Interactions among SARS-CoV accessory proteins revealed by bimolecular fluorescence complementation assay
Identifieur interne : 000817 ( Pmc/Curation ); précédent : 000816; suivant : 000818Interactions among SARS-CoV accessory proteins revealed by bimolecular fluorescence complementation assay
Auteurs : Jianqiang Kong [République populaire de Chine] ; Yanwei Shi [République populaire de Chine] ; Zhifang Wang [République populaire de Chine] ; Yiting Pan [République populaire de Chine]Source :
- Acta Pharmaceutica Sinica. B [ 2211-3835 ] ; 2015.
Abstract
The accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b, 9b and ORF14), predicted unknown proteins (PUPs) encoded by the genes, are considered to be unique to the severe acute respiratory syndrome coronavirus (SARS-CoV) genome. These proteins play important roles in various biological processes mediated by interactions with their partners. However, very little is known about the interactions among these accessory proteins. Here, a EYFP (enhanced yellow fluorescent protein) bimolecular fluorescence complementation (BiFC) assay was used to detect the interactions among accessory proteins. 33 out of 81 interactions were identified by BiFC, much more than that identified by the yeast two-hybrid (Y2H) system. This is the first report describing direct visualization of interactions among accessory proteins of SARS-CoV. These findings attest to the general applicability of the BiFC system for the verification of protein-protein interactions.
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DOI: 10.1016/j.apsb.2015.05.002
PubMed: 26579480
PubMed Central: 4629423
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100050</wicri:regionArea></affiliation></author><author><name sortKey="Shi, Yanwei" sort="Shi, Yanwei" uniqKey="Shi Y" first="Yanwei" last="Shi">Yanwei Shi</name><affiliation wicri:level="1"><nlm:aff id="aff0005">State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Zhifang" sort="Wang, Zhifang" uniqKey="Wang Z" first="Zhifang" last="Wang">Zhifang Wang</name><affiliation wicri:level="1"><nlm:aff id="aff0005">State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050</wicri:regionArea></affiliation></author><author><name sortKey="Pan, Yiting" sort="Pan, Yiting" uniqKey="Pan Y" first="Yiting" last="Pan">Yiting Pan</name><affiliation wicri:level="1"><nlm:aff id="aff0010">School of Chemical Engineering, Beijing Institute of Petrochemical Technology, Beijing 102617, 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fluorescence complementation assay</title><author><name sortKey="Kong, Jianqiang" sort="Kong, Jianqiang" uniqKey="Kong J" first="Jianqiang" last="Kong">Jianqiang Kong</name><affiliation wicri:level="1"><nlm:aff id="aff0005">State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050</wicri:regionArea></affiliation></author><author><name sortKey="Shi, Yanwei" sort="Shi, Yanwei" uniqKey="Shi Y" first="Yanwei" last="Shi">Yanwei Shi</name><affiliation wicri:level="1"><nlm:aff id="aff0005">State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Zhifang" sort="Wang, Zhifang" uniqKey="Wang Z" first="Zhifang" last="Wang">Zhifang Wang</name><affiliation wicri:level="1"><nlm:aff id="aff0005">State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050</wicri:regionArea></affiliation></author><author><name sortKey="Pan, Yiting" sort="Pan, Yiting" uniqKey="Pan Y" first="Yiting" last="Pan">Yiting Pan</name><affiliation wicri:level="1"><nlm:aff id="aff0010">School of Chemical Engineering, Beijing Institute of Petrochemical Technology, Beijing 102617, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>School of Chemical Engineering, Beijing Institute of Petrochemical Technology, Beijing 102617</wicri:regionArea></affiliation></author></analytic><series><title level="j">Acta Pharmaceutica Sinica. B</title><idno type="ISSN">2211-3835</idno><idno type="eISSN">2211-3843</idno><imprint><date when="2015">2015</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b, 9b and ORF14), predicted unknown proteins (PUPs) encoded by the genes, are considered to be unique to the severe acute respiratory syndrome coronavirus (SARS-CoV) genome. These proteins play important roles in various biological processes mediated by interactions with their partners. However, very little is known about the interactions among these accessory proteins. Here, a EYFP (enhanced yellow fluorescent protein) bimolecular fluorescence complementation (BiFC) assay was used to detect the interactions among accessory proteins. 