Expression cloning of functional receptor used by SARS coronavirus.
Identifieur interne : 005356 ( Main/Exploration ); précédent : 005355; suivant : 005357Expression cloning of functional receptor used by SARS coronavirus.
Auteurs : Peigang Wang [République populaire de Chine] ; Jian Chen ; Aihua Zheng ; Yuchun Nie ; Xuanling Shi ; Wei Wang ; Guangwen Wang ; Min Luo ; Huijun Liu ; Lei Tan ; Xijun Song ; Zai Wang ; Xiaolei Yin ; Xiuxia Qu ; Xiaojing Wang ; Tingting Qing ; Mingxiao Ding ; Hongkui DengSource :
- Biochemical and biophysical research communications [ 0006-291X ] ; 2004.
Descripteurs français
- KwdFr :
- ADN complémentaire (métabolisme), Animaux, Banque de gènes, Carboxypeptidases (métabolisme), Cellules HeLa, Cellules NIH 3T3, Cellules Vero, Clonage moléculaire, Cytométrie en flux, Glycoprotéines (), Glycoprotéines membranaires (métabolisme), Humains, Liaison aux protéines, Ligands, Modèles génétiques, Peptidyl-Dipeptidase A, Protéines de l'enveloppe virale, Protéines de transport, RT-PCR, Retroviridae (génétique), Réaction de polymérisation en chaîne, Récepteurs viraux (), Souris, Structure tertiaire des protéines, Séparation cellulaire, Séquences répétées terminales, Virus du SRAS (métabolisme).
- MESH :
- génétique : Retroviridae.
- métabolisme : ADN complémentaire, Carboxypeptidases, Glycoprotéines membranaires, Virus du SRAS.
- Animaux, Banque de gènes, Cellules HeLa, Cellules NIH 3T3, Cellules Vero, Clonage moléculaire, Cytométrie en flux, Glycoprotéines, Humains, Liaison aux protéines, Ligands, Modèles génétiques, Peptidyl-Dipeptidase A, Protéines de l'enveloppe virale, Protéines de transport, RT-PCR, Réaction de polymérisation en chaîne, Récepteurs viraux, Souris, Structure tertiaire des protéines, Séparation cellulaire, Séquences répétées terminales.
English descriptors
- KwdEn :
- Animals, Carboxypeptidases (metabolism), Carrier Proteins, Cell Separation, Chlorocebus aethiops, Cloning, Molecular, DNA, Complementary (metabolism), Flow Cytometry, Gene Library, Glycoproteins (chemistry), HeLa Cells, Humans, Ligands, Membrane Glycoproteins (metabolism), Mice, Models, Genetic, NIH 3T3 Cells, Peptidyl-Dipeptidase A, Polymerase Chain Reaction, Protein Binding, Protein Structure, Tertiary, Receptors, Virus (chemistry), Retroviridae (genetics), Reverse Transcriptase Polymerase Chain Reaction, SARS Virus (metabolism), Terminal Repeat Sequences, Vero Cells, Viral Envelope Proteins.
- MESH :
- chemical , chemistry : Glycoproteins, Receptors, Virus.
- chemical , metabolism : Carboxypeptidases, DNA, Complementary, Membrane Glycoproteins.
- genetics : Retroviridae.
- metabolism : SARS Virus.
- Animals, Carrier Proteins, Cell Separation, Chlorocebus aethiops, Cloning, Molecular, Flow Cytometry, Gene Library, HeLa Cells, Humans, Ligands, Mice, Models, Genetic, NIH 3T3 Cells, Peptidyl-Dipeptidase A, Polymerase Chain Reaction, Protein Binding, Protein Structure, Tertiary, Reverse Transcriptase Polymerase Chain Reaction, Terminal Repeat Sequences, Vero Cells, Viral Envelope Proteins.
