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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus<xref ref-type="fn" rid="FN1">*</xref><xref ref-type="fn" rid="FN2"><sup><inline-graphic xlink:href="sbox.jpg"></inline-graphic></sup></xref></title><author><name sortKey="Milbradt, Jens" sort="Milbradt, Jens" uniqKey="Milbradt J" first="Jens" last="Milbradt">Jens Milbradt</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kraut, Alexandra" sort="Kraut, Alexandra" uniqKey="Kraut A" first="Alexandra" last="Kraut">Alexandra Kraut</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Hutterer, Corina" sort="Hutterer, Corina" uniqKey="Hutterer C" first="Corina" last="Hutterer">Corina Hutterer</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Sonntag, Eric" sort="Sonntag, Eric" uniqKey="Sonntag E" first="Eric" last="Sonntag">Eric Sonntag</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Schmeiser, Cathrin" sort="Schmeiser, Cathrin" uniqKey="Schmeiser C" first="Cathrin" last="Schmeiser">Cathrin Schmeiser</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ferro, Myriam" sort="Ferro, Myriam" uniqKey="Ferro M" first="Myriam" last="Ferro">Myriam Ferro</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Wagner, Sabrina" sort="Wagner, Sabrina" uniqKey="Wagner S" first="Sabrina" last="Wagner">Sabrina Wagner</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lenac, Tihana" sort="Lenac, Tihana" uniqKey="Lenac T" first="Tihana" last="Lenac">Tihana Lenac</name><affiliation><nlm:aff id="aff5">**Department of Histology and Embryology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia;</nlm:aff></affiliation></author><author><name sortKey="Claus, Claudia" sort="Claus, Claudia" uniqKey="Claus C" first="Claudia" last="Claus">Claudia Claus</name><affiliation><nlm:aff id="aff6">‡‡Institute for Virology, University of Leipzig, 04103 Leipzig, Germany;</nlm:aff></affiliation></author><author><name sortKey="Pinkert, Sandra" sort="Pinkert, Sandra" uniqKey="Pinkert S" first="Sandra" last="Pinkert">Sandra Pinkert</name><affiliation><nlm:aff id="aff7">§§Institute of Biotechnology, University of Technology Berlin, 13353 Berlin, Germany;</nlm:aff></affiliation></author><author><name sortKey="Hamilton, Stuart T" sort="Hamilton, Stuart T" uniqKey="Hamilton S" first="Stuart T." last="Hamilton">Stuart T. Hamilton</name><affiliation><nlm:aff id="aff8">¶¶Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, 2052 Sydney, Australia;</nlm:aff></affiliation></author><author><name sortKey="Rawlinson, William D" sort="Rawlinson, William D" uniqKey="Rawlinson W" first="William D." last="Rawlinson">William D. Rawlinson</name><affiliation><nlm:aff id="aff8">¶¶Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, 2052 Sydney, Australia;</nlm:aff></affiliation></author><author><name sortKey="Sticht, Heinrich" sort="Sticht, Heinrich" uniqKey="Sticht H" first="Heinrich" last="Sticht">Heinrich Sticht</name><affiliation><nlm:aff id="aff9">‖‖Division of Bioinformatics, Institute of Biochemistry, University of Erlangen-Nuremberg, 91054 Erlangen, Germany</nlm:aff></affiliation></author><author><name sortKey="Coute, Yohann" sort="Coute, Yohann" uniqKey="Coute Y" first="Yohann" last="Couté">Yohann Couté</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Marschall, Manfred" sort="Marschall, Manfred" uniqKey="Marschall M" first="Manfred" last="Marschall">Manfred Marschall</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">24969177</idno><idno type="pmc">4125742</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4125742</idno><idno type="RBID">PMC:4125742</idno><idno type="doi">10.1074/mcp.M113.035782</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">001A27</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">001A27</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus<xref ref-type="fn" rid="FN1">*</xref><xref ref-type="fn" rid="FN2"><sup><inline-graphic xlink:href="sbox.jpg"></inline-graphic></sup></xref></title><author><name sortKey="Milbradt, Jens" sort="Milbradt, Jens" uniqKey="Milbradt J" first="Jens" last="Milbradt">Jens Milbradt</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Kraut, Alexandra" sort="Kraut, Alexandra" uniqKey="Kraut A" first="Alexandra" last="Kraut">Alexandra Kraut</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Hutterer, Corina" sort="Hutterer, Corina" uniqKey="Hutterer