33 out of 81 interactions were identified by BiFC, much more than that identified by the yeast two-hybrid (Y2H) system. This is the first report describing direct visualization of interactions among accessory proteins of SARS-CoV. These findings attest to the general applicability of the BiFC system for the verification of protein-protein interactions.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Rota, Pa" uniqKey="Rota P">PA Rota</name></author><author><name sortKey="Oberste, Ms" uniqKey="Oberste M">MS Oberste</name></author><author><name sortKey="Monroe, Ss" uniqKey="Monroe S">SS Monroe</name></author><author><name sortKey="Nix, Wa" uniqKey="Nix W">WA Nix</name></author><author><name sortKey="Campagnoli, R" uniqKey="Campagnoli R">R Campagnoli</name></author><author><name sortKey="Icenogle, Jp" uniqKey="Icenogle J">JP Icenogle</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Marra, Ma" uniqKey="Marra M">MA Marra</name></author><author><name sortKey="Jones, Sj" uniqKey="Jones S">SJ Jones</name></author><author><name sortKey="Astell, Cr" uniqKey="Astell C">CR Astell</name></author><author><name sortKey="Holt, Ra" uniqKey="Holt R">RA Holt</name></author><author><name sortKey="Brooks Wilson, A" uniqKey="Brooks Wilson A">A Brooks-Wilson</name></author><author><name sortKey="Butterfield, Ys" uniqKey="Butterfield Y">YS Butterfield</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Tan, Yj" uniqKey="Tan Y">YJ Tan</name></author><author><name sortKey="Teng, E" uniqKey="Teng E">E Teng</name></author><author><name sortKey="Shen, S" uniqKey="Shen S">S Shen</name></author><author><name sortKey="Tan, Th" uniqKey="Tan T">TH Tan</name></author><author><name sortKey="Goh, Py" uniqKey="Goh P">PY Goh</name></author><author><name sortKey="Fielding, Bc" uniqKey="Fielding B">BC Fielding</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Calvo, E" uniqKey="Calvo E">E Calvo</name></author><author><name sortKey="Dediego, Ml" uniqKey="Dediego M">ML DeDiego</name></author><author><name sortKey="Garcia, P" uniqKey="Garcia P">P García</name></author><author><name sortKey="L Pez, Ja" uniqKey="L Pez J">JA López</name></author><author><name sortKey="Perez Bre A, P" uniqKey="Perez Bre A P">P Pérez-Breña</name></author><author><name sortKey="Falc N, A" uniqKey="Falc N A">A. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Acta Pharm Sin B</journal-id><journal-id journal-id-type="iso-abbrev">Acta Pharm Sin B</journal-id><journal-title-group><journal-title>Acta Pharmaceutica Sinica. B</journal-title></journal-title-group><issn pub-type="ppub">2211-3835</issn><issn pub-type="epub">2211-3843</issn><publisher><publisher-name>Elsevier</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">26579480</article-id><article-id pub-id-type="pmc">4629423</article-id><article-id pub-id-type="publisher-id">S2211-3835(15)00075-1</article-id><article-id pub-id-type="doi">10.1016/j.apsb.2015.05.002</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Interactions among SARS-CoV accessory proteins revealed by bimolecular fluorescence complementation assay</article-title></title-group><contrib-group><contrib contrib-type="author" id="au0005"><name><surname>Kong</surname><given-names>Jianqiang</given-names></name><email>jianqiangk@imm.ac.cn</email><xref rid="aff0005" ref-type="aff">a</xref><xref rid="cor1" ref-type="corresp">⁎</xref></contrib><contrib contrib-type="author" id="au0010"><name><surname>Shi</surname><given-names>Yanwei</given-names></name><xref rid="aff0005" ref-type="aff">a</xref></contrib><contrib contrib-type="author" id="au0015"><name><surname>Wang</surname><given-names>Zhifang</given-names></name><xref rid="aff0005" ref-type="aff">a</xref></contrib><contrib contrib-type="author" id="au0020"><name><surname>Pan</surname><given-names>Yiting</given-names></name><xref rid="aff0010" ref-type="aff">b</xref></contrib></contrib-group><aff id="aff0005"><label>a</label>State Key