Abstract
We have expressed a series of truncated spike (S) glycoproteins of SARS-CoV and found that the N-terminus 14-502 residuals were sufficient to bind to SARS-CoV susceptible Vero E6 cells. With this soluble S protein fragment as an affinity ligand, we screened HeLa cells transduced with retroviral cDNA library from Vero E6 cells and obtained a HeLa cell clone which could bind with the S protein. This cell clone was susceptible to HIV/SARS pseudovirus infection and the presence of a functional receptor for S protein in this cell clone was confirmed by the cell-cell fusion assay. Further studies showed the susceptibility of this cell was due to the expression of endogenous angiotensin-converting enzyme 2 (ACE2) which was activated by inserted LTR from retroviral vector used for expression cloning. When human ACE2 cDNA was transduced into NIH3T3 cells, the ACE2 expressing NIH3T3 cells could be infected with HIV/SARS pseudovirus. These data clearly demonstrated that ACE2 was the functional receptor for SARS-CoV.
DOI: 10.1016/j.bbrc.2004.01.076
PubMed: 14766227
Affiliations:
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Le document en format XML
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type="wicri:Area/Main/Curation">005356</idno><idno type="wicri:Area/Main/Exploration">005356</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Expression cloning of functional receptor used by SARS coronavirus.</title><author><name sortKey="Wang, Peigang" sort="Wang, Peigang" uniqKey="Wang P" first="Peigang" last="Wang">Peigang Wang</name><affiliation wicri:level="4"><nlm:affiliation>Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, PR China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871</wicri:regionArea><placeName><settlement type="city">Pékin</settlement></placeName><orgName type="university">Université de Pékin</orgName><placeName><settlement type="city">Pékin</settlement><region type="capitale">Pékin</region></placeName></affiliation></author><author><name sortKey="Chen, Jian" sort="Chen, Jian" uniqKey="Chen J" first="Jian" last="Chen">Jian Chen</name></author><author><name sortKey="Zheng, Aihua" sort="Zheng, Aihua" uniqKey="Zheng A" first="Aihua" last="Zheng">Aihua Zheng</name></author><author><name sortKey="Nie, Yuchun" sort="Nie, Yuchun" uniqKey="Nie Y" first="Yuchun" last="Nie">Yuchun Nie</name></author><author><name sortKey="Shi, Xuanling" sort="Shi, Xuanling" uniqKey="Shi X" first="Xuanling" last="Shi">Xuanling Shi</name></author><author><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name></author><author><name sortKey="Wang, Guangwen" sort="Wang, Guangwen" uniqKey="Wang G" first="Guangwen" last="Wang">Guangwen Wang</name></author><author><name sortKey="Luo, Min" sort="Luo, Min" uniqKey="Luo M" first="Min" last="Luo">Min Luo</name></author><author><name sortKey="Liu, Huijun" sort="Liu, Huijun" uniqKey="Liu H" first="Huijun" last="Liu">Huijun Liu</name></author><author><name sortKey="Tan, Lei" sort="Tan, Lei" uniqKey="Tan L" first="Lei" last="Tan">Lei Tan</name></author><author><name sortKey="Song, Xijun" sort="Song, Xijun" uniqKey="Song X" first="Xijun" last="Song">Xijun Song</name></author><author><name sortKey="Wang, Zai" sort="Wang, Zai" uniqKey="Wang Z" first="Zai" last="Wang">Zai Wang</name></author><author><name sortKey="Yin, Xiaolei" sort="Yin, Xiaolei" uniqKey="Yin X" first="Xiaolei" last="Yin">Xiaolei Yin</name></author><author><name sortKey="Qu, Xiuxia" sort="Qu, Xiuxia" uniqKey="Qu X" first="Xiuxia" last="Qu">Xiuxia Qu</name></author><author><name sortKey="Wang, Xiaojing" sort="Wang, Xiaojing" uniqKey="Wang X" first="Xiaojing" last="Wang">Xiaojing Wang</name></author><author><name sortKey="Qing, Tingting" sort="Qing, Tingting" uniqKey="Qing