Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus
Identifieur interne : 000511 ( Pmc/Checkpoint ); précédent : 000510; suivant : 000512Nucleocapsid 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.
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
Affiliations:
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PMC:5966903Le document en format XML
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<series><title level="j">Journal of Biomedical Science</title>
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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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<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Biomed Sci</journal-id>
<journal-id journal-id-type="iso-abbrev">J. Biomed. Sci</journal-id>
<journal-title-group><journal-title>Journal of Biomedical Science</journal-title>
</journal-title-group>
<issn pub-type="ppub">1021-7770</issn>
<issn pub-type="epub">1423-0127</issn>
<publisher><publisher-name>BioMed Central</publisher-name>
<publisher-loc>London</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">29793506</article-id>
<article-id pub-id-type="pmc">5966903</article-id>
<article-id pub-id-type="publisher-id">449</article-id>
<article-id pub-id-type="doi">10.1186/s12929-018-0449-x</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Research</subject>
</subj-group>
</article-categories>
<title-group><article-title>Nucleocapsid protein-dependent assembly of the RNA packaging signal of Middle East respiratory syndrome coronavirus</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Hsin</surname>
<given-names>Wei-Chen</given-names>
</name>
<address><email>rabbit6350903@gmail.com</email>
</address>
<xref ref-type="aff" rid="Aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chang</surname>
<given-names>Chan-Hua</given-names>
</name>
<address><email>dikefither@gmail.com</email>
</address>
<xref ref-type="aff" rid="Aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chang</surname>
<given-names>Chi-You</given-names>
</name>
<address><email>r02445124@ntu.edu.tw</email>
</address>
<xref ref-type="aff" rid="Aff1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Peng</surname>
<given-names>Wei-Hao</given-names>
</name>
<address><email>pengweihao@ntu.edu.tw</email>
</address>
<xref ref-type="aff" rid="Aff2">2</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chien</surname>
<given-names>Chung-Liang</given-names>
</name>
<address><email>Chien@ntu.edu.tw</email>
</address>
<xref ref-type="aff" rid="Aff2">2</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><name><surname>Chang</surname>
<given-names>Ming-Fu</given-names>
</name>
<address><phone>886-2-23123456 ext. 88217</phone>
<email>mfchang@ntu.edu.tw</email>
</address>
<xref ref-type="aff" rid="Aff3">3</xref>
</contrib>
<contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-0365-8364</contrib-id>
<name><surname>Chang</surname>
<given-names>Shin C.</given-names>
</name>
<address><phone>886-2-23123456 ext. 88290</phone>
<email>scchang093@ntu.edu.tw</email>
</address>
<xref ref-type="aff" rid="Aff1">1</xref>
</contrib>
<aff id="Aff1"><label>1</label>
<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</aff>
<aff id="Aff2"><label>2</label>
<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</aff>
<aff id="Aff3"><label>3</label>
<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</aff>
</contrib-group>
<pub-date pub-type="epub"><day>24</day>
<month>5</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="pmc-release"><day>24</day>
<month>5</month>
<year>2018</year>
</pub-date>
<pub-date pub-type="collection"><year>2018</year>
</pub-date>
<volume>25</volume>
<elocation-id>47</elocation-id>
<history><date date-type="received"><day>1</day>
<month>12</month>
<year>2017</year>
</date>
<date date-type="accepted"><day>15</day>
<month>5</month>
<year>2018</year>
</date>
</history>
<permissions><copyright-statement>© The Author(s). 2018</copyright-statement>
<license license-type="OpenAccess"><license-p><bold>Open Access</bold>
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>
), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/publicdomain/zero/1.0/">http://creativecommons.org/publicdomain/zero/1.0/</ext-link>
) applies to the data made available in this article, unless otherwise stated.</license-p>
</license>
</permissions>
<abstract id="Abs1"><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>
</abstract>
<kwd-group xml:lang="en"><title>Keywords</title>
<kwd>MERS-CoV</kwd>
<kwd>RNA packaging signal</kwd>
<kwd>Nucleocapsid protein</kwd>
</kwd-group>
<funding-group><award-group><funding-source><institution-wrap><institution-id institution-id-type="FundRef">http://dx.doi.org/10.13039/501100004663</institution-id>
<institution>Ministry of Science and Technology, Taiwan</institution>
</institution-wrap>
</funding-source>
<award-id>102-2321-B-002-028</award-id>
<award-id>107-2321-B-002-023</award-id>
<principal-award-recipient><name><surname>Chang</surname>
<given-names>Ming-Fu</given-names>
</name>
</principal-award-recipient>
</award-group>
</funding-group>
<custom-meta-group><custom-meta><meta-name>issue-copyright-statement</meta-name>
<meta-value>© The Author(s) 2018</meta-value>
</custom-meta>
</custom-meta-group>
</article-meta>
</front>
</pmc>
<affiliations><list><country><li>Taïwan</li>
</country>
</list>
<tree><country name="Taïwan"><noRegion><name sortKey="Hsin, Wei Chen" sort="Hsin, Wei Chen" uniqKey="Hsin W" first="Wei-Chen" last="Hsin">Wei-Chen Hsin</name>
</noRegion>
<name sortKey="Chang, Chan Hua" sort="Chang, Chan Hua" uniqKey="Chang C" first="Chan-Hua" last="Chang">Chan-Hua Chang</name>
<name sortKey="Chang, Chi You" sort="Chang, Chi You" uniqKey="Chang C" first="Chi-You" last="Chang">Chi-You Chang</name>
<name sortKey="Chang, Ming Fu" sort="Chang, Ming Fu" uniqKey="Chang M" first="Ming-Fu" last="Chang">Ming-Fu Chang</name>
<name sortKey="Chang, Shin C" sort="Chang, Shin C" uniqKey="Chang S" first="Shin C." last="Chang">Shin C. Chang</name>
<name sortKey="Chien, Chung Liang" sort="Chien, Chung Liang" uniqKey="Chien C" first="Chung-Liang" last="Chien">Chung-Liang Chien</name>
<name sortKey="Peng, Wei Hao" sort="Peng, Wei Hao" uniqKey="Peng W" first="Wei-Hao" last="Peng">Wei-Hao Peng</name>
</country>
</tree>
</affiliations>
</record>
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