The transmembrane domain of the severe acute respiratory syndrome coronavirus ORF7b protein is necessary and sufficient for its retention in the Golgi complex.
Identifieur interne : 003050 ( Main/Merge ); précédent : 003049; suivant : 003051The transmembrane domain of the severe acute respiratory syndrome coronavirus ORF7b protein is necessary and sufficient for its retention in the Golgi complex.
Auteurs : Scott R. Schaecher [États-Unis] ; Michael S. Diamond ; Andrew PekoszSource :
- Journal of virology [ 1098-5514 ] ; 2008.
Descripteurs français
- KwdFr :
- Animaux, Appareil de Golgi (métabolisme), Cellules Vero, Données de séquences moléculaires, Humains, Membrane cellulaire (virologie), Modèles biologiques, Protéines virales (métabolisme), Similitude de séquences d'acides aminés, Structure tertiaire des protéines, Séquence d'acides aminés, Technique d'immunofluorescence indirecte, Virus du SRAS (génétique).
- MESH :
- génétique : Virus du SRAS.
- métabolisme : Appareil de Golgi, Protéines virales.
- virologie : Membrane cellulaire.
- Animaux, Cellules Vero, Données de séquences moléculaires, Humains, Modèles biologiques, Similitude de séquences d'acides aminés, Structure tertiaire des protéines, Séquence d'acides aminés, Technique d'immunofluorescence indirecte.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, CD4 Antigens (chemistry), Cell Membrane (virology), Chlorocebus aethiops, Fluorescent Antibody Technique, Indirect, Golgi Apparatus (metabolism), Humans, Models, Biological, Molecular Sequence Data, Protein Structure, Tertiary, SARS Virus (genetics), Sequence Homology, Amino Acid, Vero Cells, Viral Proteins (metabolism).
- MESH :
- chemical , chemistry : CD4 Antigens.
- genetics : SARS Virus.
- metabolism : Golgi Apparatus, Viral Proteins.
- virology : Cell Membrane.
- Amino Acid Sequence, Animals, Chlorocebus aethiops, Fluorescent Antibody Technique, Indirect, Humans, Models, Biological, Molecular Sequence Data, Protein Structure, Tertiary, Sequence Homology, Amino Acid, Vero Cells.
Abstract
The severe acute respiratory syndrome coronavirus (SARS-CoV) ORF7b (also called 7b) protein is an integral membrane protein that is translated from a bicistronic open reading frame encoded within subgenomic RNA 7. When expressed independently or during virus infection, ORF7b accumulates in the Golgi compartment, colocalizing with both cis- and trans-Golgi markers. To identify the domains of this protein that are responsible for Golgi localization, we have generated a set of mutant proteins and analyzed their subcellular localizations by indirect immunofluorescence confocal microscopy. The N- and C-terminal sequences are dispensable, but the ORF7b transmembrane domain (TMD) is essential for Golgi compartment localization. When the TMD of human CD4 was replaced with the ORF7b TMD, the resulting chimeric protein localized to the Golgi complex. Scanning alanine mutagenesis identified two regions in the carboxy-terminal portion of the TMD that eliminated the Golgi complex localization of the chimeric CD4 proteins or ORF7b protein. Collectively, these data demonstrate that the Golgi complex retention signal of the ORF7b protein resides solely within the TMD.
DOI: 10.1128/JVI.00784-08
PubMed: 18632859
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<term>Technique d'immunofluorescence indirecte</term>
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<term>Modèles biologiques</term>
<term>Similitude de séquences d'acides aminés</term>
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<front><div type="abstract" xml:lang="en">The severe acute respiratory syndrome coronavirus (SARS-CoV) ORF7b (also called 7b) protein is an integral membrane protein that is translated from a bicistronic open reading frame encoded within subgenomic RNA 7. When expressed independently or during virus infection, ORF7b accumulates in the Golgi compartment, colocalizing with both cis- and trans-Golgi markers. To identify the domains of this protein that are responsible for Golgi localization, we have generated a set of mutant proteins and analyzed their subcellular localizations by indirect immunofluorescence confocal microscopy. The N- and C-terminal sequences are dispensable, but the ORF7b transmembrane domain (TMD) is essential for Golgi compartment localization. When the TMD of human CD4 was replaced with the ORF7b TMD, the resulting chimeric protein localized to the Golgi complex. Scanning alanine mutagenesis identified two regions in the carboxy-terminal portion of the TMD that eliminated the Golgi complex localization of the chimeric CD4 proteins or ORF7b protein. Collectively, these data demonstrate that the Golgi complex retention signal of the ORF7b protein resides solely within the TMD.</div>
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