Monitoring of S Protein Maturation in the Endoplasmic Reticulum by Calnexin Is Important for the Infectivity of Severe Acute Respiratory Syndrome Coronavirus
Identifieur interne : 001F73 ( Main/Exploration ); précédent : 001F72; suivant : 001F74Monitoring of S Protein Maturation in the Endoplasmic Reticulum by Calnexin Is Important for the Infectivity of Severe Acute Respiratory Syndrome Coronavirus
Auteurs : Masaya Fukushi [Japon] ; Yoshiyuki Yoshinaka [Japon] ; Yusuke Matsuoka [Japon] ; Seisuke Hatakeyama [Japon] ; Yukihito Ishizaka [Japon] ; Teruo Kirikae [Japon] ; Takehiko Sasazuki [Japon] ; Tohru Miyoshi-Akiyama [Japon]Source :
- Journal of virology [ 0022-538X ] ; 2012.
Descripteurs français
- KwdFr :
- Animaux, Calnexine (métabolisme), Glycoprotéine de spicule des coronavirus, Glycoprotéines membranaires (métabolisme), Glycosylation, Humains, Liaison aux protéines, Lignée cellulaire, Maturation post-traductionnelle des protéines, Protéines de l'enveloppe virale (métabolisme), Réplication virale, Réticulum endoplasmique (métabolisme), Souris, Virus du SRAS (pathogénicité), Virus du SRAS (physiologie).
- MESH :
- métabolisme : Calnexine, Glycoprotéines membranaires, Protéines de l'enveloppe virale, Réticulum endoplasmique.
- pathogénicité : Virus du SRAS.
- physiologie : Virus du SRAS.
- Pascal (Inist)
English descriptors
- KwdEn :
- Animals, Calnexin (metabolism), Cell Line, Coronavirus, Endoplasmic Reticulum (metabolism), Endoplasmic reticulum, Glycosylation, Humans, Infectivity, Membrane Glycoproteins (metabolism), Mice, Protein, Protein Binding, Protein Processing, Post-Translational, SARS Virus (pathogenicity), SARS Virus (physiology), Severe acute respiratory syndrome, Spike Glycoprotein, Coronavirus, Viral Envelope Proteins (metabolism), Virus Replication.
- MESH :
- chemical , metabolism : Calnexin, Membrane Glycoproteins, Viral Envelope Proteins.
- metabolism : Endoplasmic Reticulum.
- pathogenicity : SARS Virus.
- physiology : SARS Virus.
- Animals, Cell Line, Glycosylation, Humans, Mice, Protein Binding, Protein Processing, Post-Translational, Spike Glycoprotein, Coronavirus, Virus Replication.
Abstract
Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of SARS, a fatal pulmonary disorder with no effective treatment. We found that SARS-CoV spike glycoprotein (S protein), a key molecule for viral entry, binds to calnexin, a molecular chaperone in the endoplasmic reticulum (ER), but not to calreticulin, a homolog of calnexin. Calnexin bound to most truncated mutants of S protein, and S protein bound to all mutants of calnexin. Pseudotyped virus carrying S protein (S-pseudovirus) produced by human cells that were treated with small interfering RNA (siRNA) for calnexin expression (calnexin siRNA-treated cells) showed significantly lower infectivity than S-pseudoviruses produced by untreated and control siRNA-treated cells. S-pseudovirus produced by calnexin siRNA-treated cells contained S protein modified with N-glycan side chains differently from other two S proteins and consisted of two kinds of viral particles: those of normal density with little S protein and those of high density with abundant S protein. Treatment with peptide-N-glycosidase F (PNGase F), which removes all types of N-glycan side chains from glycoproteins, eliminated the infectivity of S-pseudovirus. S-pseudovirus and SARS-CoV produced in the presence of α-glucosidase inhibitors, which disrupt the interaction between calnexin and its substrates, showed significantly lower infectivity than each virus produced in the absence of those compounds. In S-pseudovirus, the incorporation of S protein into viral particles was obviously inhibited. In SARS-CoV, viral production was obviously inhibited. These findings demonstrated that calnexin strictly monitors the maturation of S protein by its direct binding, resulting in conferring infectivity on SARS-CoV.
