Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells.
Identifieur interne : 000844 ( PubMed/Checkpoint ); précédent : 000843; suivant : 000845Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells.
Auteurs : Shutoku Matsuyama [Japon] ; Kazuya Shirato [Japon] ; Miyuki Kawase [Japon] ; Yutaka Terada [Japon] ; Kengo Kawachi [Japon] ; Shuetsu Fukushi [Japon] ; Wataru Kamitani [Japon]Source :
- Journal of virology [ 1098-5514 ] ; 2018.
Descripteurs français
- KwdFr :
- Animaux, Cathepsine B (antagonistes et inhibiteurs), Cathepsine B (génétique), Cathepsine B (métabolisme), Cathepsine L (antagonistes et inhibiteurs), Cathepsine L (génétique), Cathepsine L (métabolisme), Cellules Vero, Chlorométhyl cétones d'acides aminés (pharmacologie), Coronavirus du syndrome respiratoire du Moyen-Orient (génétique), Coronavirus du syndrome respiratoire du Moyen-Orient (métabolisme), Furine (antagonistes et inhibiteurs), Furine (génétique), Furine (métabolisme), Glycoprotéine de spicule des coronavirus (génétique), Glycoprotéine de spicule des coronavirus (métabolisme), Humains, Papaïne (antagonistes et inhibiteurs), Papaïne (génétique), Papaïne (métabolisme), Protéolyse, Pénétration virale, Serine endopeptidases (génétique), Serine endopeptidases (métabolisme).
- MESH :
- antagonistes et inhibiteurs : Cathepsine B, Cathepsine L, Furine, Papaïne.
- génétique : Cathepsine B, Cathepsine L, Coronavirus du syndrome respiratoire du Moyen-Orient, Furine, Glycoprotéine de spicule des coronavirus, Papaïne, Serine endopeptidases.
- métabolisme : Cathepsine B, Cathepsine L, Coronavirus du syndrome respiratoire du Moyen-Orient, Furine, Glycoprotéine de spicule des coronavirus, Papaïne, Serine endopeptidases.
- pharmacologie : Chlorométhyl cétones d'acides aminés.
- Animaux, Cellules Vero, Humains, Protéolyse, Pénétration virale.
English descriptors
- KwdEn :
- Amino Acid Chloromethyl Ketones (pharmacology), Animals, Cathepsin B (antagonists & inhibitors), Cathepsin B (genetics), Cathepsin B (metabolism), Cathepsin L (antagonists & inhibitors), Cathepsin L (genetics), Cathepsin L (metabolism), Chlorocebus aethiops, Furin (antagonists & inhibitors), Furin (genetics), Furin (metabolism), Humans, Middle East Respiratory Syndrome Coronavirus (genetics), Middle East Respiratory Syndrome Coronavirus (metabolism), Papain (antagonists & inhibitors), Papain (genetics), Papain (metabolism), Proteolysis, Serine Endopeptidases (genetics), Serine Endopeptidases (metabolism), Spike Glycoprotein, Coronavirus (genetics), Spike Glycoprotein, Coronavirus (metabolism), Vero Cells, Virus Internalization.
- MESH :
- chemical , antagonists & inhibitors : Cathepsin B, Cathepsin L, Furin, Papain.
- chemical , genetics : Cathepsin B, Cathepsin L, Furin, Papain, Serine Endopeptidases, Spike Glycoprotein, Coronavirus.
- chemical , metabolism : Cathepsin B, Cathepsin L, Furin, Papain, Serine Endopeptidases, Spike Glycoprotein, Coronavirus.
- chemical , pharmacology : Amino Acid Chloromethyl Ketones.
- genetics : Middle East Respiratory Syndrome Coronavirus.
- metabolism : Middle East Respiratory Syndrome Coronavirus.