T" first="Tingting" last="Qing">Tingting Qing</name></author><author><name sortKey="Ding, Mingxiao" sort="Ding, Mingxiao" uniqKey="Ding M" first="Mingxiao" last="Ding">Mingxiao Ding</name></author><author><name sortKey="Deng, Hongkui" sort="Deng, Hongkui" uniqKey="Deng H" first="Hongkui" last="Deng">Hongkui Deng</name></author></analytic><series><title level="j">Biochemical and biophysical research communications</title><idno type="ISSN">0006-291X</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Carboxypeptidases (metabolism)</term><term>Carrier Proteins</term><term>Cell Separation</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>DNA, Complementary (metabolism)</term><term>Flow Cytometry</term><term>Gene Library</term><term>Glycoproteins (chemistry)</term><term>HeLa Cells</term><term>Humans</term><term>Ligands</term><term>Membrane Glycoproteins (metabolism)</term><term>Mice</term><term>Models, Genetic</term><term>NIH 3T3 Cells</term><term>Peptidyl-Dipeptidase A</term><term>Polymerase Chain Reaction</term><term>Protein Binding</term><term>Protein Structure, Tertiary</term><term>Receptors, Virus (chemistry)</term><term>Retroviridae (genetics)</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>SARS Virus (metabolism)</term><term>Terminal Repeat Sequences</term><term>Vero Cells</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ADN complémentaire (métabolisme)</term><term>Animaux</term><term>Banque de gènes</term><term>Carboxypeptidases (métabolisme)</term><term>Cellules HeLa</term><term>Cellules NIH 3T3</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Cytométrie en flux</term><term>Glycoprotéines ()</term><term>Glycoprotéines membranaires (métabolisme)</term><term>Humains</term><term>Liaison aux protéines</term><term>Ligands</term><term>Modèles génétiques</term><term>Peptidyl-Dipeptidase A</term><term>Protéines de l'enveloppe virale</term><term>Protéines de transport</term><term>RT-PCR</term><term>Retroviridae (génétique)</term><term>Réaction de polymérisation en chaîne</term><term>Récepteurs viraux ()</term><term>Souris</term><term>Structure tertiaire des protéines</term><term>Séparation cellulaire</term><term>Séquences répétées terminales</term><term>Virus du SRAS (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Glycoproteins</term><term>Receptors, Virus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Carboxypeptidases</term><term>DNA, Complementary</term><term>Membrane Glycoproteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Retroviridae</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Retroviridae</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ADN complémentaire</term><term>Carboxypeptidases</term><term>Glycoprotéines membranaires</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Carrier Proteins</term><term>Cell Separation</term><term>Chlorocebus aethiops</term><term>Cloning, Molecular</term><term>Flow Cytometry</term><term>Gene Library</term><term>HeLa Cells</term><term>Humans</term><term>Ligands</term><term>Mice</term><term>Models, Genetic</term><term>NIH 3T3 Cells</term><term>Peptidyl-Dipeptidase A</term><term>Polymerase Chain Reaction</term><term>Protein Binding</term><term>Protein Structure, Tertiary</term><term>Reverse Transcriptase Polymerase Chain Reaction</term><term>Terminal Repeat Sequences</term><term>Vero Cells</term><term>Viral Envelope Proteins</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Banque de gènes</term><term>Cellules HeLa</term><term>Cellules NIH 3T3</term><term>Cellules Vero</term><term>Clonage moléculaire</term><term>Cytométrie en flux</term><term>Glycoprotéines</term><term>Humains</term><term>Liaison aux protéines</term><term>Ligands</term><term>Modèles