Biochemical evidence for the presence of mixed membrane topologies of the severe acute respiratory syndrome coronavirus envelope protein expressed in mammalian cells
Identifieur interne : 000972 ( Pmc/Checkpoint ); précédent : 000971; suivant : 000973Biochemical evidence for the presence of mixed membrane topologies of the severe acute respiratory syndrome coronavirus envelope protein expressed in mammalian cells
Auteurs : Q. Yuan [Singapour] ; Y. Liao [Singapour] ; J. Torres [Singapour] ; J. P. Tam [Singapour] ; D. X. Liu [Singapour]Source :
- Febs Letters [ 0014-5793 ] ; 2006.
Abstract
Coronavirus envelope (E) protein is a small integral membrane protein with multi-functions in virion assembly, morphogenesis and virus–host interaction. Different coronavirus E proteins share striking similarities in biochemical properties and biological functions, but seem to adopt distinct membrane topology. In this report, we study the membrane topology of the SARS-CoV E protein by immunofluorescent staining of cells differentially permeabilized with detergents and proteinase K protection assay. It was revealed that both the N- and C-termini of the SARS-CoV E protein are exposed to the cytoplasmic side of the membranes (NcytoCcyto). In contrast, parallel experiments showed that the E protein from infectious bronchitis virus (IBV) spanned the membranes once, with the N-terminus exposed luminally and the C-terminus exposed cytoplasmically (Nexo(lum)Ccyto). Intriguingly, a minor proportion of the SARS-CoV E protein was found to be modified by N-linked glycosylation on Asn 66 and inserted into the membranes once with the C-terminus exposed to the luminal side. The presence of two distinct membrane topologies of the SARS-CoV E protein may provide a useful clue to the pathogenesis of SARS-CoV.
Url:
DOI: 10.1016/j.febslet.2006.04.076
PubMed: 16684538
PubMed Central: 7094218
Affiliations:
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Liao</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Torres, J" sort="Torres, J" uniqKey="Torres J" first="J." last="Torres">J. Torres</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Tam, J P" sort="Tam, J P" uniqKey="Tam J" first="J. P." last="Tam">J. P. Tam</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Liu, D X" sort="Liu, D X" uniqKey="Liu D" first="D. X." last="Liu">D. X. 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Yuan</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Liao, Y" sort="Liao, Y" uniqKey="Liao Y" first="Y." last="Liao">Y. Liao</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Torres, J" sort="Torres, J" uniqKey="Torres J" first="J." last="Torres">J. Torres</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Tam, J P" sort="Tam, J P" uniqKey="Tam J" first="J. P." last="Tam">J. P. Tam</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation></author><author><name sortKey="Liu, D X" sort="Liu, D X" uniqKey="Liu D" first="D. X." last="Liu">D. X. Liu</name><affiliation wicri:level="1"><nlm:aff id="aff1">School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551</wicri:regionArea><wicri:noRegion>Singapore 637551</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="aff2">Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673, Singapore</nlm:aff><country xml:lang="fr">Singapour</country><wicri:regionArea>Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673</wicri:regionArea><wicri:noRegion>Singapore 138673</wicri:noRegion></affiliation></author></analytic><series><title level="j">Febs Letters</title><idno type="ISSN">0014-5793</idno><idno type="eISSN">1873-3468</idno><imprint><date when="2006">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Coronavirus envelope (E) protein is a small integral membrane protein with multi-functions in virion assembly, morphogenesis and virus–host interaction. Different coronavirus E proteins share striking similarities in biochemical properties and biological functions, but seem to adopt distinct membrane topology. In this report, we study the membrane topology of the SARS-CoV E protein by immunofluorescent staining of cells differentially permeabilized with detergents and proteinase K protection assay. It was revealed that both the N- and C-termini of the SARS-CoV E protein are exposed to the cytoplasmic side of the membranes (N<sub>cyto</sub>C<sub>cyto</sub>). In contrast, parallel experiments showed that the E protein from infectious bronchitis virus (IBV) spanned the membranes once, with the N-terminus exposed luminally and the C-terminus exposed cytoplasmically (N<sub>exo(lum)</sub>C<sub>cyto</sub>). Intriguingly, a minor proportion of the SARS-CoV E protein was found to be modified by N-linked glycosylation on Asn 66 and inserted into the membranes once with the C-terminus exposed to the luminal side. