Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines

Détails

ID Serval
serval:BIB_FC14AB8D86F8
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3-NS4A complex expressed in tetracycline-regulated cell lines
Périodique
Journal of Virology
Auteur⸱e⸱s
Wolk  B., Sansonno  D., Krausslich  H. G., Dammacco  F., Rice  C. M., Blum  H. E., Moradpour  D.
ISSN
0022-538X (Print)
Statut éditorial
Publié
Date de publication
03/2000
Volume
74
Numéro
5
Pages
2293-304
Notes
Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S. --- Old month value: Mar
Résumé
A tetracycline-regulated gene expression system and a panel of novel monoclonal antibodies were used to examine the subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus (HCV) NS3-NS4A complex in inducible cell lines. The NS3 serine protease domain and the full-length NS3 protein expressed in the absence of the NS4A cofactor were diffusely distributed in the cytoplasm and nucleus. Coexpression of NS4A, however, directed NS3 to the endoplasmic reticulum (ER) or an ER-like modified compartment, as demonstrated by colocalization with 3,3'-dihexyloxacarbocyanine iodide, protein disulfide isomerase, and calnexin, as well as subcellular fractionation analyses. In addition, coexpression with NS4A dramatically increased the intracellular stability of NS3 (mean protein half-life of 26 versus 3 h) and allowed for NS4A-dependent trans-cleavage at the NS4B-NS5A junction. Deletion analyses revealed that the hydrophobic amino-terminal domain of NS4A was required for ER targeting of NS3. These results demonstrate the importance of studying HCV proteins in their biological context and define a well-characterized cell culture system for further analyses of the NS3-NS4A complex and the evaluation of novel antiviral strategies against hepatitis C.
Mots-clé
Animals Antibodies, Monoclonal/immunology Cell Line Cells, Cultured Culture Media Cytoplasm/virology Endoplasmic Reticulum/virology Fluorescent Antibody Technique Hepacivirus/metabolism Hepatitis C, Chronic/metabolism Humans Immunoblotting Liver/virology Macromolecular Substances Mice Mice, Inbred BALB C Microscopy, Confocal Subcellular Fractions/virology Tetracycline Transformation, Genetic Viral Nonstructural Proteins/*analysis/genetics/immunology
Pubmed
Web of science
Création de la notice
25/01/2008 16:05
Dernière modification de la notice
20/08/2019 16:27
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