Identification of a Golgi complex-targeting signal in the cytoplasmic tail of the severe acute respiratory syndrome coronavirus envelope protein.
2011,
Pubmed
Corse,
The cytoplasmic tails of infectious bronchitis virus E and M proteins mediate their interaction.
2003,
Pubmed
Cummings,
Epidemiology, clinical course, and outcomes of critically ill adults with COVID-19 in New York City: a prospective cohort study.
2020,
Pubmed
DeDiego,
A severe acute respiratory syndrome coronavirus that lacks the E gene is attenuated in vitro and in vivo.
2007,
Pubmed
DeDiego,
Severe acute respiratory syndrome coronavirus envelope protein regulates cell stress response and apoptosis.
2011,
Pubmed
do Espírito Santo,
In vivo demonstration of microvascular thrombosis in severe COVID-19.
2020,
Pubmed
Dolin,
A controlled trial of amantadine and rimantadine in the prophylaxis of influenza A infection.
1982,
Pubmed
Duart,
SARS-CoV-2 envelope protein topology in eukaryotic membranes.
2020,
Pubmed
Farag,
Viroporins and inflammasomes: A key to understand virus-induced inflammation.
2020,
Pubmed
Giannis,
Coagulation disorders in coronavirus infected patients: COVID-19, SARS-CoV-1, MERS-CoV and lessons from the past.
2020,
Pubmed
Gonzalez-Reiche,
Introductions and early spread of SARS-CoV-2 in the New York City area.
2020,
Pubmed
Griffin,
The p7 protein of hepatitis C virus forms an ion channel that is blocked by the antiviral drug, Amantadine.
2003,
Pubmed
Gupta,
In-silico approaches to detect inhibitors of the human severe acute respiratory syndrome coronavirus envelope protein ion channel.
2021,
Pubmed
Hamill,
Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.
1981,
Pubmed
Helenius,
Unpacking the incoming influenza virus.
1992,
Pubmed
Hofherr,
Selective Golgi export of Kir2.1 controls the stoichiometry of functional Kir2.x channel heteromers.
2005,
Pubmed
Huang,
Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: implications for assembly and vaccine production.
2004,
Pubmed
Jiang,
Endogenous Kv channels in human embryonic kidney (HEK-293) cells.
2002,
Pubmed
Jimenez-Guardeño,
The PDZ-binding motif of severe acute respiratory syndrome coronavirus envelope protein is a determinant of viral pathogenesis.
2014,
Pubmed
Klok,
Incidence of thrombotic complications in critically ill ICU patients with COVID-19.
2020,
Pubmed
Li,
Angiotensin-converting enzyme 2 is a functional receptor for the SARS coronavirus.
2003,
Pubmed
Lim,
The missing link in coronavirus assembly. Retention of the avian coronavirus infectious bronchitis virus envelope protein in the pre-Golgi compartments and physical interaction between the envelope and membrane proteins.
2001,
Pubmed
Mandala,
Structure and drug binding of the SARS-CoV-2 envelope protein transmembrane domain in lipid bilayers.
2020,
Pubmed
Matamala,
Imaging the electrical activity of organelles in living cells.
2021,
Pubmed
McClenaghan,
Coronavirus Proteins as Ion Channels: Current and Potential Research.
2020,
Pubmed
Mehta,
COVID-19: consider cytokine storm syndromes and immunosuppression.
2020,
Pubmed
Moghadas,
The implications of silent transmission for the control of COVID-19 outbreaks.
2020,
Pubmed
Nieto-Torres,
Subcellular location and topology of severe acute respiratory syndrome coronavirus envelope protein.
2011,
Pubmed
Nieto-Torres,
Severe acute respiratory syndrome coronavirus envelope protein ion channel activity promotes virus fitness and pathogenesis.
2014,
Pubmed
Nieto-Torres,
Relevance of Viroporin Ion Channel Activity on Viral Replication and Pathogenesis.
2015,
Pubmed
Nieto-Torres,
Severe acute respiratory syndrome coronavirus E protein transports calcium ions and activates the NLRP3 inflammasome.
