Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus
Identifieur interne : 001E42 ( Ncbi/Merge ); précédent : 001E41; suivant : 001E43Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus
Auteurs : Wei-Chen Hsin [Taïwan] ; Chan-Hua Chang [Taïwan] ; Chi-You Chang [Taïwan] ; Wei-Hao Peng [Taïwan] ; Chung-Liang Chien [Taïwan] ; Ming-Fu Chang [Taïwan] ; Shin C. Chang [Taïwan]Source :
- Journal of Biomedical Science [ 1021-7770 ] ; 2018.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Assemblage viral.
- métabolisme : ARN messager, ARN viral, Protéines nucléocapside.
- physiologie : Coronavirus du syndrome respiratoire du Moyen-Orient.
- Cellules HEK293, Humains, Lignée cellulaire tumorale.
English descriptors
- KwdEn :
- MESH :
- chemical , metabolism : Nucleocapsid Proteins, RNA, Messenger, RNA, Viral.
- genetics : Virus Assembly.
- physiology : Middle East Respiratory Syndrome Coronavirus.
- Cell Line, Tumor, HEK293 Cells, Humans.
Abstract
Middle East respiratory syndrome coronavirus (MERS-CoV) consists of a positive-sense, single-stranded RNA genome and four structural proteins: the spike, envelope, membrane, and nucleocapsid protein. The assembly of the viral genome into virus particles involves viral structural proteins and is believed to be mediated through recognition of specific sequences and RNA structures of the viral genome.
A culture system for the production of MERS coronavirus-like particles (MERS VLPs) was determined and established by electron microscopy and the detection of coexpressed viral structural proteins. Using the VLP system, a 258-nucleotide RNA fragment, which spans nucleotides 19,712 to 19,969 of the MERS-CoV genome (designated PS258(19712–19969)ME), was identified to function as a packaging signal. Assembly of the RNA packaging signal into MERS VLPs is dependent on the viral nucleocapsid protein. In addition, a 45-nucleotide stable stem-loop substructure of the PS258(19712–19969)ME interacted with both the N-terminal domain and the C-terminal domain of the viral nucleocapsid protein. Furthermore, a functional SARS-CoV RNA packaging signal failed to assemble into the MERS VLPs, which indicated virus-specific assembly of the RNA genome.
A MERS-oV RNA packaging signal was identified by the detection of GFP expression following an incubation of MERS VLPs carrying the heterologous mRNA GFP-PS258(19712–19969)ME with virus permissive Huh7 cells. The MERS VLP system could help us in understanding virus infection and morphogenesis.
Url:
DOI: 10.1186/s12929-018-0449-x
PubMed: 29793506
PubMed Central: 5966903
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PMC:5966903Le document en format XML
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<term>Middle East Respiratory Syndrome Coronavirus (physiology)</term>
<term>Nucleocapsid Proteins (metabolism)</term>
<term>RNA, Messenger (metabolism)</term>
<term>RNA, Viral (metabolism)</term>
<term>Virus Assembly (genetics)</term>
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<term>ARN viral (métabolisme)</term>
<term>Assemblage viral (génétique)</term>
<term>Cellules HEK293</term>
<term>Coronavirus du syndrome respiratoire du Moyen-Orient (physiologie)</term>
<term>Humains</term>
<term>Lignée cellulaire tumorale</term>
<term>Protéines nucléocapside (métabolisme)</term>
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<term>RNA, Viral</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Virus Assembly</term>
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<term>ARN viral</term>
<term>Protéines nucléocapside</term>
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p id="Par1">Middle East respiratory syndrome coronavirus (MERS-CoV) consists of a positive-sense, single-stranded RNA genome and four structural proteins: the spike, envelope, membrane, and nucleocapsid protein. The assembly of the viral genome into virus particles involves viral structural proteins and is believed to be mediated through recognition of specific sequences and RNA structures of the viral genome.</p>
</sec>
<sec><title>Methods and Results</title>
<p id="Par2">A culture system for the production of MERS coronavirus-like particles (MERS VLPs) was determined and established by electron microscopy and the detection of coexpressed viral structural proteins. Using the VLP system, a 258-nucleotide RNA fragment, which spans nucleotides 19,712 to 19,969 of the MERS-CoV genome (designated PS258(19712–19969)<sub>ME</sub>
