SrasV1, Ncbi, Merge, bibRecord, 000A28

Identification and characterization of severe acute respiratory syndrome coronavirus replicase proteins.

Identifieur interne : 000A28 ( Ncbi/Merge ); précédent : 000A27; suivant : 000A29

Identification and characterization of severe acute respiratory syndrome coronavirus replicase proteins.

Auteurs : Erik Prentice [États-Unis] ; Josephine Mcauliffe ; Xiaotao Lu ; Kanta Subbarao ; Mark R. Denison

Source :

Journal of virology [ 0022-538X ] ; 2004.

RBID : pubmed:15331731

Descripteurs français

KwdFr :
- ADN viral (génétique), Animaux, Anticorps antiviraux, Cadres ouverts de lecture, Cellules Vero, Immunotransfert, Protéines virales (génétique), Protéines virales (immunologie), Protéines virales (isolement et purification), Protéines virales (métabolisme), RNA replicase (génétique), RNA replicase (immunologie), RNA replicase (isolement et purification), RNA replicase (métabolisme), Séquence nucléotidique, Technique d'immunofluorescence, Virus de l'hépatite murine (enzymologie), Virus de l'hépatite murine (génétique), Virus du SRAS (enzymologie), Virus du SRAS (génétique), Virus du SRAS (pathogénicité), Épitopes (génétique).
MESH :
- enzymologie : Virus de l'hépatite murine, Virus du SRAS.
- génétique : ADN viral, Protéines virales, RNA replicase, Virus de l'hépatite murine, Virus du SRAS, Épitopes.
- immunologie : Protéines virales, RNA replicase.
- isolement et purification : Protéines virales, RNA replicase.
- métabolisme : Protéines virales, RNA replicase.
- pathogénicité : Virus du SRAS.
- Animaux, Anticorps antiviraux, Cadres ouverts de lecture, Cellules Vero, Immunotransfert, Séquence nucléotidique, Technique d'immunofluorescence.

English descriptors

KwdEn :
- Animals, Antibodies, Viral, Base Sequence, Chlorocebus aethiops, DNA, Viral (genetics), Epitopes (genetics), Fluorescent Antibody Technique, Immunoblotting, Murine hepatitis virus (enzymology), Murine hepatitis virus (genetics), Open Reading Frames, RNA Replicase (genetics), RNA Replicase (immunology), RNA Replicase (isolation & purification), RNA Replicase (metabolism), SARS Virus (enzymology), SARS Virus (genetics), SARS Virus (pathogenicity), Vero Cells, Viral Proteins (genetics), Viral Proteins (immunology), Viral Proteins (isolation & purification), Viral Proteins (metabolism).
MESH :
- chemical , genetics : DNA, Viral, Epitopes, RNA Replicase, Viral Proteins.
- chemical , immunology : RNA Replicase, Viral Proteins.
- chemical , isolation & purification : RNA Replicase, Viral Proteins.
- chemical , metabolism : RNA Replicase, Viral Proteins.
- chemical : Antibodies, Viral.
- enzymology : Murine hepatitis virus, SARS Virus.
- genetics : Murine hepatitis virus, SARS Virus.
- pathogenicity : SARS Virus.
- Animals, Base Sequence, Chlorocebus aethiops, Fluorescent Antibody Technique, Immunoblotting, Open Reading Frames, Vero Cells.

Abstract

The severe acute respiratory syndrome coronavirus (SARS-CoV) encodes proteins required for RNA transcription and genome replication as large polyproteins that are proteolytically processed by virus-encoded proteinases to produce mature replicase proteins. In this report, we generated antibodies against SARS-CoV predicted replicase protein and used the antibodies to identify and characterize 12 of the 16 predicted mature replicase proteins (nsp1, nsp2, nsp3, nsp4, nsp5, nsp8, nsp9, nsp12, nsp13, nsp14, nsp15, and nsp16) in SARS-CoV-infected Vero cells. Immunoblot analysis of infected-cell lysates identified proteins of the predicted sizes. Immunofluorescence microscopy detected similar patterns of punctate perinuclear and distributed cytoplasmic foci with all replicase antibodies and as early as 6 h postinfection. Dual-labeling studies demonstrated colocalization of replicase protein nsp8 with nsp2 and nsp3 in cytoplasmic complexes and also with LC3, a protein marker for autophagic vacuoles. Antibodies directed against mouse hepatitis virus (MHV) virions and against the putative RNA-dependent RNA polymerase (Pol) detected SARS-CoV nucleocapsid and nsp12 (Pol), respectively, in SARS-CoV-infected Vero cells. These results confirm the predicted protein processing pattern for mature SARS-CoV replicase proteins, demonstrate localization of replicase proteins to cytoplasmic complexes containing markers for autophagosome membranes, and suggest conservation of protein epitopes in the replicase and nucleocapsid of SARS-CoV and the group II coronavirus, MHV. Further, the results demonstrate the ability of replicase antibodies to detect SARS-CoV-infected cells as early as 6 h postinfection and thus represent important tools for studies of SARS-CoV replication, inhibition, and diagnosis.