- Animals, Chlorocebus aethiops, Humans, Proteolysis, Vero Cells, Virus Internalization.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes host cellular proteases to enter cells. A previous report shows that furin, which is distributed mainly in the Golgi apparatus and cycled to the cell surface and endosomes, proteolytically activates the MERS-CoV spike (S) protein following receptor binding to mediate fusion between the viral and cellular membranes. In this study, we reexamined furin usage by MERS-CoV using a real-time PCR-based virus cell entry assay after inhibition of cellular proteases. We found that the furin inhibitor dec-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, dec-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2. Furthermore, a virus cell entry assay and a cell-cell fusion assay provided no evidence that the S protein was activated by exogenous furin. Therefore, we conclude that furin does not play a role in entry of MERS-CoV into cells and that the inhibitory effect of dec-RVKR-CMK is specific for TMPRSS2 and cathepsin L rather than furin.IMPORTANCE Previous studies using the furin inhibitor dec-RVKR-CMK suggest that MERS-CoV utilizes a cellular protease, furin, to activate viral glycoproteins during cell entry. However, we found that dec-RVKR-CMK inhibits not only furin but also other proteases. Furthermore, we found no evidence that MERS-CoV uses furin. These findings suggest that previous studies in the virology field based on dec-RVKR-CMK should be reexamined carefully. Here we describe appropriate experiments that can be used to assess the effect of protease inhibitors on virus cell entry.
DOI: 10.1128/JVI.00683-18
PubMed: 30021905
Affiliations:
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pubmed:30021905Le document en format XML
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<term>Cathepsin B (antagonists & inhibitors)</term>
<term>Cathepsin B (genetics)</term>
<term>Cathepsin B (metabolism)</term>
<term>Cathepsin L (antagonists & inhibitors)</term>
<term>Cathepsin L (genetics)</term>
<term>Cathepsin L (metabolism)</term>
<term>Chlorocebus aethiops</term>
<term>Furin (antagonists & inhibitors)</term>
<term>Furin (genetics)</term>
<term>Furin (metabolism)</term>
<term>Humans</term>
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<term>Middle East Respiratory Syndrome Coronavirus (metabolism)</term>
<term>Papain (antagonists & inhibitors)</term>
<term>Papain (genetics)</term>
<term>Papain (metabolism)</term>
<term>Proteolysis</term>
<term>Serine Endopeptidases (genetics)</term>
<term>Serine Endopeptidases (metabolism)</term>
<term>Spike Glycoprotein, Coronavirus (genetics)</term>
<term>Spike Glycoprotein, Coronavirus (metabolism)</term>
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<term>Cathepsine L (métabolisme)</term>
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<term>Chlorométhyl cétones d'acides aminés (pharmacologie)</term>
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<term>Furine (génétique)</term>
<term>Furine (métabolisme)</term>
<term>Glycoprotéine de spicule des coronavirus (génétique)</term>
<term>Glycoprotéine de spicule des coronavirus (métabolisme)</term>
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<term>Papaïne (antagonistes et inhibiteurs)</term>
<term>Papaïne (génétique)</term>
<term>Papaïne (métabolisme)</term>
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<term>Pénétration virale</term>
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<term>Coronavirus du syndrome respiratoire du Moyen-Orient</term>
<term>Furine</term>
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<term>Papaïne</term>
<term>Serine endopeptidases</term>
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<front><div type="abstract" xml:lang="en">Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes host cellular proteases to enter cells. A previous report shows that furin, which is distributed mainly in the Golgi apparatus and cycled to the cell surface and endosomes, proteolytically activates the MERS-CoV spike (S) protein following receptor binding to mediate fusion between the viral and cellular membranes. In this study, we reexamined furin usage by MERS-CoV using a real-time PCR-based virus cell entry assay after inhibition of cellular proteases. We found that the furin inhibitor dec-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, dec-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2. Furthermore, a virus cell entry assay and a cell-cell fusion assay provided no evidence that the S protein was activated by exogenous furin. Therefore, we conclude that furin does not play a role in entry of MERS-CoV into cells and that the inhibitory effect of dec-RVKR-CMK is specific for TMPRSS2 and cathepsin L rather than furin.<b>IMPORTANCE</b>