), was identified to function as a packaging signal. Assembly of the RNA packaging signal into MERS VLPs is dependent on the viral nucleocapsid protein. In addition, a 45-nucleotide stable stem-loop substructure of the PS258(19712–19969)<sub>ME</sub>
interacted with both the N-terminal domain and the C-terminal domain of the viral nucleocapsid protein. Furthermore, a functional SARS-CoV RNA packaging signal failed to assemble into the MERS VLPs, which indicated virus-specific assembly of the RNA genome.</p>
</sec>
<sec><title>Conclusions</title>
<p id="Par3">A MERS-oV RNA packaging signal was identified by the detection of GFP expression following an incubation of MERS VLPs carrying the heterologous mRNA GFP-PS258(19712–19969)<sub>ME</sub>
with virus permissive Huh7 cells. The MERS VLP system could help us in understanding virus infection and morphogenesis.</p>
</sec>
</div>
</front>
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No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
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<author><name sortKey="Chang, Chi You" sort="Chang, Chi You" uniqKey="Chang C" first="Chi-You" last="Chang">Chi-You Chang</name>
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<institution>Institute of Anatomy and Cell Biology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
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No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
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No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
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<author><name sortKey="Chang, Ming Fu" sort="Chang, Ming Fu" uniqKey="Chang M" first="Ming-Fu" last="Chang">Ming-Fu Chang</name>
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<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Biochemistry and Molecular Biology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chang, Shin C" sort="Chang, Shin C" uniqKey="Chang S" first="Shin C." last="Chang">Shin C. Chang</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Microbiology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
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<idno type="pmid">29793506</idno>
<idno type="pmc">5966903</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5966903</idno>
<idno type="RBID">PMC:5966903</idno>
<idno type="doi">10.1186/s12929-018-0449-x</idno>
<date when="2018">2018</date>
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<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000511</idno>
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus</title>
<author><name sortKey="Hsin, Wei Chen" sort="Hsin, Wei Chen" uniqKey="Hsin W" first="Wei-Chen" last="Hsin">Wei-Chen Hsin</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Microbiology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chang, Chan Hua" sort="Chang, Chan Hua" uniqKey="Chang C" first="Chan-Hua" last="Chang">Chan-Hua Chang</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Microbiology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chang, Chi You" sort="Chang, Chi You" uniqKey="Chang C" first="Chi-You" last="Chang">Chi-You Chang</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Microbiology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Peng, Wei Hao" sort="Peng, Wei Hao" uniqKey="Peng W" first="Wei-Hao" last="Peng">Wei-Hao Peng</name>
<affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Anatomy and Cell Biology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chien, Chung Liang" sort="Chien, Chung Liang" uniqKey="Chien C" first="Chung-Liang" last="Chien">Chung-Liang Chien</name>
<affiliation wicri:level="1"><nlm:aff id="Aff2"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Anatomy and Cell Biology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chang, Ming Fu" sort="Chang, Ming Fu" uniqKey="Chang M" first="Ming-Fu" last="Chang">Ming-Fu Chang</name>
<affiliation wicri:level="1"><nlm:aff id="Aff3"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Biochemistry and Molecular Biology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Chang, Shin C" sort="Chang, Shin C" uniqKey="Chang S" first="Shin C." last="Chang">Shin C. Chang</name>
<affiliation wicri:level="1"><nlm:aff id="Aff1"><institution-wrap><institution-id institution-id-type="ISNI">0000 0004 0546 0241</institution-id>
