Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling
Identifieur interne : 002073 ( Ncbi/Curation ); précédent : 002072; suivant : 002074Determination of host proteins composing the microenvironment of coronavirus replicase complexes by proximity-labeling
Auteurs : Philip V'Kovski [Suisse] ; Markus Gerber [Suisse] ; Jenna Kelly [Suisse] ; Stephanie Pfaender [Suisse] ; Nadine Ebert [Suisse] ; Sophie Braga Lagache [Suisse] ; Cedric Simillion [Suisse] ; Jasmine Portmann [Suisse] ; Hanspeter Stalder [Suisse] ; Véronique Gaschen [Suisse] ; Rémy Bruggmann [Suisse] ; Michael H. Stoffel [Suisse] ; Manfred Heller [Suisse] ; Ronald Dijkman [Suisse] ; Volker Thiel [Suisse]Source :
- eLife [ 2050-084X ] ; ????.
Abstract
Positive-sense RNA viruses hijack intracellular membranes that provide niches for viral RNA synthesis and a platform for interactions with host proteins. However, little is known about host factors at the interface between replicase complexes and the host cytoplasm. We engineered a biotin ligase into a coronaviral replication/transcription complex (RTC) and identified >500 host proteins constituting the RTC microenvironment. siRNA-silencing of each RTC-proximal host factor demonstrated importance of vesicular trafficking pathways, ubiquitin-dependent and autophagy-related processes, and translation initiation factors. Notably, detection of translation initiation factors at the RTC was instrumental to visualize and demonstrate active translation proximal to replication complexes of several coronaviruses. Collectively, we establish a spatial link between viral RNA synthesis and diverse host factors of unprecedented breadth. Our data may serve as a paradigm for other positive-strand RNA viruses and provide a starting point for a comprehensive analysis of critical virus-host interactions that represent targets for therapeutic intervention.
Url:
DOI: 10.7554/eLife.42037
PubMed: 30632963
PubMed Central: 6372286
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PMC:6372286Le document en format XML
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<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Institute of Virology and Immunology IVI</institution>
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<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution content-type="dept">Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Gaschen, Veronique" sort="Gaschen, Veronique" uniqKey="Gaschen V" first="Véronique" last="Gaschen">Véronique Gaschen</name>
<affiliation wicri:level="1"><nlm:aff id="aff7"><institution content-type="dept">Division of Veterinary Anatomy, Vetsuisse Faculty</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Bruggmann, Remy" sort="Bruggmann, Remy" uniqKey="Bruggmann R" first="Rémy" last="Bruggmann">Rémy Bruggmann</name>
<affiliation wicri:level="1"><nlm:aff id="aff4"><institution content-type="dept">Interfaculty Bioinformatics Unit, SIB Swiss Institute of Bioinformatics</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Stoffel, Michael H" sort="Stoffel, Michael H" uniqKey="Stoffel M" first="Michael H" last="Stoffel">Michael H. Stoffel</name>
<affiliation wicri:level="1"><nlm:aff id="aff7"><institution content-type="dept">Division of Veterinary Anatomy, Vetsuisse Faculty</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Heller, Manfred" sort="Heller, Manfred" uniqKey="Heller M" first="Manfred" last="Heller">Manfred Heller</name>
<affiliation wicri:level="1"><nlm:aff id="aff5"><institution content-type="dept">Mass Spectrometry and Proteomics Core Facility, Department for BioMedical Research (DBMR)</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Dijkman, Ronald" sort="Dijkman, Ronald" uniqKey="Dijkman R" first="Ronald" last="Dijkman">Ronald Dijkman</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Institute of Virology and Immunology IVI</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution content-type="dept">Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Thiel, Volker" sort="Thiel, Volker" uniqKey="Thiel V" first="Volker" last="Thiel">Volker Thiel</name>
<affiliation wicri:level="1"><nlm:aff id="aff1"><institution>Institute of Virology and Immunology IVI</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
<affiliation wicri:level="1"><nlm:aff id="aff2"><institution content-type="dept">Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty</institution>
<institution>University of Bern</institution>
<addr-line>Bern</addr-line>
<country>Switzerland</country>
</nlm:aff>
<country xml:lang="fr">Suisse</country>
<wicri:regionArea># see nlm:aff country strict</wicri:regionArea>
</affiliation>
</author>
</analytic>
<series><title level="j">eLife</title>
<idno type="eISSN">2050-084X</idno>
<imprint><date when="????">????</date>
</imprint>
</series>
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<front><div type="abstract" xml:lang="en"><p>Positive-sense RNA viruses hijack intracellular membranes that provide niches for viral RNA synthesis and a platform for interactions with host proteins. However, little is known about host factors at the interface between replicase complexes and the host cytoplasm. We engineered a biotin ligase into a coronaviral replication/transcription complex (RTC) and identified >500 host proteins constituting the RTC microenvironment. siRNA-silencing of each RTC-proximal host factor demonstrated importance of vesicular trafficking pathways, ubiquitin-dependent and autophagy-related processes, and translation initiation factors. Notably, detection of translation initiation factors at the RTC was instrumental to visualize and demonstrate active translation proximal to replication complexes of several coronaviruses. Collectively, we establish a spatial link between viral RNA synthesis and diverse host factors of unprecedented breadth. Our data may serve as a paradigm for other positive-strand RNA viruses and provide a starting point for a comprehensive analysis of critical virus-host interactions that represent targets for therapeutic intervention.</p>
</div>
</front>
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