Previous studies using the furin inhibitor dec-RVKR-CMK suggest that MERS-CoV utilizes a cellular protease, furin, to activate viral glycoproteins during cell entry. However, we found that dec-RVKR-CMK inhibits not only furin but also other proteases. Furthermore, we found no evidence that MERS-CoV uses furin. These findings suggest that previous studies in the virology field based on dec-RVKR-CMK should be reexamined carefully. Here we describe appropriate experiments that can be used to assess the effect of protease inhibitors on virus cell entry.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30021905</PMID>
<DateCompleted><Year>2018</Year>
<Month>09</Month>
<Day>18</Day>
</DateCompleted>
<DateRevised><Year>2019</Year>
<Month>12</Month>
<Day>10</Day>
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<Issue>19</Issue>
<PubDate><Year>2018</Year>
<Month>10</Month>
<Day>01</Day>
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<Title>Journal of virology</Title>
<ISOAbbreviation>J. Virol.</ISOAbbreviation>
</Journal>
<ArticleTitle>Middle East Respiratory Syndrome Coronavirus Spike Protein Is Not Activated Directly by Cellular Furin during Viral Entry into Target Cells.</ArticleTitle>
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<Abstract><AbstractText>Middle East respiratory syndrome coronavirus (MERS-CoV) utilizes host cellular proteases to enter cells. A previous report shows that furin, which is distributed mainly in the Golgi apparatus and cycled to the cell surface and endosomes, proteolytically activates the MERS-CoV spike (S) protein following receptor binding to mediate fusion between the viral and cellular membranes. In this study, we reexamined furin usage by MERS-CoV using a real-time PCR-based virus cell entry assay after inhibition of cellular proteases. We found that the furin inhibitor dec-RVKR-CMK blocked entry of MERS-CoV harboring an S protein lacking furin cleavage sites; it even blocked entry into furin-deficient LoVo cells. In addition, dec-RVKR-CMK inhibited not only the enzymatic activity of furin but also those of cathepsin L, cathepsin B, trypsin, papain, and TMPRSS2. Furthermore, a virus cell entry assay and a cell-cell fusion assay provided no evidence that the S protein was activated by exogenous furin. Therefore, we conclude that furin does not play a role in entry of MERS-CoV into cells and that the inhibitory effect of dec-RVKR-CMK is specific for TMPRSS2 and cathepsin L rather than furin.<b>IMPORTANCE</b>
Previous studies using the furin inhibitor dec-RVKR-CMK suggest that MERS-CoV utilizes a cellular protease, furin, to activate viral glycoproteins during cell entry. However, we found that dec-RVKR-CMK inhibits not only furin but also other proteases. Furthermore, we found no evidence that MERS-CoV uses furin. These findings suggest that previous studies in the virology field based on dec-RVKR-CMK should be reexamined carefully. Here we describe appropriate experiments that can be used to assess the effect of protease inhibitors on virus cell entry.</AbstractText>
<CopyrightInformation>Copyright © 2018 American Society for Microbiology.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Matsuyama</LastName>
<ForeName>Shutoku</ForeName>
<Initials>S</Initials>
<AffiliationInfo><Affiliation>Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan matuyama@nih.go.jp.</Affiliation>
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<Author ValidYN="Y"><LastName>Shirato</LastName>
<ForeName>Kazuya</ForeName>
<Initials>K</Initials>
<AffiliationInfo><Affiliation>Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Kawase</LastName>
<ForeName>Miyuki</ForeName>
<Initials>M</Initials>
<AffiliationInfo><Affiliation>Department of Virology III, National Institute of Infectious Diseases, Tokyo, Japan.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Terada</LastName>
<ForeName>Yutaka</ForeName>
<Initials>Y</Initials>
<AffiliationInfo><Affiliation>Laboratory of Clinical Research on Infectious Diseases, Osaka University, Osaka, Japan.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Kawachi</LastName>
<ForeName>Kengo</ForeName>
<Initials>K</Initials>
<AffiliationInfo><Affiliation>Laboratory of Clinical Research on Infectious Diseases, Osaka University, Osaka, Japan.</Affiliation>
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