Structural characterization of recombinant IAV polymerase reveals a stable complex between viral PA-PB1 heterodimer and host RanBP5.
Identifieur interne : 000065 ( France/Analysis ); précédent : 000064; suivant : 000066Structural characterization of recombinant IAV polymerase reveals a stable complex between viral PA-PB1 heterodimer and host RanBP5.
Auteurs : Christopher Swale [France] ; Alexandre Monod [France] ; Laura Tengo [France] ; Alice Labaronne [France] ; Frédéric Garzoni [France] ; Jean-Marie Bourhis [France] ; Stephen Cusack [France] ; Guy Schoehn [France] ; Imre Berger [France] ; Rob W H. Ruigrok [France] ; Thibaut Crépin [France]Source :
- Scientific Reports [ 2045-2322 ] ; 2016-04-20.
Abstract
The genome of influenza A virus (IAV) comprises eight RNA segments (vRNA) which are transcribed and replicated by the heterotrimeric IAV RNA-dependent RNA-polymerase (RdRp). RdRp consists of three subunits (PA, PB1 and PB2) and binds both the highly conserved 3'- and 5'-ends of the vRNA segment. The IAV RdRp is an important antiviral target, but its structural mechanism has remained largely elusive to date. By applying a polyprotein strategy, we produced RdRp complexes and define a minimal human IAV RdRp core complex. We show that PA-PB1 forms a stable heterodimeric submodule that can strongly interact with 5'-vRNA. In contrast, 3'-vRNA recognition critically depends on the PB2 N-terminal domain. Moreover, we demonstrate that PA-PB1 forms a stable and stoichiometric complex with host nuclear import factor RanBP5 that can be modelled using SAXS and we show that the PA-PB1-RanPB5 complex is no longer capable of 5'-vRNA binding. Our results provide further evidence for a step-wise assembly of IAV structural components, regulated by nuclear transport mechanisms and host factor binding.
Url:
DOI: 10.1038/srep24727
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream Hal, to step Corpus: 000268
- to stream Hal, to step Curation: 000268
- to stream Hal, to step Checkpoint: 000089
- to stream Main, to step Merge: 000381
- to stream Main, to step Curation: 000381
- to stream Main, to step Exploration: 000381
- to stream France, to step Extraction: 000065
Links to Exploration step
Hal:hal-01342969Le document en format XML
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38058 Grenoble cedex</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.univ-grenoble-alpes.fr</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country><placeName><settlement type="city">Grenoble</settlement><region type="region" nuts="2">Auvergne-Rhône-Alpes</region><region type="old region" nuts="2">Rhône-Alpes</region></placeName><orgName type="university">Université Grenoble-Alpes</orgName></affiliation></author><author><name sortKey="Berger, Imre" sort="Berger, Imre" uniqKey="Berger I" first="Imre" last="Berger">Imre Berger</name><affiliation wicri:level="1"><hal:affiliation type="institution" xml:id="struct-301930" status="VALID"> <orgName>European Molecular Biology Laboratory [Grenoble]</orgName> <orgName type="acronym">EMBL</orgName> <desc> <address> <addrLine>71 avenue des MartyrsCS 9018138042 Grenoble Cedex 9</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.embl.fr/</ref> </desc> </hal:affiliation><country>France</country></affiliation></author><author><name sortKey="Ruigrok, Rob W H" sort="Ruigrok, Rob W H" uniqKey="Ruigrok R" first="Rob W H" last="Ruigrok">Rob W H. Ruigrok</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-576" status="VALID"> <idno type="IdRef">06912633X</idno> <idno type="ISNI">0000 0004 0641 5776</idno> <idno type="RNSR">199911764W</idno> <orgName>Institut de biologie structurale</orgName> <orgName type="acronym">IBS - UMR 5075</orgName> <date type="start">1992-01-01</date> <desc> <address> <addrLine>Institut de Biologie Structurale, 71 avenue des martyrs, CS 10090, 38044 Grenoble Cedex 9</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.ibs.fr/</ref> </desc> <listRelation> <relation name="UMR5075" active="#struct-441569" type="direct"></relation> <relation name="DRF/IRIG/DBSC" active="#struct-555292" type="direct"></relation> <relation name="DRF/IRIG" active="#struct-465913" type="indirect"></relation> <relation active="#struct-300016" type="indirect"></relation> <relation active="#struct-445543" type="direct"></relation> </listRelation> <tutelles><tutelle name="UMR5075" active="#struct-441569" type="direct"><org type="institution" xml:id="struct-441569" status="VALID"> <idno type="IdRef">02636817X</idno> <idno type="ISNI">0000000122597504</idno> <orgName>Centre National de la Recherche Scientifique</orgName> <orgName type="acronym">CNRS</orgName> <date type="start">1939-10-19</date> <desc> <address> <country key="FR"></country> </address> <ref