Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: Implications for assembly and vaccine production
Identifieur interne : 001839 ( Main/Exploration ); précédent : 001838; suivant : 001840Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: Implications for assembly and vaccine production
Auteurs : YUE HUANG [États-Unis] ; Zhi-Yong Yang [États-Unis] ; Wing-Pui Kong [États-Unis] ; Gary J. Nabel [États-Unis]Source :
- Journal of virology [ 0022-538X ] ; 2004.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Vaccin.
English descriptors
- KwdEn :
Abstract
The recently emerged severe acute respiratory syndrome coronavirus (SARS-CoV) contains four structural genes, two replicase-transcriptase open reading frames, and more than five potential genes of unknown function. Despite this relative simplicity, the molecular regulation of SARS-CoV replication and assembly is not understood. Here, we report that two viral genes, encoding the SARS-CoV membrane (M) and nucleocapsid (N) proteins, are necessary and sufficient for formation of virus-like particles. Expression vectors encoding these two proteins were synthesized by using preferred human codons. When M and N expression plasmids were cotransfected into human 293 renal epithelial cells, pseudoparticles formed readily. The addition of a third gene, encoding the spike (S) glycoprotein, facilitated budding of particles that contained a corona-like halo resembling SARS-CoV when examined by transmission electron microscopy, with a buoyant density characteristic of coronaviruses. Specific biochemical interactions of these proteins were also shown in vitro. The S, M, and N proteins of the SARS-CoV are, therefore, necessary and sufficient for pseudovirus assembly. These findings advance the understanding of the morphogenesis of SARS-CoV and enable the generation of safe, conformational mimetics of the SARS virus that may facilitate the development of vaccines and antiviral drugs.
Affiliations:
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Nabel</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Maryland</region></placeName></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">INIST</idno><idno type="inist">05-0000571</idno><date when="2004">2004</date><idno type="stanalyst">PASCAL 05-0000571 INIST</idno><idno type="RBID">Pascal:05-0000571</idno><idno type="wicri:Area/PascalFrancis/Corpus">000052</idno><idno type="wicri:Area/PascalFrancis/Curation">000007</idno><idno type="wicri:Area/PascalFrancis/Checkpoint">000052</idno><idno type="wicri:explorRef" wicri:stream="PascalFrancis" wicri:step="Checkpoint">000052</idno><idno type="wicri:doubleKey">0022-538X:2004:Yue Huang:generation:of:synthetic</idno><idno type="wicri:Area/Main/Merge">001850</idno><idno type="wicri:Area/Main/Curation">001839</idno><idno type="wicri:Area/Main/Exploration">001839</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Generation of synthetic severe acute respiratory syndrome coronavirus pseudoparticles: Implications for assembly and vaccine production</title><author><name sortKey="Yue Huang" sort="Yue Huang" uniqKey="Yue Huang" last="Yue Huang">YUE HUANG</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Maryland</region></placeName></affiliation></author><author><name sortKey="Yang, Zhi Yong" sort="Yang, Zhi Yong" uniqKey="Yang Z" first="Zhi-Yong" last="Yang">Zhi-Yong Yang</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Maryland</region></placeName></affiliation></author><author><name sortKey="Kong, Wing Pui" sort="Kong, Wing Pui" uniqKey="Kong W" first="Wing-Pui" last="Kong">Wing-Pui Kong</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Maryland</region></placeName></affiliation></author><author><name sortKey="Nabel, Gary J" sort="Nabel, Gary J" uniqKey="Nabel G" first="Gary J." last="Nabel">Gary J. Nabel</name><affiliation wicri:level="2"><inist:fA14 i1="01"><s1>Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health</s1><s2>Bethesda, Maryland</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><placeName><region type="state">Maryland</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="2004">2004</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>Acute</term><term>Coronavirus</term><term>Critically ill</term><term>Immunoprophylaxis</term><term>Microbiology</term><term>Prevention</term><term>Respiratory tract</term><term>Severe</term><term>Severe acute respiratory syndrome</term><term>Vaccine</term><term>Virology</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term><term>Grave</term><term>Malade état grave</term><term>Aigu</term><term>Voie respiratoire</term><term>Vaccin</term><term>Prévention</term><term>Immunoprophylaxie</term><term>Microbiologie</term><term>Syndrome respiratoire aigu sévère</term><term>Virologie</term><term>Forme grave</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Vaccin</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The recently emerged severe acute respiratory syndrome coronavirus (SARS-CoV) contains four structural genes, two replicase-transcriptase open reading frames, and more than five potential genes of unknown function. Despite this relative simplicity, the molecular regulation of SARS-CoV replication and assembly is not understood. Here, we report that two viral genes, encoding the SARS-CoV membrane (M) and nucleocapsid (N) proteins, are necessary and sufficient for formation of virus-like particles. Expression vectors encoding these two proteins were synthesized by using preferred human codons. When M and N expression plasmids were cotransfected into human 293 renal epithelial cells, pseudoparticles formed readily. The addition of a third gene, encoding the spike (S) glycoprotein, facilitated budding of particles that contained a corona-like halo resembling SARS-CoV when examined by transmission electron microscopy, with a buoyant density characteristic of coronaviruses. Specific biochemical interactions of these proteins were also shown in vitro. The S, M, and N proteins of the SARS-CoV are, therefore, necessary and sufficient for pseudovirus assembly. These findings advance the understanding of the morphogenesis of SARS-CoV and enable the generation of safe, conformational mimetics of the SARS virus that may facilitate the development of vaccines and antiviral drugs.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Maryland</li></region></list><tree><country name="États-Unis"><region name="Maryland"><name sortKey="Yue Huang" sort="Yue Huang" uniqKey="Yue Huang" last="Yue Huang">YUE HUANG</name></region><name sortKey="Kong, Wing Pui" sort="Kong, Wing Pui" uniqKey="Kong W" first="Wing-Pui" last="Kong">Wing-Pui Kong</name><name sortKey="Nabel, Gary J" sort="Nabel, Gary J" uniqKey="Nabel G" first="Gary J." last="Nabel">Gary J. Nabel</name><name sortKey="Yang, Zhi Yong" sort="Yang, Zhi Yong" uniqKey="Yang Z" first="Zhi-Yong" last="Yang">Zhi-Yong Yang</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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