<institution-id institution-id-type="GRID">grid.19188.39</institution-id>
<institution>Institute of Microbiology, College of Medicine,</institution>
<institution>National Taiwan University,</institution>
</institution-wrap>
No. 1, Jen-Ai Road, First Section, Taipei, 100 Taiwan</nlm:aff>
<country xml:lang="fr">Taïwan</country>
<wicri:regionArea>No. 1, Jen-Ai Road, First Section, Taipei</wicri:regionArea>
<wicri:noRegion>Taipei</wicri:noRegion>
</affiliation>
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</analytic>
<series><title level="j">Journal of Biomedical Science</title>
<idno type="ISSN">1021-7770</idno>
<idno type="eISSN">1423-0127</idno>
<imprint><date when="2018">2018</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p id="Par1">Middle East respiratory syndrome coronavirus (MERS-CoV) consists of a positive-sense, single-stranded RNA genome and four structural proteins: the spike, envelope, membrane, and nucleocapsid protein. The assembly of the viral genome into virus particles involves viral structural proteins and is believed to be mediated through recognition of specific sequences and RNA structures of the viral genome.</p>
</sec>
<sec><title>Methods and Results</title>
<p id="Par2">A culture system for the production of MERS coronavirus-like particles (MERS VLPs) was determined and established by electron microscopy and the detection of coexpressed viral structural proteins. Using the VLP system, a 258-nucleotide RNA fragment, which spans nucleotides 19,712 to 19,969 of the MERS-CoV genome (designated PS258(19712–19969)<sub>ME</sub>
), was identified to function as a packaging signal. Assembly of the RNA packaging signal into MERS VLPs is dependent on the viral nucleocapsid protein. In addition, a 45-nucleotide stable stem-loop substructure of the PS258(19712–19969)<sub>ME</sub>
interacted with both the N-terminal domain and the C-terminal domain of the viral nucleocapsid protein. Furthermore, a functional SARS-CoV RNA packaging signal failed to assemble into the MERS VLPs, which indicated virus-specific assembly of the RNA genome.</p>
</sec>
<sec><title>Conclusions</title>
<p id="Par3">A MERS-oV RNA packaging signal was identified by the detection of GFP expression following an incubation of MERS VLPs carrying the heterologous mRNA GFP-PS258(19712–19969)<sub>ME</sub>
with virus permissive Huh7 cells. The MERS VLP system could help us in understanding virus infection and morphogenesis.</p>
</sec>
</div>
</front>
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</author>
<author><name sortKey="Chang, Chan Hua" sort="Chang, Chan Hua" uniqKey="Chang C" first="Chan-Hua" last="Chang">Chan-Hua Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan.</nlm:affiliation>
<country xml:lang="fr">Taïwan</country>
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</affiliation>
</author>
<author><name sortKey="Chang, Chi You" sort="Chang, Chi You" uniqKey="Chang C" first="Chi-You" last="Chang">Chi-You Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan.</nlm:affiliation>
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</author>
<author><name sortKey="Peng, Wei Hao" sort="Peng, Wei Hao" uniqKey="Peng W" first="Wei-Hao" last="Peng">Wei-Hao Peng</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Anatomy and Cell Biology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan.</nlm:affiliation>
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<author><name sortKey="Chien, Chung Liang" sort="Chien, Chung Liang" uniqKey="Chien C" first="Chung-Liang" last="Chien">Chung-Liang Chien</name>
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<wicri:regionArea>Institute of Anatomy and Cell Biology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100</wicri:regionArea>
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</author>
<author><name sortKey="Chang, Ming Fu" sort="Chang, Ming Fu" uniqKey="Chang M" first="Ming-Fu" last="Chang">Ming-Fu Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan. mfchang@ntu.edu.tw.</nlm:affiliation>
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<wicri:regionArea>Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100</wicri:regionArea>
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</affiliation>
</author>
<author><name sortKey="Chang, Shin C" sort="Chang, Shin C" uniqKey="Chang S" first="Shin C" last="Chang">Shin C. Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan. scchang093@ntu.edu.tw.</nlm:affiliation>