type="url">http://www.cnrs.fr/</ref> </desc> </org></tutelle><tutelle name="DRF/IRIG/DBSC" active="#struct-555292" type="direct"><org type="laboratory" xml:id="struct-555292" status="VALID"> <orgName>Institut de Recherche Interdisciplinaire de Grenoble</orgName> <orgName type="acronym">IRIG</orgName> <date type="start">2019-01-01</date> <desc> <address> <country key="FR"></country> </address> <ref type="url">https://hal-cea.archives-ouvertes.fr/IRIG/</ref> </desc> <listRelation> <relation name="DRF/IRIG" active="#struct-465913" type="direct"></relation> <relation active="#struct-300016" type="indirect"></relation> </listRelation> </org></tutelle><tutelle name="DRF/IRIG" active="#struct-465913" type="indirect"><org type="regrouplaboratory" xml:id="struct-465913" status="VALID"> <idno type="RNSR">201622496M</idno> <orgName>Direction de Recherche Fondamentale (CEA)</orgName> <orgName type="acronym">DRF (CEA)</orgName> <date type="start">2016-01-01</date> <desc> <address> <addrLine>Direction de Recherche FondamentaleFundamental Research DivisionCEA Saclay91191 Gif-sur-Yvette</addrLine> <country key="FR"></country> </address> <ref type="url">http://ceasciences.fr/</ref> </desc> <listRelation> <relation active="#struct-300016" type="direct"></relation> </listRelation> </org></tutelle><tutelle active="#struct-300016" type="indirect"><org type="institution" xml:id="struct-300016" status="VALID"> <orgName>Commissariat à l'énergie atomique et aux énergies alternatives</orgName> <orgName type="acronym">CEA</orgName> <desc> <address> <addrLine>Centre de SaclayCentre de GrenobleCentre de Cadaracheetc</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.cea.fr/</ref> </desc> </org></tutelle><tutelle active="#struct-445543" type="direct"><org type="institution" xml:id="struct-445543" status="VALID"> <idno type="IdRef">188399275</idno> <orgName>Université Grenoble Alpes</orgName> <orgName type="acronym">UGA</orgName> <date type="start">2016-01-01</date> <desc> <address> <addrLine>CS 40700 - 38058 Grenoble cedex</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.univ-grenoble-alpes.fr</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country><placeName><settlement type="city">Grenoble</settlement><region type="region" nuts="2">Auvergne-Rhône-Alpes</region><region type="old region" nuts="2">Rhône-Alpes</region></placeName><orgName type="university">Université Grenoble-Alpes</orgName></affiliation></author><author><name sortKey="Crepin, Thibaut" sort="Crepin, Thibaut" uniqKey="Crepin T" first="Thibaut" last="Crépin">Thibaut Crépin</name><affiliation wicri:level="1"><hal:affiliation type="department" xml:id="struct-576" status="VALID"> <idno type="IdRef">06912633X</idno> <idno type="ISNI">0000 0004 0641 5776</idno> <idno type="RNSR">199911764W</idno> <orgName>Institut de biologie structurale</orgName> <orgName type="acronym">IBS - UMR 5075</orgName> <date type="start">1992-01-01</date> <desc> <address> <addrLine>Institut de Biologie Structurale, 71 avenue des martyrs, CS 10090, 38044 Grenoble Cedex 9</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.ibs.fr/</ref> </desc> <listRelation> <relation name="UMR5075" active="#struct-441569" type="direct"></relation> <relation name="DRF/IRIG/DBSC" active="#struct-555292" type="direct"></relation> <relation name="DRF/IRIG" active="#struct-465913" type="indirect"></relation> <relation active="#struct-300016" type="indirect"></relation> <relation active="#struct-445543" type="direct"></relation> </listRelation> <tutelles><tutelle name="UMR5075" active="#struct-441569" type="direct"><org type="institution" xml:id="struct-441569" status="VALID"> <idno type="IdRef">02636817X</idno> <idno type="ISNI">0000000122597504</idno> <orgName>Centre National de la Recherche Scientifique</orgName> <orgName type="acronym">CNRS</orgName> <date type="start">1939-10-19</date> <desc> <address> <country key="FR"></country> </address> <ref type="url">http://www.cnrs.fr/</ref> </desc> </org></tutelle><tutelle name="DRF/IRIG/DBSC" active="#struct-555292" type="direct"><org type="laboratory" xml:id="struct-555292" status="VALID"> <orgName>Institut de Recherche Interdisciplinaire de Grenoble</orgName> <orgName type="acronym">IRIG</orgName> <date type="start">2019-01-01</date> <desc> <address> <country key="FR"></country> </address> <ref type="url">https://hal-cea.archives-ouvertes.fr/IRIG/</ref> </desc> <listRelation> <relation name="DRF/IRIG" active="#struct-465913" type="direct"></relation> <relation active="#struct-300016" type="indirect"></relation> </listRelation> </org></tutelle><tutelle name="DRF/IRIG" active="#struct-465913" type="indirect"><org type="regrouplaboratory" xml:id="struct-465913" status="VALID"> <idno type="RNSR">201622496M</idno> <orgName>Direction de