SrasV1, Main, Exploration, bibRecord, 004404

Ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex

Identifieur interne : 004404 ( Main/Exploration ); précédent : 004403; suivant : 004405

Ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex

Auteurs : Eric J. Snijder [Pays-Bas] ; Yvonne Van Der Meer [Pays-Bas] ; Jessika Zevenhoven-Dobbe [Pays-Bas] ; Jos J. M. Ondervater [Pays-Bas] ; Jannes Van Der Meulen [Pays-Bas] ; Henk K. Koerten [Pays-Bas] ; A. Mieke Mommaas [Pays-Bas]

Source :

Journal of virology [ 0022-538X ] ; 2006.

RBID : Pascal:06-0310720

Descripteurs français

KwdFr :
- Animaux, Cellules Vero, Membranes intracellulaires (métabolisme), Membranes intracellulaires (ultrastructure), Microscopie électronique, Origine de réplication, RNA replicase (analyse), Réplication virale, Réticulum endoplasmique (microbiologie), Réticulum endoplasmique (virologie), Virus du SRAS, Vésicules de transport (ultrastructure).
MESH :
- analyse : RNA replicase.
- microbiologie : Réticulum endoplasmique.
- métabolisme : Membranes intracellulaires.
- virologie : Animaux, Cellules Vero, Membranes intracellulaires, Microscopie électronique, Origine de réplication, Réplication virale, Réticulum endoplasmique, Virus du SRAS, Vésicules de transport.
Pascal (Inist)
- Coronavirus, Ultrastructure, Origine, Réplication, Microbiologie, Virologie, Syndrome respiratoire aigu sévère.

English descriptors

KwdEn :
- Animals, Chlorocebus aethiops (physiology), Coronavirus, Endoplasmic Reticulum (microbiology), Endoplasmic Reticulum (virology), Intracellular Membranes (metabolism), Intracellular Membranes (ultrastructure), Microbiology, Microscopy, Electron, Origin, RNA Replicase (analysis), Replication, Replication Origin, SARS Virus, Severe acute respiratory syndrome, Transport Vesicles (ultrastructure), Ultrastructure, Vero Cells, Virology, Virus Replication.
MESH :
- chemical , analysis : RNA Replicase.
- metabolism : Intracellular Membranes.
- microbiology : Endoplasmic Reticulum.
- physiology : Chlorocebus aethiops.
- ultrastructure : Intracellular Membranes, Transport Vesicles.
- virology : Endoplasmic Reticulum.
- Animals, Microscopy, Electron, Replication Origin, SARS Virus, Vero Cells, Virus Replication.

Abstract

The RNA replication complexes of mammalian positive-stranded RNA viruses are generally associated with (modified) intracellular membranes, a feature thought to be important for creating an environment suitable for viral RNA synthesis, recruitment of host components, and possibly evasion of host defense mechanisms. Here, using a panel ofreplicase-specific antisera, we have analyzed the earlier stages of severe acute respiratory syndrome coronavirus (SARS-CoV) infection in Vero E6 cells, in particular focusing on the subcellular localization of the replicase and the ultrastructure of the associated membranes. Confocal immunofluorescence microscopy demonstrated the colocalization, throughout infection, of replicase cleavage products containing different key enzymes for SARS-CoV replication. Electron microscopy revealed the early formation and accumulation of typical double-membrane vesicles, which probably carry the viral replication complex. The vesicles appear to be fragile, and their preservation was significantly improved by using cryofixation protocols and freeze substitution methods. In immunoelectron microscopy, the virus-induced vesicles could be labeled with replicase-specific antibodies. Opposite to what was described for mouse hepatitis virus, we did not observe the late relocalization of specific replicase subunits to the presumed site of virus assembly, which was labeled using an antiserum against the viral membrane protein. This conclusion was further supported using organelle-specific marker proteins and electron microscopy. Similar morphological studies and labeling experiments argued against the previously proposed involvement of the autophagic pathway as the source for the vesicles with which the replicase is associated and instead suggested the endoplasmic reticulum to be the most likely donor of the membranes that carry the SARS-CoV replication complex.

