Monitoring of S Protein Maturation in the Endoplasmic Reticulum by Calnexin Is Important for the Infectivity of Severe Acute Respiratory Syndrome Coronavirus
Identifieur interne : 000046 ( PascalFrancis/Checkpoint ); précédent : 000045; suivant : 000047Monitoring 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.
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- Pascal (Inist)
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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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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 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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>Coronavirus</term><term>Endoplasmic reticulum</term><term>Infectivity</term><term>Protein</term><term>Severe acute respiratory syndrome</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Protéine</term><term>Réticulum endoplasmique</term><term>Pouvoir infectant</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><inist><standard h6="B"><pA><fA01 i1="01" i2="1"><s0>0022-538X</s0></fA01><fA03 i2="1"><s0>J. virol.</s0></fA03><fA05><s2>86</s2></fA05><fA06><s2>21</s2></fA06><fA08 i1="01" i2="1" l="ENG"><s1>Monitoring of S Protein Maturation in the Endoplasmic Reticulum by Calnexin Is Important for the Infectivity of Severe Acute Respiratory Syndrome Coronavirus</s1></fA08><fA11 i1="01" i2="1"><s1>FUKUSHI (Masaya)</s1></fA11><fA11 i1="02" i2="1"><s1>YOSHINAKA (Yoshiyuki)</s1></fA11><fA11 i1="03" i2="1"><s1>MATSUOKA (Yusuke)</s1></fA11><fA11 i1="04" i2="1"><s1>HATAKEYAMA (Seisuke)</s1></fA11><fA11 i1="05" i2="1"><s1>ISHIZAKA (Yukihito)</s1></fA11><fA11 i1="06" i2="1"><s1>KIRIKAE (Teruo)</s1></fA11><fA11 i1="07" i2="1"><s1>SASAZUKI (Takehiko)</s1></fA11><fA11 i1="08" i2="1"><s1>MIYOSHI-AKIYAMA (Tohru)</s1></fA11><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></fA14><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></fA14><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></fA14><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></fA14><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></fA14><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></fA14><fA14 i1="07"><s1>National Center for Global Health and Medicine</s1><s2>Tokyo</s2><s3>JPN</s3><sZ>7 aut.</sZ></fA14><fA20><s1>11745-11753</s1></fA20><fA21><s1>2012</s1></fA21><fA23 i1="01"><s0>ENG</s0></fA23><fA43 i1="01"><s1>INIST</s1><s2>13592</s2><s5>354000506814370320</s5></fA43><fA44><s0>0000</s0><s1>© 2012 INIST-CNRS. All rights reserved.</s1></fA44><fA45><s0>49 ref.</s0></fA45><fA47 i1="01" i2="1"><s0>12-0410005</s0></fA47><fA60><s1>P</s1></fA60><fA61><s0>A</s0></fA61><fA64 i1="01" i2="1"><s0>Journal of virology</s0></fA64><fA66 i1="01"><s0>USA</s0></fA66><fC01 i1="01" l="ENG"><s0>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.</s0></fC01><fC02 i1="01" i2="X"><s0>002A05C10</s0></fC02><fC03 i1="01" i2="X" l="FRE"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="ENG"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="01" i2="X" l="SPA"><s0>Coronavirus</s0><s2>NW</s2><s5>01</s5></fC03><fC03 i1="02" i2="X" l="FRE"><s0>Protéine</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="ENG"><s0>Protein</s0><s5>05</s5></fC03><fC03 i1="02" i2="X" l="SPA"><s0>Proteína</s0><s5>05</s5></fC03><fC03 i1="03" i2="X" l="FRE"><s0>Réticulum endoplasmique</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="ENG"><s0>Endoplasmic reticulum</s0><s5>06</s5></fC03><fC03 i1="03" i2="X" l="SPA"><s0>Retículo endoplásmico</s0><s5>06</s5></fC03><fC03 i1="04" i2="X" l="FRE"><s0>Pouvoir infectant</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="ENG"><s0>Infectivity</s0><s5>07</s5></fC03><fC03 i1="04" i2="X" l="SPA"><s0>Poder infectante</s0><s5>07</s5></fC03><fC03 i1="05" i2="X" l="FRE"><s0>Syndrome respiratoire aigu sévère</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="05" i2="X" l="ENG"><s0>Severe acute respiratory syndrome</s0><s2>NM</s2><s5>14</s5></fC03><fC03 i1="05" i2="X" l="SPA"><s0>Síndrome respiratorio agudo severo</s0><s2>NM</s2><s5>14</s5></fC03><fC07 i1="01" i2="X" l="FRE"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="ENG"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="01" i2="X" l="SPA"><s0>Coronaviridae</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="FRE"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="ENG"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="02" i2="X" l="SPA"><s0>Nidovirales</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="FRE"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="ENG"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="03" i2="X" l="SPA"><s0>Virus</s0><s2>NW</s2></fC07><fC07 i1="04" i2="X" l="FRE"><s0>Pathologie de l'appareil respiratoire</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="ENG"><s0>Respiratory disease</s0><s5>13</s5></fC07><fC07 i1="04" i2="X" l="SPA"><s0>Aparato respiratorio patología</s0><s5>13</s5></fC07><fC07 i1="05" i2="X" l="FRE"><s0>Virose</s0></fC07><fC07 i1="05" i2="X" l="ENG"><s0>Viral disease</s0></fC07><fC07 i1="05" i2="X" l="SPA"><s0>Virosis</s0></fC07><fC07 i1="06" i2="X" l="FRE"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="ENG"><s0>Infection</s0></fC07><fC07 i1="06" i2="X" l="SPA"><s0>Infección</s0></fC07><fC07 i1="07" i2="X" l="FRE"><s0>Pathologie des poumons</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="ENG"><s0>Lung disease</s0><s5>16</s5></fC07><fC07 i1="07" i2="X" l="SPA"><s0>Pulmón patología</s0><s5>16</s5></fC07><fN21><s1>317</s1></fN21><fN44 i1="01"><s1>OTO</s1></fN44><fN82><s1>OTO</s1></fN82></pA></standard></inist><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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|texte= Monitoring of S Protein Maturation in the Endoplasmic Reticulum by Calnexin Is Important for the Infectivity of Severe Acute Respiratory Syndrome Coronavirus
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