DOI: 10.1128/JVI.78.18.9977-9986.2004
PubMed: 15331731

Links toward previous steps (curation, corpus...)

to stream PubMed, to step Corpus: 002B96
to stream PubMed, to step Curation: 002B96
to stream PubMed, to step Checkpoint: 002C67

Links to Exploration step

pubmed:15331731

Le document en format XML

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<front><div type="abstract" xml:lang="en">The severe acute respiratory syndrome coronavirus (SARS-CoV) encodes proteins required for RNA transcription and genome replication as large polyproteins that are proteolytically processed by virus-encoded proteinases to produce mature replicase proteins. In this report, we generated antibodies against SARS-CoV predicted replicase protein and used the antibodies to identify and characterize 12 of the 16 predicted mature replicase proteins (nsp1, nsp2, nsp3, nsp4, nsp5, nsp8, nsp9, nsp12, nsp13, nsp14, nsp15, and nsp16) in SARS-CoV-infected Vero cells. Immunoblot analysis of infected-cell lysates identified proteins of the predicted sizes. Immunofluorescence microscopy detected similar patterns of punctate perinuclear and distributed cytoplasmic foci with all replicase antibodies and as early as 6 h postinfection. Dual-labeling studies demonstrated colocalization of replicase protein nsp8 with nsp2 and nsp3 in cytoplasmic complexes and also with LC3, a protein marker for autophagic vacuoles. Antibodies directed against mouse hepatitis virus (MHV) virions and against the putative RNA-dependent RNA polymerase (Pol) detected SARS-CoV nucleocapsid and nsp12 (Pol), respectively, in SARS-CoV-infected Vero cells. These results confirm the predicted protein processing pattern for mature SARS-CoV replicase proteins, demonstrate localization of replicase proteins to cytoplasmic complexes containing markers for autophagosome membranes, and suggest conservation of protein epitopes in the replicase and nucleocapsid of SARS-CoV and the group II coronavirus, MHV. Further, the results demonstrate the ability of replicase antibodies to detect SARS-CoV-infected cells as early as 6 h postinfection and thus represent important tools for studies of SARS-CoV replication, inhibition, and diagnosis.</div>
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<Abstract><AbstractText>The severe acute respiratory syndrome coronavirus (SARS-CoV) encodes proteins required for RNA transcription and genome replication as large polyproteins that are proteolytically processed by virus-encoded proteinases to produce mature replicase proteins. In this report, we generated antibodies against SARS-CoV predicted replicase protein and used the antibodies to identify and characterize 12 of the 16 predicted mature replicase proteins (nsp1, nsp2, nsp3, nsp4, nsp5, nsp8, nsp9, nsp12, nsp13, nsp14, nsp15, and nsp16) in SARS-CoV-infected Vero cells. Immunoblot analysis of infected-cell lysates identified proteins of the predicted sizes. Immunofluorescence microscopy detected similar patterns of punctate perinuclear and distributed cytoplasmic foci with all replicase antibodies and as early as 6 h postinfection. Dual-labeling studies demonstrated colocalization of replicase protein nsp8 with nsp2 and nsp3 in cytoplasmic complexes and also with LC3, a protein marker for autophagic vacuoles. Antibodies directed against mouse hepatitis virus (MHV) virions and against the putative RNA-dependent RNA polymerase (Pol) detected SARS-CoV nucleocapsid and nsp12 (Pol), respectively, in SARS-CoV-infected Vero cells. These results confirm the predicted protein processing pattern for mature SARS-CoV replicase proteins, demonstrate localization of replicase proteins to cytoplasmic complexes containing markers for autophagosome membranes, and suggest conservation of protein epitopes in the replicase and nucleocapsid of SARS-CoV and the group II coronavirus, MHV. Further, the results demonstrate the ability of replicase antibodies to detect SARS-CoV-infected cells as early as 6 h postinfection and thus represent important tools for studies of SARS-CoV replication, inhibition, and diagnosis.</AbstractText>
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Identification and characterization of severe acute respiratory syndrome coronavirus replicase proteins.

Identification and characterization of severe acute respiratory syndrome coronavirus replicase proteins.

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