C" first="Corina" last="Hutterer">Corina Hutterer</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Sonntag, Eric" sort="Sonntag, Eric" uniqKey="Sonntag E" first="Eric" last="Sonntag">Eric Sonntag</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Schmeiser, Cathrin" sort="Schmeiser, Cathrin" uniqKey="Schmeiser C" first="Cathrin" last="Schmeiser">Cathrin Schmeiser</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Ferro, Myriam" sort="Ferro, Myriam" uniqKey="Ferro M" first="Myriam" last="Ferro">Myriam Ferro</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Wagner, Sabrina" sort="Wagner, Sabrina" uniqKey="Wagner S" first="Sabrina" last="Wagner">Sabrina Wagner</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author><author><name sortKey="Lenac, Tihana" sort="Lenac, Tihana" uniqKey="Lenac T" first="Tihana" last="Lenac">Tihana Lenac</name><affiliation><nlm:aff id="aff5">**Department of Histology and Embryology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia;</nlm:aff></affiliation></author><author><name sortKey="Claus, Claudia" sort="Claus, Claudia" uniqKey="Claus C" first="Claudia" last="Claus">Claudia Claus</name><affiliation><nlm:aff id="aff6">‡‡Institute for Virology, University of Leipzig, 04103 Leipzig, Germany;</nlm:aff></affiliation></author><author><name sortKey="Pinkert, Sandra" sort="Pinkert, Sandra" uniqKey="Pinkert S" first="Sandra" last="Pinkert">Sandra Pinkert</name><affiliation><nlm:aff id="aff7">§§Institute of Biotechnology, University of Technology Berlin, 13353 Berlin, Germany;</nlm:aff></affiliation></author><author><name sortKey="Hamilton, Stuart T" sort="Hamilton, Stuart T" uniqKey="Hamilton S" first="Stuart T." last="Hamilton">Stuart T. Hamilton</name><affiliation><nlm:aff id="aff8">¶¶Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, 2052 Sydney, Australia;</nlm:aff></affiliation></author><author><name sortKey="Rawlinson, William D" sort="Rawlinson, William D" uniqKey="Rawlinson W" first="William D." last="Rawlinson">William D. Rawlinson</name><affiliation><nlm:aff id="aff8">¶¶Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, 2052 Sydney, Australia;</nlm:aff></affiliation></author><author><name sortKey="Sticht, Heinrich" sort="Sticht, Heinrich" uniqKey="Sticht H" first="Heinrich" last="Sticht">Heinrich Sticht</name><affiliation><nlm:aff id="aff9">‖‖Division of Bioinformatics, Institute of Biochemistry, University of Erlangen-Nuremberg, 91054 Erlangen, Germany</nlm:aff></affiliation></author><author><name sortKey="Coute, Yohann" sort="Coute, Yohann" uniqKey="Coute Y" first="Yohann" last="Couté">Yohann Couté</name><affiliation><nlm:aff id="aff2">Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</nlm:aff></affiliation><affiliation><nlm:aff id="aff4">INSERM, BGE, F-38000 Grenoble, France;</nlm:aff></affiliation></author><author><name sortKey="Marschall, Manfred" sort="Marschall, Manfred" uniqKey="Marschall M" first="Manfred" last="Marschall">Manfred Marschall</name><affiliation><nlm:aff id="aff1"></nlm:aff></affiliation></author></analytic><series><title level="j">Molecular & Cellular Proteomics : MCP</title><idno type="ISSN">1535-9476</idno><idno type="eISSN">1535-9484</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Herpesviral capsids are assembled in the host cell nucleus before being translocated into the cytoplasm for further maturation. The crossing of the nuclear envelope represents a major event that requires the formation of the nuclear egress complex (NEC). Previous studies demonstrated that human cytomegalovirus (HCMV) proteins pUL50 and pUL53, as well as their homologs in all members of <italic>Herpesviridae</italic>, interact with each other at the nuclear envelope and form the heterodimeric core of the NEC. In order to characterize further the viral and cellular protein content of the multimeric NEC, the native complex was isolated from HCMV-infected human primary fibroblasts at various time points and analyzed using quantitative proteomics. Previously postulated components of the HCMV-specific NEC, as well as novel potential NEC-associated proteins such as emerin, were identified. In this regard, interaction and colocalization between emerin and pUL50 were confirmed by coimmunoprecipitation and confocal microscopy analyses, respectively. A functional validation of viral and cellular