Mapping the Membrane Topology of Hepatitis C Virus Nonstructural Protein 4B

Details

Serval ID
serval:BIB_78214751B3F7
Type
Inproceedings: an article in a conference proceedings.
Publication sub-type
Abstract (Abstract): shot summary in a article that contain essentials elements presented during a scientific conference, lecture or from a poster.
Collection
Publications
Title
Mapping the Membrane Topology of Hepatitis C Virus Nonstructural Protein 4B
Title of the conference
Annual Meeting of the Swiss Society of Gastroenterology, Swiss Society for Visceral, Surgery Swiss Association for the Study of the Liver, Swiss Association of Clinical Nutrition
Author(s)
Gouttenoire J., Roulin P., Penin F., Moradpour D.
Address
Interlaken, Switzerland, September 23-24, 2010
ISSN
1424-7860
Publication state
Published
Issued date
2010
Peer-reviewed
Oui
Volume
140
Series
Swiss Medical Weekly
Pages
6S
Language
english
Notes
Meeting Abstract
Abstract
BACKGROUND: Nonstructural protein 4B (NS4B) plays an essential role in the formation of the hepatitis C virus (HCV) replication complex. It is an integral membrane protein that has only poorly been characterized to date. In particular, a precise membrane topology is thus far elusive. Here, we explored a novel strategy to map the membrane topology of HCV NS4B.
METHODS: Selective permeabilization of the plasma membrane, maleimide-polyethyleneglycol (mPEG) labeling of natural or engineered cysteine residues and immunoblot analyses were combined to map the membrane topology of NS4B. Cysteine substitutions were introduced at carefully selected positions within NS4B and their impact on HCV RNA replication and infectious virus production analyzed in cell culture.
RESULTS: We established a panel of viable HCV mutants with cysteine substitutions at strategic positions within NS4B. These mutants are infectious and replicate to high levels in cell culture. In parallel, we adapted and optimized the selective permeabilization and mPEG labeling techniques to Huh-7 human hepatocellular carcinoma cells which can support HCV infection and replication.
CONCLUSIONS: The newly established experimental tools and techniques should allow us to refine the membrane topology of HCV NS4B in a physiological context. The expected results should enhance our understanding of the functional architecture of the HCV replication complex and may provide new opportunities for antiviral intervention in the future.
Web of science
Create date
01/10/2015 15:16
Last modification date
21/08/2019 6:34
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