Localization and membrane topology of coronavirus nonstructural protein 4: Involvement of the early secretory pathway in replication
Identifieur interne : 003B53 ( Main/Exploration ); précédent : 003B52; suivant : 003B54Localization and membrane topology of coronavirus nonstructural protein 4: Involvement of the early secretory pathway in replication
Auteurs : M. Oostra [Pays-Bas] ; E. G. Te Lintelo [Pays-Bas] ; M. Deijs [Pays-Bas] ; M. H. Verheije [Pays-Bas] ; P. J. M. Rottier [Pays-Bas] ; C. A. M. De Haan [Pays-Bas]Source :
- Journal of virology [ 0022-538X ] ; 2007.
Descripteurs français
- KwdFr :
- Animaux, Biologie informatique, Chats, Lignée cellulaire, Membrane cellulaire (enzymologie), Membrane cellulaire (virologie), Protéines recombinantes (analyse), Protéines recombinantes (génétique), Protéines recombinantes (métabolisme), Protéines virales non structurales (analyse), Protéines virales non structurales (génétique), Protéines virales non structurales (métabolisme), Protéines à fluorescence verte (analyse), Protéines à fluorescence verte (génétique), Protéines à fluorescence verte (métabolisme), RNA replicase (analyse), RNA replicase (génétique), RNA replicase (métabolisme), Réplication virale, Réticulum endoplasmique (enzymologie), Souris, Virus de l'hépatite murine (génétique), Virus de l'hépatite murine (métabolisme), Virus du SRAS (enzymologie), Virus du SRAS (physiologie).
- MESH :
- analyse : Protéines recombinantes, Protéines virales non structurales, Protéines à fluorescence verte, RNA replicase.
- enzymologie : Membrane cellulaire, Réticulum endoplasmique, Virus du SRAS.
- génétique : Protéines recombinantes, Protéines virales non structurales, Protéines à fluorescence verte, RNA replicase, Virus de l'hépatite murine.
- métabolisme : Protéines recombinantes, Protéines virales non structurales, Protéines à fluorescence verte, RNA replicase, Virus de l'hépatite murine.
- physiologie : Virus du SRAS.
- virologie : Membrane cellulaire.
- Pascal (Inist)
English descriptors
- KwdEn :
- Animals, Cats, Cell Line, Cell Membrane (enzymology), Cell Membrane (virology), Computational Biology, Coronavirus, Endoplasmic Reticulum (enzymology), Green Fluorescent Proteins (analysis), Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Localization, Membrane protein, Mice, Murine hepatitis virus (genetics), Murine hepatitis virus (metabolism), RNA Replicase (analysis), RNA Replicase (genetics), RNA Replicase (metabolism), Recombinant Proteins (analysis), Recombinant Proteins (genetics), Recombinant Proteins (metabolism), Replication, SARS Virus (enzymology), SARS Virus (physiology), Viral Nonstructural Proteins (analysis), Viral Nonstructural Proteins (genetics), Viral Nonstructural Proteins (metabolism), Virology, Virus Replication.
- MESH :
- chemical , analysis : Green Fluorescent Proteins, RNA Replicase, Recombinant Proteins, Viral Nonstructural Proteins.
- enzymology : Cell Membrane, Endoplasmic Reticulum, SARS Virus.
- chemical , genetics : Green Fluorescent Proteins, Murine hepatitis virus, RNA Replicase, Recombinant Proteins, Viral Nonstructural Proteins.
- chemical , metabolism : Green Fluorescent Proteins, Murine hepatitis virus, RNA Replicase, Recombinant Proteins, Viral Nonstructural Proteins.
- physiology : SARS Virus.
- virology : Cell Membrane.
- Animals, Cats, Cell Line, Computational Biology, Mice, Virus Replication.
Abstract
The coronavirus nonstructural proteins (nsp's) derived from the replicase polyproteins collectively constitute the viral replication complexes, which are anchored to double-membrane vesicles. Little is known about the biogenesis of these complexes, the membrane anchoring of which is probably mediated by nsp3, nsp4, and nsp6, as they contain several putative transmembrane domains. As a first step to getting more insight into the formation of the coronavirus replication complex, the membrane topology, processing, and subcellular localization of nsp4 of the mouse hepatitis virus (MHV) and severe acute respiratory syndrome-associated coronavirus (SARS-CoV) were elucidated in this study. Both nsp4 proteins became N glycosylated, while their amino and carboxy termini were localized to the cytoplasm. These observations imply nsp4 to assemble in the membrane as a tetraspanning transmembrane protein with a Nendo/Cendo topology. The amino terminus of SARS-CoV nsp4, but not that of MHV nsp4, was shown to be (partially) processed by signal peptidase. nsp4 localized to the endoplasmic reticulum (ER) when expressed alone but was recruited to the replication complexes in infected cells, nsp4 present in these complexes did not colocalize with markers of the ER or Golgi apparatus, while the susceptibility of its sugars to endoglycosidase H indicated that the protein had also not traveled trough the latter compartment. The important role of the early secretory pathway in formation of the replication complexes was also demonstrated by the inhibition of coronaviral replication when the ER export machinery was blocked by use of the kinase inhibitor H89 or by expression of a mutant, Sarl[H79G].