Laboratory of Bioactive Substance and Function of Natural Medicines & Ministry of Health Key Laboratory of Biosynthesis of Natural Products, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China</aff><aff id="aff0010"><label>b</label>School of Chemical Engineering, Beijing Institute of Petrochemical Technology, Beijing 102617, China</aff><author-notes><corresp id="cor1"><label>⁎</label>Corresponding author. Tel.: +86 10 63165169. <email>jianqiangk@imm.ac.cn</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>6</day><month>6</month><year>2015</year></pub-date><pmc-comment> PMC Release delay is 0 months and 0 days and was based on .</pmc-comment>
<pub-date pub-type="ppub"><month>9</month><year>2015</year></pub-date><pub-date pub-type="epub"><day>6</day><month>6</month><year>2015</year></pub-date><volume>5</volume><issue>5</issue><fpage>487</fpage><lpage>492</lpage><history><date date-type="received"><day>15</day><month>12</month><year>2014</year></date><date date-type="rev-recd"><day>13</day><month>4</month><year>2015</year></date><date date-type="accepted"><day>28</day><month>4</month><year>2015</year></date></history><permissions><copyright-statement>© 2015 Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences. Production and hosting by Elsevier B.V.</copyright-statement><copyright-year>2015</copyright-year><copyright-holder>Chinese Pharmaceutical Association and Institute of Materia Medica, Chinese Academy of Medical Sciences</copyright-holder><license><license-p>Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.</license-p></license></permissions><abstract id="ab0005"><p>The accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b, 9b and ORF14), predicted unknown proteins (PUPs) encoded by the genes, are considered to be unique to the severe acute respiratory syndrome coronavirus (SARS-CoV) genome. These proteins play important roles in various biological processes mediated by interactions with their partners. However, very little is known about the interactions among these accessory proteins. Here, a EYFP (enhanced yellow fluorescent protein) bimolecular fluorescence complementation (BiFC) assay was used to detect the interactions among accessory proteins. 33 out of 81 interactions were identified by BiFC, much more than that identified by the yeast two-hybrid (Y2H) system. This is the first report describing direct visualization of interactions among accessory proteins of SARS-CoV. These findings attest to the general applicability of the BiFC system for the verification of protein-protein interactions.</p></abstract><abstract abstract-type="graphical" id="ab0010"><title>Graphical abstract</title><p>Two non-fluorescent fragments (VN and VC) derived from a fluorescent protein EYFP were separately attached to two accessory proteins of SARS-CoV (APs) and the resulting fusion proteins were co-expressed in <italic>Saccharomyces cerevisiae</italic>. When the two accessory proteins interact, the split EYFP fragments can be functionally reconstituted, allowing a fluorescent readout. Thereby, the BiFC method can be used for direct visualization of accessory protein interactions in living cells.<fig id="f0010" position="anchor"><graphic xlink:href="fx1"></graphic></fig></p></abstract><kwd-group id="keys0005"><title>Abbreviations</title><kwd>aa, amino acids</kwd><kwd>AD, activation domain</kwd><kwd>BD, binding domain</kwd><kwd>BiFC, bimolecular fluorescence complementation</kwd><kwd>Co-IP, co-immunoprecipitation</kwd><kwd>E, envelope</kwd><kwd>EYFP, enhanced yellow fluorescent protein</kwd><kwd>M, membrane</kwd><kwd>N, nucleocapsid</kwd><kwd>NLS, nuclear localization signal</kwd><kwd>ORFs, open reading frames</kwd><kwd>PCR, polymerase chain reaction</kwd><kwd>PPIs, protein-protein interactions</kwd><kwd>PUPs, predicted unknown proteins</kwd><kwd>S, spike</kwd><kwd>SARS-CoV, severe acute respiratory syndrome coronavirus</kwd><kwd>Y2H, yeast two-hybrid</kwd></kwd-group><kwd-group id="keys0010"><title>KEY WORDS</title><kwd>SARS-CoV</kwd><kwd>Accessory proteins</kwd><kwd>Y2H</kwd><kwd>Bimolecular fluorescence complementation assay</kwd></kwd-group></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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