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Infectivity of Severe Acute Respiratory Syndrome Coronavirus</title><author><name sortKey="Fukushi, Masaya" sort="Fukushi, Masaya" uniqKey="Fukushi M" first="Masaya" last="Fukushi">Masaya Fukushi</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation><affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Disease Control and Prevention Center, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation><affiliation wicri:level="3"><inist:fA14 i1="03"><s1>Deputy Director-General's Laboratory, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>Department of Virology, Institute of Biomedical & Health Sciences, Hiroshima University</s1><s2>Hiroshima</s2><s3>JPN</s3><sZ>1 aut.</sZ></inist:fA14><country>Japon</country><wicri:noRegion>Hiroshima</wicri:noRegion></affiliation></author><author><name sortKey="Yoshinaka, Yoshiyuki" sort="Yoshinaka, Yoshiyuki" uniqKey="Yoshinaka Y" first="Yoshiyuki" last="Yoshinaka">Yoshiyuki Yoshinaka</name><affiliation wicri:level="3"><inist:fA14 i1="05"><s1>Department of Molecular Virology, Bio-Response, Graduate School, Tokyo Medical and Dental University</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>2 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Matsuoka, Yusuke" sort="Matsuoka, Yusuke" uniqKey="Matsuoka Y" first="Yusuke" last="Matsuoka">Yusuke Matsuoka</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Hatakeyama, Seisuke" sort="Hatakeyama, Seisuke" uniqKey="Hatakeyama S" first="Seisuke" last="Hatakeyama">Seisuke Hatakeyama</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Ishizaka, Yukihito" sort="Ishizaka, Yukihito" uniqKey="Ishizaka Y" first="Yukihito" last="Ishizaka">Yukihito Ishizaka</name><affiliation wicri:level="3"><inist:fA14 i1="06"><s1>Department of Intractable Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>5 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Kirikae, Teruo" sort="Kirikae, Teruo" uniqKey="Kirikae T" first="Teruo" last="Kirikae">Teruo Kirikae</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Sasazuki, Takehiko" sort="Sasazuki, Takehiko" uniqKey="Sasazuki T" first="Takehiko" last="Sasazuki">Takehiko Sasazuki</name><affiliation wicri:level="3"><inist:fA14 i1="07"><s1>National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>7 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author><author><name sortKey="Miyoshi Akiyama, Tohru" sort="Miyoshi Akiyama, Tohru" uniqKey="Miyoshi Akiyama T" first="Tohru" last="Miyoshi-Akiyama">Tohru Miyoshi-Akiyama</name><affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Department of Infectious Diseases, Research Institute, National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>1 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>6 aut.</sZ><sZ>8 aut.</sZ></inist:fA14><country>Japon</country><placeName><settlement type="city">Tokyo</settlement><region type="région">Région de Kantō</region></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno><imprint><date when="2012">2012</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Calnexin (metabolism)</term><term>Cell Line</term><term>Coronavirus</term><term>Endoplasmic Reticulum (metabolism)</term><term>Endoplasmic reticulum</term><term>Glycosylation</term><term>Humans</term><term>Infectivity</term><term>Membrane Glycoproteins (metabolism)</term><term>Mice</term><term>Protein</term><term>Protein Binding</term><term>Protein Processing, Post-Translational</term><term>SARS Virus (pathogenicity)</term><term>SARS Virus (physiology)</term><term>Severe acute respiratory syndrome</term><term>Spike Glycoprotein, Coronavirus</term><term>Viral Envelope Proteins (metabolism)</term><term>Virus Replication</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Calnexine (métabolisme)</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Glycosylation</term><term>Humains</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term><term>Maturation post-traductionnelle des protéines</term><term>Protéines de l'enveloppe virale (métabolisme)</term><term>Réplication virale</term><term>Réticulum endoplasmique (métabolisme)</term><term>Souris</term><term>Virus du SRAS (pathogénicité)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Calnexin</term><term>Membrane Glycoproteins</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Endoplasmic Reticulum</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Calnexine</term><term>Glycoprotéines membranaires</term><term>Protéines de l'enveloppe virale</term><term>Réticulum