NEC constituents was achieved through siRNA-mediated knockdown experiments. The important role of emerin in NEC functionality was demonstrated by a reduction of viral replication when emerin expression was down-regulated. Moreover, under such conditions, reduced production of viral proteins and deregulation of viral late cytoplasmic maturation were observed. Combined, these data prove the functional importance of emerin as an NEC component, associated with pUL50, pUL53, pUL97, p32/gC1qR, and further regulatory proteins. Summarized, our findings provide the first proteomics-based characterization and functional validation of the HCMV-specific multimeric NEC.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Mol Cell Proteomics</journal-id><journal-id journal-id-type="iso-abbrev">Mol. Cell Proteomics</journal-id><journal-id journal-id-type="hwp">mcprot</journal-id><journal-id journal-id-type="pmc">mcprot</journal-id><journal-id journal-id-type="publisher-id">MCP</journal-id><journal-title-group><journal-title>Molecular & Cellular Proteomics : MCP</journal-title></journal-title-group><issn pub-type="ppub">1535-9476</issn><issn pub-type="epub">1535-9484</issn><publisher><publisher-name>The American Society for Biochemistry and Molecular Biology</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">24969177</article-id><article-id pub-id-type="pmc">4125742</article-id><article-id pub-id-type="publisher-id">M113.035782</article-id><article-id pub-id-type="doi">10.1074/mcp.M113.035782</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group></article-categories><title-group><article-title>Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus<xref ref-type="fn" rid="FN1">*</xref><xref ref-type="fn" rid="FN2"><sup><inline-graphic xlink:href="sbox.jpg"></inline-graphic></sup></xref></article-title><alt-title alt-title-type="short">Proteins of the HCMV-specific Nuclear Egress Complex</alt-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Milbradt</surname><given-names>Jens</given-names></name><xref ref-type="aff" rid="aff1">‡</xref></contrib><contrib contrib-type="author"><name><surname>Kraut</surname><given-names>Alexandra</given-names></name><xref ref-type="aff" rid="aff2">§</xref><xref ref-type="aff" rid="aff3">¶</xref><xref ref-type="aff" rid="aff4">‖</xref></contrib><contrib contrib-type="author"><name><surname>Hutterer</surname><given-names>Corina</given-names></name><xref ref-type="aff" rid="aff1">‡</xref></contrib><contrib contrib-type="author"><name><surname>Sonntag</surname><given-names>Eric</given-names></name><xref ref-type="aff" rid="aff1">‡</xref></contrib><contrib contrib-type="author"><name><surname>Schmeiser</surname><given-names>Cathrin</given-names></name><xref ref-type="aff" rid="aff1">‡</xref></contrib><contrib contrib-type="author"><name><surname>Ferro</surname><given-names>Myriam</given-names></name><xref ref-type="aff" rid="aff2">§</xref><xref ref-type="aff" rid="aff3">¶</xref><xref ref-type="aff" rid="aff4">‖</xref></contrib><contrib contrib-type="author"><name><surname>Wagner</surname><given-names>Sabrina</given-names></name><xref ref-type="aff" rid="aff1">‡</xref></contrib><contrib contrib-type="author"><name><surname>Lenac</surname><given-names>Tihana</given-names></name><xref ref-type="aff" rid="aff5">**</xref></contrib><contrib contrib-type="author"><name><surname>Claus</surname><given-names>Claudia</given-names></name><xref ref-type="aff" rid="aff6">‡‡</xref></contrib><contrib contrib-type="author"><name><surname>Pinkert</surname><given-names>Sandra</given-names></name><xref ref-type="aff" rid="aff7">§§</xref></contrib><contrib contrib-type="author"><name><surname>Hamilton</surname><given-names>Stuart T.</given-names></name><xref ref-type="aff" rid="aff8">¶¶</xref></contrib><contrib contrib-type="author"><name><surname>Rawlinson</surname><given-names>William D.