Recherche Fondamentale (CEA)</orgName> <orgName type="acronym">DRF (CEA)</orgName> <date type="start">2016-01-01</date> <desc> <address> <addrLine>Direction de Recherche FondamentaleFundamental Research DivisionCEA Saclay91191 Gif-sur-Yvette</addrLine> <country key="FR"></country> </address> <ref type="url">http://ceasciences.fr/</ref> </desc> <listRelation> <relation active="#struct-300016" type="direct"></relation> </listRelation> </org></tutelle><tutelle active="#struct-300016" type="indirect"><org type="institution" xml:id="struct-300016" status="VALID"> <orgName>Commissariat à l'énergie atomique et aux énergies alternatives</orgName> <orgName type="acronym">CEA</orgName> <desc> <address> <addrLine>Centre de SaclayCentre de GrenobleCentre de Cadaracheetc</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.cea.fr/</ref> </desc> </org></tutelle><tutelle active="#struct-445543" type="direct"><org type="institution" xml:id="struct-445543" status="VALID"> <idno type="IdRef">188399275</idno> <orgName>Université Grenoble Alpes</orgName> <orgName type="acronym">UGA</orgName> <date type="start">2016-01-01</date> <desc> <address> <addrLine>CS 40700 - 38058 Grenoble cedex</addrLine> <country key="FR"></country> </address> <ref type="url">http://www.univ-grenoble-alpes.fr</ref> </desc> </org></tutelle></tutelles></hal:affiliation><country>France</country><placeName><settlement type="city">Grenoble</settlement><region type="region" nuts="2">Auvergne-Rhône-Alpes</region><region type="old region" nuts="2">Rhône-Alpes</region></placeName><orgName type="university">Université Grenoble-Alpes</orgName></affiliation></author></analytic><idno type="DOI">10.1038/srep24727</idno><series><title level="j">Scientific Reports</title><idno type="ISSN">2045-2322</idno><imprint><date type="datePub">2016-04-20</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"> <p>The genome of influenza A virus (IAV) comprises eight RNA segments (vRNA) which are transcribed and replicated by the heterotrimeric IAV RNA-dependent RNA-polymerase (RdRp). RdRp consists of three subunits (PA, PB1 and PB2) and binds both the highly conserved 3'- and 5'-ends of the vRNA segment. The IAV RdRp is an important antiviral target, but its structural mechanism has remained largely elusive to date. By applying a polyprotein strategy, we produced RdRp complexes and define a minimal human IAV RdRp core complex. We show that PA-PB1 forms a stable heterodimeric submodule that can strongly interact with 5'-vRNA. In contrast, 3'-vRNA recognition critically depends on the PB2 N-terminal domain. Moreover, we demonstrate that PA-PB1 forms a stable and stoichiometric complex with host nuclear import factor RanBP5 that can be modelled using SAXS and we show that the PA-PB1-RanPB5 complex is no longer capable of 5'-vRNA binding. Our results provide further evidence for a step-wise assembly of IAV structural components, regulated by nuclear transport mechanisms and host factor binding.</p> </div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Auvergne-Rhône-Alpes</li><li>Rhône-Alpes</li></region><settlement><li>Grenoble</li></settlement><orgName><li>Université Grenoble-Alpes</li></orgName></list><tree><country name="France"><region name="Auvergne-Rhône-Alpes"><name sortKey="Swale, Christopher" sort="Swale, Christopher" uniqKey="Swale C" first="Christopher" last="Swale">Christopher Swale</name></region><name sortKey="Berger, Imre" sort="Berger, Imre" uniqKey="Berger I" first="Imre" last="Berger">Imre Berger</name><name sortKey="Bourhis, Jean Marie" sort="Bourhis, Jean Marie" uniqKey="Bourhis J" first="Jean-Marie" last="Bourhis">Jean-Marie Bourhis</name><name sortKey="Crepin, Thibaut" sort="Crepin, Thibaut" uniqKey="Crepin T" first="Thibaut" last="Crépin">Thibaut Crépin</name><name sortKey="Cusack, Stephen" sort="Cusack, Stephen" uniqKey="Cusack S" first="Stephen" last="Cusack">Stephen Cusack</name><name sortKey="Garzoni, Frederic" sort="Garzoni, Frederic" uniqKey="Garzoni F" first="Frédéric" last="Garzoni">Frédéric Garzoni</name><name sortKey="Labaronne, Alice" sort="Labaronne, Alice" uniqKey="Labaronne A" first="Alice" last="Labaronne">Alice Labaronne</name><name sortKey="Monod, Alexandre" sort="Monod, Alexandre" uniqKey="Monod A" first="Alexandre" last="Monod">Alexandre Monod</name><name sortKey="Ruigrok, Rob W H" sort="Ruigrok, Rob W H" uniqKey="Ruigrok R" first="Rob W H" last="Ruigrok">Rob W H. Ruigrok</name><name sortKey="Schoehn, Guy" sort="Schoehn, Guy" uniqKey="Schoehn G" first="Guy" last="Schoehn">Guy Schoehn</name><name sortKey="Tengo, Laura" sort="Tengo, Laura" uniqKey="Tengo L" first="Laura" last="Tengo">Laura Tengo</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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