endoplasmique</term></keywords><keywords scheme="MESH" qualifier="pathogenicity" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="pathogénicité" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cell Line</term><term>Glycosylation</term><term>Humans</term><term>Mice</term><term>Protein Binding</term><term>Protein Processing, Post-Translational</term><term>Spike Glycoprotein, Coronavirus</term><term>Virus Replication</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term><term>Coronavirus</term><term>Glycoprotéine de spicule des coronavirus</term><term>Glycosylation</term><term>Humains</term><term>Liaison aux protéines</term><term>Lignée cellulaire</term><term>Maturation post-traductionnelle des protéines</term><term>Protéine</term><term>Réplication virale</term><term>Réticulum endoplasmique</term><term>Pouvoir infectant</term><term>Souris</term><term>Syndrome respiratoire aigu sévère</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Severe acute respiratory syndrome coronavirus (SARS-CoV) is the etiological agent of SARS, a fatal pulmonary disorder with no effective treatment. We found that SARS-CoV spike glycoprotein (S protein), a key molecule for viral entry, binds to calnexin, a molecular chaperone in the endoplasmic reticulum (ER), but not to calreticulin, a homolog of calnexin. Calnexin bound to most truncated mutants of S protein, and S protein bound to all mutants of calnexin. Pseudotyped virus carrying S protein (S-pseudovirus) produced by human cells that were treated with small interfering RNA (siRNA) for calnexin expression (calnexin siRNA-treated cells) showed significantly lower infectivity than S-pseudoviruses produced by untreated and control siRNA-treated cells. S-pseudovirus produced by calnexin siRNA-treated cells contained S protein modified with N-glycan side chains differently from other two S proteins and consisted of two kinds of viral particles: those of normal density with little S protein and those of high density with abundant S protein. Treatment with peptide-N-glycosidase F (PNGase F), which removes all types of N-glycan side chains from glycoproteins, eliminated the infectivity of S-pseudovirus. S-pseudovirus and SARS-CoV produced in the presence of α-glucosidase inhibitors, which disrupt the interaction between calnexin and its substrates, showed significantly lower infectivity than each virus produced in the absence of those compounds. In S-pseudovirus, the incorporation of S protein into viral particles was obviously inhibited. In SARS-CoV, viral production was obviously inhibited. These findings demonstrated that calnexin strictly monitors the maturation of S protein by its direct binding, resulting in conferring infectivity on SARS-CoV.</div></front></TEI><affiliations><list><country><li>Japon</li></country><region><li>Région de Kantō</li></region><settlement><li>Tokyo</li></settlement></list><tree><country name="Japon"><region name="Région de Kantō"><name sortKey="Fukushi, Masaya" sort="Fukushi, Masaya" uniqKey="Fukushi M" first="Masaya" last="Fukushi">Masaya Fukushi</name></region><name sortKey="Fukushi, Masaya" sort="Fukushi, Masaya" uniqKey="Fukushi M" first="Masaya" last="Fukushi">Masaya Fukushi</name><name sortKey="Fukushi, Masaya" sort="Fukushi, Masaya" uniqKey="Fukushi M" first="Masaya" last="Fukushi">Masaya Fukushi</name><name sortKey="Fukushi, Masaya" sort="Fukushi, Masaya" uniqKey="Fukushi M" first="Masaya" last="Fukushi">Masaya Fukushi</name><name sortKey="Hatakeyama, Seisuke" sort="Hatakeyama, Seisuke" uniqKey="Hatakeyama S" first="Seisuke" last="Hatakeyama">Seisuke Hatakeyama</name><name sortKey="Ishizaka, Yukihito" sort="Ishizaka, Yukihito" uniqKey="Ishizaka Y" first="Yukihito" last="Ishizaka">Yukihito Ishizaka</name><name sortKey="Kirikae, Teruo" sort="Kirikae, Teruo" uniqKey="Kirikae T" first="Teruo" last="Kirikae">Teruo Kirikae</name><name sortKey="Matsuoka, Yusuke" sort="Matsuoka, Yusuke" uniqKey="Matsuoka Y" first="Yusuke" last="Matsuoka">Yusuke Matsuoka</name><name sortKey="Miyoshi Akiyama, Tohru" sort="Miyoshi Akiyama, Tohru" uniqKey="Miyoshi Akiyama T" first="Tohru" last="Miyoshi-Akiyama">Tohru Miyoshi-Akiyama</name><name sortKey="Sasazuki, Takehiko" sort="Sasazuki, Takehiko" uniqKey="Sasazuki T" first="Takehiko" last="Sasazuki">Takehiko Sasazuki</name><name sortKey="Yoshinaka, Yoshiyuki" sort="Yoshinaka, Yoshiyuki" uniqKey="Yoshinaka Y" first="Yoshiyuki" last="Yoshinaka">Yoshiyuki Yoshinaka</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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