2015,
Pubmed
Nieva,
Viroporins: structure and biological functions.
2012,
Pubmed
Pei,
Differential effects of intervention timing on COVID-19 spread in the United States.
2020,
Pubmed
Pervushin,
Structure and inhibition of the SARS coronavirus envelope protein ion channel.
2009,
Pubmed
Pinto,
Influenza virus M2 protein has ion channel activity.
1992,
Pubmed
,
Xenbase
Premkumar,
Cation-selective ion channels formed by p7 of hepatitis C virus are blocked by hexamethylene amiloride.
2004,
Pubmed
Regla-Nava,
The replication of a mouse adapted SARS-CoV in a mouse cell line stably expressing the murine SARS-CoV receptor mACE2 efficiently induces the expression of proinflammatory cytokines.
2013,
Pubmed
Schnell,
Structure and mechanism of the M2 proton channel of influenza A virus.
2008,
Pubmed
Schoeman,
Coronavirus envelope protein: current knowledge.
2019,
Pubmed
Schoeman,
Is There a Link Between the Pathogenic Human Coronavirus Envelope Protein and Immunopathology? A Review of the Literature.
2020,
Pubmed
Scott,
Viroporins: structure, function and potential as antiviral targets.
2015,
Pubmed
Shang,
Cell entry mechanisms of SARS-CoV-2.
2020,
Pubmed
Shcherbo,
Far-red fluorescent tags for protein imaging in living tissues.
2009,
Pubmed
,
Xenbase
Singh Tomar,
SARS-CoV-2 E protein is a potential ion channel that can be inhibited by Gliclazide and Memantine.
2020,
Pubmed
Stadlbauer,
Repeated cross-sectional sero-monitoring of SARS-CoV-2 in New York City.
2021,
Pubmed
Torres,
Conductance and amantadine binding of a pore formed by a lysine-flanked transmembrane domain of SARS coronavirus envelope protein.
2007,
Pubmed
Torres,
Model of a putative pore: the pentameric alpha-helical bundle of SARS coronavirus E protein in lipid bilayers.
2006,
Pubmed
Vaughan,
Exhaled breath condensate pH is a robust and reproducible assay of airway acidity.
2003,
Pubmed
Venkatagopalan,
Coronavirus envelope (E) protein remains at the site of assembly.
2015,
Pubmed
Wang,
Serological Evidence of Bat SARS-Related Coronavirus Infection in Humans, China.
2018,
Pubmed
Washington,
Emergence and rapid transmission of SARS-CoV-2 B.1.1.7 in the United States.
2021,
Pubmed
Westerbeck,
The Infectious Bronchitis Coronavirus Envelope Protein Alters Golgi pH To Protect the Spike Protein and Promote the Release of Infectious Virus.
2019,
Pubmed
Wibmer,
SARS-CoV-2 501Y.V2 escapes neutralization by South African COVID-19 donor plasma.
2021,
Pubmed
Williamson,
Factors associated with COVID-19-related death using OpenSAFELY.
2020,
Pubmed
Wilson,
SARS coronavirus E protein forms cation-selective ion channels.
2004,
Pubmed
Wilson,
Hexamethylene amiloride blocks E protein ion channels and inhibits coronavirus replication.
2006,
Pubmed
Worobey,
The emergence of SARS-CoV-2 in Europe and North America.
2020,
Pubmed
Wozniak,
Intracellular proton conductance of the hepatitis C virus p7 protein and its contribution to infectious virus production.
2010,
Pubmed
Xia,
SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target.
2021,
Pubmed
Yue,
SARS-Coronavirus Open Reading Frame-3a drives multimodal necrotic cell death.
2018,
Pubmed
Zhang,
Histopathologic Changes and SARS-CoV-2 Immunostaining in the Lung of a Patient With COVID-19.
2020,
Pubmed
Zhou,
Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan, China: a retrospective cohort study.
2020,
Pubmed
Zhou,
A pneumonia outbreak associated with a new coronavirus of probable bat origin.
2020,
Pubmed
Zhu,
A Novel Coronavirus from Patients with Pneumonia in China, 2019.
2020,
Pubmed