génétiques</term><term>Peptidyl-Dipeptidase A</term><term>Protéines de l'enveloppe virale</term><term>Protéines de transport</term><term>RT-PCR</term><term>Réaction de polymérisation en chaîne</term><term>Récepteurs viraux</term><term>Souris</term><term>Structure tertiaire des protéines</term><term>Séparation cellulaire</term><term>Séquences répétées terminales</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We have expressed a series of truncated spike (S) glycoproteins of SARS-CoV and found that the N-terminus 14-502 residuals were sufficient to bind to SARS-CoV susceptible Vero E6 cells. With this soluble S protein fragment as an affinity ligand, we screened HeLa cells transduced with retroviral cDNA library from Vero E6 cells and obtained a HeLa cell clone which could bind with the S protein. This cell clone was susceptible to HIV/SARS pseudovirus infection and the presence of a functional receptor for S protein in this cell clone was confirmed by the cell-cell fusion assay. Further studies showed the susceptibility of this cell was due to the expression of endogenous angiotensin-converting enzyme 2 (ACE2) which was activated by inserted LTR from retroviral vector used for expression cloning. When human ACE2 cDNA was transduced into NIH3T3 cells, the ACE2 expressing NIH3T3 cells could be infected with HIV/SARS pseudovirus. These data clearly demonstrated that ACE2 was the functional receptor for SARS-CoV.</div></front></TEI><affiliations><list><country><li>République populaire de Chine</li></country><region><li>Pékin</li></region><settlement><li>Pékin</li></settlement><orgName><li>Université de Pékin</li></orgName></list><tree><noCountry><name sortKey="Chen, Jian" sort="Chen, Jian" uniqKey="Chen J" first="Jian" last="Chen">Jian Chen</name><name sortKey="Deng, Hongkui" sort="Deng, Hongkui" uniqKey="Deng H" first="Hongkui" last="Deng">Hongkui Deng</name><name sortKey="Ding, Mingxiao" sort="Ding, Mingxiao" uniqKey="Ding M" first="Mingxiao" last="Ding">Mingxiao Ding</name><name sortKey="Liu, Huijun" sort="Liu, Huijun" uniqKey="Liu H" first="Huijun" last="Liu">Huijun Liu</name><name sortKey="Luo, Min" sort="Luo, Min" uniqKey="Luo M" first="Min" last="Luo">Min Luo</name><name sortKey="Nie, Yuchun" sort="Nie, Yuchun" uniqKey="Nie Y" first="Yuchun" last="Nie">Yuchun Nie</name><name sortKey="Qing, Tingting" sort="Qing, Tingting" uniqKey="Qing T" first="Tingting" last="Qing">Tingting Qing</name><name sortKey="Qu, Xiuxia" sort="Qu, Xiuxia" uniqKey="Qu X" first="Xiuxia" last="Qu">Xiuxia Qu</name><name sortKey="Shi, Xuanling" sort="Shi, Xuanling" uniqKey="Shi X" first="Xuanling" last="Shi">Xuanling Shi</name><name sortKey="Song, Xijun" sort="Song, Xijun" uniqKey="Song X" first="Xijun" last="Song">Xijun Song</name><name sortKey="Tan, Lei" sort="Tan, Lei" uniqKey="Tan L" first="Lei" last="Tan">Lei Tan</name><name sortKey="Wang, Guangwen" sort="Wang, Guangwen" uniqKey="Wang G" first="Guangwen" last="Wang">Guangwen Wang</name><name sortKey="Wang, Wei" sort="Wang, Wei" uniqKey="Wang W" first="Wei" last="Wang">Wei Wang</name><name sortKey="Wang, Xiaojing" sort="Wang, Xiaojing" uniqKey="Wang X" first="Xiaojing" last="Wang">Xiaojing Wang</name><name sortKey="Wang, Zai" sort="Wang, Zai" uniqKey="Wang Z" first="Zai" last="Wang">Zai Wang</name><name sortKey="Yin, Xiaolei" sort="Yin, Xiaolei" uniqKey="Yin X" first="Xiaolei" last="Yin">Xiaolei Yin</name><name sortKey="Zheng, Aihua" sort="Zheng, Aihua" uniqKey="Zheng A" first="Aihua" last="Zheng">Aihua Zheng</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Wang, Peigang" sort="Wang, Peigang" uniqKey="Wang P" first="Peigang" last="Wang">Peigang Wang</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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