</given-names></name><xref ref-type="aff" rid="aff8">¶¶</xref></contrib><contrib contrib-type="author"><name><surname>Sticht</surname><given-names>Heinrich</given-names></name><xref ref-type="aff" rid="aff9">‖‖</xref></contrib><contrib contrib-type="author"><name><surname>Couté</surname><given-names>Yohann</given-names></name><xref ref-type="aff" rid="aff2">§</xref><xref ref-type="aff" rid="aff3">¶</xref><xref ref-type="aff" rid="aff4">‖</xref><xref ref-type="corresp" rid="cor1"><italic><sup>a</sup></italic></xref></contrib><contrib contrib-type="author"><name><surname>Marschall</surname><given-names>Manfred</given-names></name><xref ref-type="aff" rid="aff1">‡</xref><xref ref-type="corresp" rid="cor1"><italic><sup>a</sup></italic></xref></contrib><aff id="aff1">From the<sup>‡</sup>Institute for Clinical and Molecular Virology, University of Erlangen-Nuremberg, 91054 Erlangen, Germany;</aff><aff id="aff2"><sup>§</sup>Université Grenoble Alpes, iRTSV-BGE, F-38000 Grenoble, France;</aff><aff id="aff3">¶CEA, iRTSV-BGE, F-38000 Grenoble, France;</aff><aff id="aff4"><sup>‖</sup>INSERM, BGE, F-38000 Grenoble, France;</aff><aff id="aff5">**Department of Histology and Embryology, Faculty of Medicine, University of Rijeka, 51000 Rijeka, Croatia;</aff><aff id="aff6">‡‡Institute for Virology, University of Leipzig, 04103 Leipzig, Germany;</aff><aff id="aff7">§§Institute of Biotechnology, University of Technology Berlin, 13353 Berlin, Germany;</aff><aff id="aff8">¶¶Virology Division, SEALS Microbiology, Prince of Wales Hospital, University of New South Wales, 2052 Sydney, Australia;</aff><aff id="aff9">‖‖Division of Bioinformatics, Institute of Biochemistry, University of Erlangen-Nuremberg, 91054 Erlangen, Germany</aff></contrib-group><author-notes><corresp id="cor1"><italic><sup>a</sup></italic> To whom correspondence should be addressed: Manfred Marschall, Tel.: <phone>49-0-9131-8526089</phone>, E-mail: <email>manfred.marschall@viro.med.uni-erlangen.de</email>; Yohann Couté, Tel.: <phone>33-0-438789461</phone>, E-mail: <email>yohann.coute@cea.fr</email>.</corresp></author-notes><pub-date pub-type="ppub"><month>8</month><year>2014</year></pub-date><pub-date pub-type="epub"><day>26</day><month>6</month><year>2014</year></pub-date><pub-date pub-type="pmc-release"><day>1</day><month>8</month><year>2015</year></pub-date><pmc-comment> PMC Release delay is 12 months and 0 days and was based on the
<volume>13</volume><issue>8</issue><fpage>2132</fpage><lpage>2146</lpage><history><date date-type="received"><day>31</day><month>10</month><year>2013</year></date><date date-type="rev-recd"><day>7</day><month>5</month><year>2014</year></date></history><permissions><copyright-statement>© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.</copyright-statement><copyright-year>2014</copyright-year></permissions><self-uri xlink:title="pdf" xlink:type="simple" xlink:href="zjw00814002132.pdf"></self-uri><abstract><p>Herpesviral capsids are assembled in the host cell nucleus before being translocated into the cytoplasm for further maturation. The crossing of the nuclear envelope represents a major event that requires the formation of the nuclear egress complex (NEC). Previous studies demonstrated that human cytomegalovirus (HCMV) proteins pUL50 and pUL53, as well as their homologs in all members of <italic>Herpesviridae</italic>, interact with each other at the nuclear envelope and form the heterodimeric core of the NEC. In order to characterize further the viral and cellular protein content of the multimeric NEC, the native complex was isolated from HCMV-infected human primary fibroblasts at various time points and analyzed using quantitative proteomics. Previously postulated components of the HCMV-specific NEC, as well as novel potential NEC-associated proteins such as emerin, were identified. In this regard, interaction and colocalization between emerin and pUL50 were confirmed by coimmunoprecipitation and confocal microscopy analyses, respectively. A functional validation of viral and cellular NEC constituents was achieved through siRNA-mediated knockdown experiments. The important role of emerin in NEC functionality was demonstrated by a reduction of viral replication when emerin expression was down-regulated. Moreover, under such conditions, reduced production of viral proteins and deregulation of viral late cytoplasmic maturation were observed. Combined, these data prove the functional importance of emerin as an NEC component, associated with pUL50, pUL53, pUL97, p32/gC1qR, and further regulatory proteins. Summarized, our findings provide the first proteomics-based characterization and functional validation of the HCMV-specific multimeric NEC.</p></abstract></article-meta></front></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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