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<sourceDesc><biblStruct><analytic><title xml:lang="en">Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus.</title>
<author><name sortKey="Hsin, Wei Chen" sort="Hsin, Wei Chen" uniqKey="Hsin W" first="Wei-Chen" last="Hsin">Wei-Chen Hsin</name>
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<country xml:lang="fr">Taïwan</country>
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</affiliation>
</author>
<author><name sortKey="Chang, Chan Hua" sort="Chang, Chan Hua" uniqKey="Chang C" first="Chan-Hua" last="Chang">Chan-Hua Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan.</nlm:affiliation>
<country xml:lang="fr">Taïwan</country>
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</author>
<author><name sortKey="Chang, Chi You" sort="Chang, Chi You" uniqKey="Chang C" first="Chi-You" last="Chang">Chi-You Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan.</nlm:affiliation>
<country xml:lang="fr">Taïwan</country>
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</author>
<author><name sortKey="Peng, Wei Hao" sort="Peng, Wei Hao" uniqKey="Peng W" first="Wei-Hao" last="Peng">Wei-Hao Peng</name>
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<author><name sortKey="Chien, Chung Liang" sort="Chien, Chung Liang" uniqKey="Chien C" first="Chung-Liang" last="Chien">Chung-Liang Chien</name>
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</author>
<author><name sortKey="Chang, Ming Fu" sort="Chang, Ming Fu" uniqKey="Chang M" first="Ming-Fu" last="Chang">Ming-Fu Chang</name>
<affiliation wicri:level="1"><nlm:affiliation>Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan. mfchang@ntu.edu.tw.</nlm:affiliation>
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<affiliation wicri:level="1"><nlm:affiliation>Institute of Microbiology, College of Medicine, National Taiwan University, No. 1, Jen-Ai Road, First Section, Taipei, 100, Taiwan. scchang093@ntu.edu.tw.</nlm:affiliation>
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<series><title level="j">Journal of biomedical science</title>
<idno type="eISSN">1423-0127</idno>
<imprint><date when="2018" type="published">2018</date>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell Line, Tumor</term>
<term>HEK293 Cells</term>
<term>Humans</term>
<term>Middle East Respiratory Syndrome Coronavirus (physiology)</term>
<term>Nucleocapsid Proteins (metabolism)</term>
<term>RNA, Messenger (metabolism)</term>
<term>RNA, Viral (metabolism)</term>
<term>Virus Assembly (genetics)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ARN messager (métabolisme)</term>
<term>ARN viral (métabolisme)</term>
<term>Assemblage viral (génétique)</term>
<term>Cellules HEK293</term>
<term>Coronavirus du syndrome respiratoire du Moyen-Orient (physiologie)</term>
<term>Humains</term>
<term>Lignée cellulaire tumorale</term>
<term>Protéines nucléocapside (métabolisme)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Nucleocapsid Proteins</term>
<term>RNA, Messenger</term>
<term>RNA, Viral</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Virus Assembly</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Assemblage viral</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>ARN messager</term>
<term>ARN viral</term>
<term>Protéines nucléocapside</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Coronavirus du syndrome respiratoire du Moyen-Orient</term>
</keywords>
<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Middle East Respiratory Syndrome Coronavirus</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Cell Line, Tumor</term>
<term>HEK293 Cells</term>
<term>Humans</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Cellules HEK293</term>
<term>Humains</term>
<term>Lignée cellulaire tumorale</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Middle East respiratory syndrome coronavirus (MERS-CoV) consists of a positive-sense, single-stranded RNA genome and four structural proteins: the spike, envelope, membrane, and nucleocapsid protein. The assembly of the viral genome into virus particles involves viral structural proteins and is believed to be mediated through recognition of specific sequences and RNA structures of the viral genome.</div>
</front>
</TEI>
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