Affiliations:
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Le document en format XML
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Oostra</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author><author><name sortKey="Te Lintelo, E G" sort="Te Lintelo, E G" uniqKey="Te Lintelo E" first="E. G." last="Te Lintelo">E. G. Te Lintelo</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author><author><name sortKey="Deijs, M" sort="Deijs, M" uniqKey="Deijs M" first="M." last="Deijs">M. Deijs</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author><author><name sortKey="Verheije, M H" sort="Verheije, M H" uniqKey="Verheije M" first="M. H." last="Verheije">M. H. Verheije</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author><author><name sortKey="Rottier, P J M" sort="Rottier, P J M" uniqKey="Rottier P" first="P. J. M." last="Rottier">P. J. M. Rottier</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author><author><name sortKey="De Haan, C A M" sort="De Haan, C A M" uniqKey="De Haan C" first="C. A. M." last="De Haan">C. A. M. De Haan</name><affiliation wicri:level="4"><inist:fA14 i1="01"><s1>Virology Division, Department of Infectious Diseases and Immunology, Utrecht University, Yalelaan 1</s1><s2>3584 CL Utrecht</s2><s3>NLD</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ><sZ>3 aut.</sZ><sZ>4 aut.</sZ><sZ>5 aut.</sZ><sZ>6 aut.</sZ></inist:fA14><country>Pays-Bas</country><wicri:noRegion>3584 CL Utrecht</wicri:noRegion><orgName type="university">Université d'Utrecht</orgName><placeName><settlement type="city">Utrecht</settlement><region nuts="2">Utrecht (province)</region></placeName></affiliation></author></analytic><series><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno><imprint><date when="2007">2007</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of virology</title><title level="j" type="abbreviated">J. virol.</title><idno type="ISSN">0022-538X</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term><term>Cats</term><term>Cell Line</term><term>Cell Membrane (enzymology)</term><term>Cell Membrane (virology)</term><term>Computational Biology</term><term>Coronavirus</term><term>Endoplasmic Reticulum (enzymology)</term><term>Green Fluorescent Proteins (analysis)</term><term>Green Fluorescent Proteins (genetics)</term><term>Green Fluorescent Proteins (metabolism)</term><term>Localization</term><term>Membrane protein</term><term>Mice</term><term>Murine hepatitis virus (genetics)</term><term>Murine hepatitis virus (metabolism)</term><term>RNA Replicase (analysis)</term><term>RNA Replicase (genetics)</term><term>RNA Replicase (metabolism)</term><term>Recombinant Proteins (analysis)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (metabolism)</term><term>Replication</term><term>SARS Virus (enzymology)</term><term>SARS Virus (physiology)</term><term>Viral Nonstructural Proteins (analysis)</term><term>Viral Nonstructural Proteins (genetics)</term><term>Viral Nonstructural Proteins (metabolism)</term><term>Virology</term><term>Virus Replication</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term><term>Biologie informatique</term><term>Chats</term><term>Lignée cellulaire</term><term>Membrane cellulaire (enzymologie)</term><term>Membrane cellulaire (virologie)</term><term>Protéines recombinantes (analyse)</term><term>Protéines recombinantes (génétique)</term><term>Protéines recombinantes (métabolisme)</term><term>Protéines virales non structurales (analyse)</term><term>Protéines virales non structurales (génétique)</term><term>Protéines virales non structurales (métabolisme)</term><term>Protéines à fluorescence verte (analyse)</term><term>Protéines à fluorescence verte (génétique)</term><term>Protéines à fluorescence verte (métabolisme)</term><term>RNA replicase (analyse)</term><term>RNA replicase (génétique)</term><term>RNA replicase (métabolisme)</term><term>Réplication virale</term><term>Réticulum endoplasmique (enzymologie)</term><term>Souris</term><term>Virus de l'hépatite murine (génétique)</term><term>Virus de l'hépatite murine (métabolisme)</term><term>Virus du SRAS (enzymologie)</term><term>Virus du SRAS (physiologie)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Green Fluorescent Proteins</term><term>RNA Replicase</term><term>Recombinant Proteins</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines virales non structurales</term><term>Protéines à fluorescence verte</term><term>RNA replicase</term></keywords><keywords scheme="MESH" qualifier="enzymologie" xml:lang="fr"><term>Membrane cellulaire</term><term>Réticulum endoplasmique</term><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Cell