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">FEBS Lett</journal-id><journal-id journal-id-type="iso-abbrev">FEBS Lett</journal-id><journal-title-group><journal-title>Febs Letters</journal-title></journal-title-group><issn pub-type="ppub">0014-5793</issn><issn pub-type="epub">1873-3468</issn><publisher><publisher-name>John Wiley & Sons Ltd</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">16684538</article-id><article-id pub-id-type="pmc">7094218</article-id><article-id pub-id-type="publisher-id">S0014-5793(06)00533-3</article-id><article-id pub-id-type="doi">10.1016/j.febslet.2006.04.076</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>Biochemical evidence for the presence of mixed membrane topologies of the severe acute respiratory syndrome coronavirus envelope protein expressed in mammalian cells</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Yuan</surname><given-names>Q.</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib><contrib contrib-type="author"><name><surname>Liao</surname><given-names>Y.</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib><contrib contrib-type="author"><name><surname>Torres</surname><given-names>J.</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib><contrib contrib-type="author"><name><surname>Tam</surname><given-names>J.P.</given-names></name><xref rid="aff1" ref-type="aff">a</xref></contrib><contrib contrib-type="author"><name><surname>Liu</surname><given-names>D.X.</given-names></name><email>dxliu@imcb.a-star.edu.sg</email><xref rid="aff1" ref-type="aff">a</xref><xref rid="aff2" ref-type="aff">b</xref><xref rid="cor1" ref-type="corresp">⁎</xref></contrib></contrib-group><aff id="aff1"><label>a</label>School of Biological Science, Nanyang Technological University, 60 Nanyang Drive, Singapore 637551, Singapore</aff><aff id="aff2"><label>b</label>Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673, Singapore</aff><author-notes><corresp id="cor1"><label>⁎</label>Corresponding author. Fax: +65 67791117. <email>dxliu@imcb.a-star.edu.sg</email></corresp></author-notes><pub-date pub-type="pmc-release"><day>4</day><month>5</month><year>2006</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"><day>29</day><month>5</month><year>2006</year></pub-date><pub-date pub-type="epub"><day>4</day><month>5</month><year>2006</year></pub-date><volume>580</volume><issue>13</issue><fpage>3192</fpage><lpage>3200</lpage><history><date date-type="received"><day>3</day><month>3</month><year>2006</year></date><date date-type="rev-recd"><day>18</day><month>4</month><year>2006</year></date><date date-type="accepted"><day>26</day><month>4</month><year>2006</year></date></history><permissions><copyright-statement>Copyright © 2006 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.</copyright-statement><copyright-year>2006</copyright-year><copyright-holder>Federation of European Biochemical Societies</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><p>Coronavirus envelope (E) protein is a small integral membrane protein with multi-functions in virion assembly, morphogenesis and virus–host interaction. Different coronavirus E proteins share striking similarities in biochemical properties and biological functions, but seem to adopt distinct membrane topology. In this report, we study the membrane topology of the SARS-CoV E protein by immunofluorescent staining of cells differentially permeabilized with detergents and proteinase K protection assay. It was revealed that both the N- and C-termini of the SARS-CoV E protein are exposed to the cytoplasmic side of the membranes (N<sub>cyto</sub>C<sub>cyto</sub>). In contrast, parallel experiments showed that the E protein from infectious bronchitis virus (IBV) spanned the membranes once, with the N-terminus exposed luminally and the C-terminus exposed cytoplasmically (N<sub>exo(lum)</sub>C<sub>cyto</sub>). Intriguingly, a minor proportion of the SARS-CoV E protein was found to be modified by N-linked glycosylation on Asn 66 and inserted into the membranes once with the C-terminus exposed to the luminal side. The presence of two distinct membrane topologies of the SARS-CoV E protein may provide a useful clue to the pathogenesis of SARS-CoV.</p></abstract><kwd-group><title>Keywords</title><kwd>SARS-CoV</kwd><kwd>Envelope protein</kwd><kwd>Membrane topology</kwd></kwd-group></article-meta></front></pmc><affiliations><list><country><li>Singapour</li></country></list><tree><country name="Singapour"><noRegion><name sortKey="Yuan, Q" sort="Yuan, Q" uniqKey="Yuan Q" first="Q." last="Yuan">Q. Yuan</name></noRegion><name sortKey="Liao, Y" sort="Liao, Y" uniqKey="Liao Y" first="Y." last="Liao">Y. Liao</name><name sortKey="Liu, D X" sort="Liu, D X" uniqKey="Liu D" first="D. X." last="Liu">D. X. Liu</name><name sortKey="Liu, D X" sort="Liu, D X" uniqKey="Liu D" first="D. X." last="Liu">D. X. Liu</name><name sortKey="Tam, J P" sort="Tam, J P" uniqKey="Tam J" first="J. P." last="Tam">J. P. Tam</name><name sortKey="Torres, J" sort="Torres, J" uniqKey="Torres J" first="J." last="Torres">J. Torres</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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