Affiliations:

Pays-Bas

Le document en format XML

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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Chlorocebus aethiops (physiology)</term>
<term>Coronavirus</term>
<term>Endoplasmic Reticulum (microbiology)</term>
<term>Endoplasmic Reticulum (virology)</term>
<term>Intracellular Membranes (metabolism)</term>
<term>Intracellular Membranes (ultrastructure)</term>
<term>Microbiology</term>
<term>Microscopy, Electron</term>
<term>Origin</term>
<term>RNA Replicase (analysis)</term>
<term>Replication</term>
<term>Replication Origin</term>
<term>SARS Virus</term>
<term>Severe acute respiratory syndrome</term>
<term>Transport Vesicles (ultrastructure)</term>
<term>Ultrastructure</term>
<term>Vero Cells</term>
<term>Virology</term>
<term>Virus Replication</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Cellules Vero</term>
<term>Membranes intracellulaires (métabolisme)</term>
<term>Membranes intracellulaires (ultrastructure)</term>
<term>Microscopie électronique</term>
<term>Origine de réplication</term>
<term>RNA replicase (analyse)</term>
<term>Réplication virale</term>
<term>Réticulum endoplasmique (microbiologie)</term>
<term>Réticulum endoplasmique (virologie)</term>
<term>Virus du SRAS</term>
<term>Vésicules de transport (ultrastructure)</term>
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<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>RNA Replicase</term>
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<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>RNA replicase</term>
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<keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Animaux</term>
<term>Cellules Vero</term>
<term>Membranes intracellulaires</term>
<term>Microscopie électronique</term>
<term>Origine de réplication</term>
<term>Réplication virale</term>
<term>Réticulum endoplasmique</term>
<term>Virus du SRAS</term>
<term>Vésicules de transport</term>
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<keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Endoplasmic Reticulum</term>
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<term>Microscopy, Electron</term>
<term>Replication Origin</term>
<term>SARS Virus</term>
<term>Vero Cells</term>
<term>Virus Replication</term>
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<keywords scheme="Pascal" xml:lang="fr"><term>Coronavirus</term>
<term>Ultrastructure</term>
<term>Origine</term>
<term>Réplication</term>
<term>Microbiologie</term>
<term>Virologie</term>
<term>Syndrome respiratoire aigu sévère</term>
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<front><div type="abstract" xml:lang="en">The RNA replication complexes of mammalian positive-stranded RNA viruses are generally associated with (modified) intracellular membranes, a feature thought to be important for creating an environment suitable for viral RNA synthesis, recruitment of host components, and possibly evasion of host defense mechanisms. Here, using a panel ofreplicase-specific antisera, we have analyzed the earlier stages of severe acute respiratory syndrome coronavirus (SARS-CoV) infection in Vero E6 cells, in particular focusing on the subcellular localization of the replicase and the ultrastructure of the associated membranes. Confocal immunofluorescence microscopy demonstrated the colocalization, throughout infection, of replicase cleavage products containing different key enzymes for SARS-CoV replication. Electron microscopy revealed the early formation and accumulation of typical double-membrane vesicles, which probably carry the viral replication complex. The vesicles appear to be fragile, and their preservation was significantly improved by using cryofixation protocols and freeze substitution methods. In immunoelectron microscopy, the virus-induced vesicles could be labeled with replicase-specific antibodies. Opposite to what was described for mouse hepatitis virus, we did not observe the late relocalization of specific replicase subunits to the presumed site of virus assembly, which was labeled using an antiserum against the viral membrane protein. This conclusion was further supported using organelle-specific marker proteins and electron microscopy. Similar morphological studies and labeling experiments argued against the previously proposed involvement of the autophagic pathway as the source for the vesicles with which the replicase is associated and instead suggested the endoplasmic reticulum to be the most likely donor of the membranes that carry the SARS-CoV replication complex.</div>
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<tree><country name="Pays-Bas"><noRegion><name sortKey="Snijder, Eric J" sort="Snijder, Eric J" uniqKey="Snijder E" first="Eric J." last="Snijder">Eric J. Snijder</name>
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<name sortKey="Koerten, Henk K" sort="Koerten, Henk K" uniqKey="Koerten H" first="Henk K." last="Koerten">Henk K. Koerten</name>
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<name sortKey="Van Der Meulen, Jannes" sort="Van Der Meulen, Jannes" uniqKey="Van Der Meulen J" first="Jannes" last="Van Der Meulen">Jannes Van Der Meulen</name>
<name sortKey="Zevenhoven Dobbe, Jessika" sort="Zevenhoven Dobbe, Jessika" uniqKey="Zevenhoven Dobbe J" first="Jessika" last="Zevenhoven-Dobbe">Jessika Zevenhoven-Dobbe</name>
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Ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex

Ultrastructure and origin of membrane vesicles associated with the severe acute respiratory syndrome coronavirus replication complex

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Descripteurs français

English descriptors

Abstract

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