Membrane</term><term>Endoplasmic Reticulum</term><term>SARS Virus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Murine hepatitis virus</term><term>RNA Replicase</term><term>Recombinant Proteins</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines virales non structurales</term><term>Protéines à fluorescence verte</term><term>RNA replicase</term><term>Virus de l'hépatite murine</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Murine hepatitis virus</term><term>RNA Replicase</term><term>Recombinant Proteins</term><term>Viral Nonstructural Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protéines recombinantes</term><term>Protéines virales non structurales</term><term>Protéines à fluorescence verte</term><term>RNA replicase</term><term>Virus de l'hépatite murine</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Virus du SRAS</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Membrane cellulaire</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Cell Membrane</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cats</term><term>Cell Line</term><term>Computational Biology</term><term>Mice</term><term>Virus Replication</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Animaux</term><term>Biologie informatique</term><term>Chats</term><term>Coronavirus</term><term>Lignée cellulaire</term><term>Localisation</term><term>Protéine membranaire</term><term>Réplication</term><term>Réplication virale</term><term>Souris</term><term>Virologie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The coronavirus nonstructural proteins (nsp's) derived from the replicase polyproteins collectively constitute the viral replication complexes, which are anchored to double-membrane vesicles. Little is known about the biogenesis of these complexes, the membrane anchoring of which is probably mediated by nsp3, nsp4, and nsp6, as they contain several putative transmembrane domains. As a first step to getting more insight into the formation of the coronavirus replication complex, the membrane topology, processing, and subcellular localization of nsp4 of the mouse hepatitis virus (MHV) and severe acute respiratory syndrome-associated coronavirus (SARS-CoV) were elucidated in this study. Both nsp4 proteins became N glycosylated, while their amino and carboxy termini were localized to the cytoplasm. These observations imply nsp4 to assemble in the membrane as a tetraspanning transmembrane protein with a Nendo/Cendo topology. The amino terminus of SARS-CoV nsp4, but not that of MHV nsp4, was shown to be (partially) processed by signal peptidase. nsp4 localized to the endoplasmic reticulum (ER) when expressed alone but was recruited to the replication complexes in infected cells, nsp4 present in these complexes did not colocalize with markers of the ER or Golgi apparatus, while the susceptibility of its sugars to endoglycosidase H indicated that the protein had also not traveled trough the latter compartment. The important role of the early secretory pathway in formation of the replication complexes was also demonstrated by the inhibition of coronaviral replication when the ER export machinery was blocked by use of the kinase inhibitor H89 or by expression of a mutant, Sarl[H79G].</div></front></TEI><affiliations><list><country><li>Pays-Bas</li></country><region><li>Utrecht (province)</li></region><settlement><li>Utrecht</li></settlement><orgName><li>Université d'Utrecht</li></orgName></list><tree><country name="Pays-Bas"><region name="Utrecht (province)"><name sortKey="Oostra, M" sort="Oostra, M" uniqKey="Oostra M" first="M." last="Oostra">M. Oostra</name></region><name sortKey="De Haan, C A M" sort="De Haan, C A M" uniqKey="De Haan C" first="C. A. M." last="De Haan">C. A. M. De Haan</name><name sortKey="Deijs, M" sort="Deijs, M" uniqKey="Deijs M" first="M." last="Deijs">M. Deijs</name><name sortKey="Rottier, P J M" sort="Rottier, P J M" uniqKey="Rottier P" first="P. J. M." last="Rottier">P. J. M. Rottier</name><name sortKey="Te Lintelo, E G" sort="Te Lintelo, E G" uniqKey="Te Lintelo E" first="E. G." last="Te Lintelo">E. G. Te Lintelo</name><name sortKey="Verheije, M H" sort="Verheije, M H" uniqKey="Verheije M" first="M. H." last="Verheije">M. H. Verheije</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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|area= SrasV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= Pascal:07-0517267
|texte= Localization and membrane topology of coronavirus nonstructural protein 4: Involvement of the early secretory pathway in replication
}}
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