Membrane association of the RNA-dependent RNA polymerase is essential for hepatitis C virus RNA replication.

Details

Serval ID
serval:BIB_5AD3E4240A81
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Membrane association of the RNA-dependent RNA polymerase is essential for hepatitis C virus RNA replication.
Journal
Journal of virology
Author(s)
Moradpour D., Brass V., Bieck E., Friebe P., Gosert R., Blum H.E., Bartenschlager R., Penin F., Lohmann V.
ISSN
0022-538X
Publication state
Published
Issued date
2004
Peer-reviewed
Oui
Volume
78
Number
23
Pages
13278-84
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Abstract
The hepatitis C virus (HCV) RNA-dependent RNA polymerase (RdRp), represented by nonstructural protein 5B (NS5B), belongs to a class of integral membrane proteins termed tail-anchored proteins. Its membrane association is mediated by the C-terminal 21 amino acid residues, which are dispensable for RdRp activity in vitro. For this study, we investigated the role of this domain, termed the insertion sequence, in HCV RNA replication in cells. Based on a structural model and the amino acid conservation among different HCV isolates, we designed a panel of insertion sequence mutants and analyzed their membrane association and RNA replication. Subgenomic replicons with a duplication of an essential cis-acting replication element overlapping the sequence that encodes the C-terminal domain of NS5B were used to unequivocally distinguish RNA versus protein effects of these mutations. Our results demonstrate that the membrane association of the RdRp is essential for HCV RNA replication. Interestingly, certain amino acid substitutions within the insertion sequence abolished RNA replication without affecting membrane association, indicating that the C-terminal domain of NS5B has functions beyond serving as a membrane anchor and that it may be involved in critical intramembrane protein-protein interactions. These results have implications for the functional architecture of the HCV replication complex and provide new insights into the expanding spectrum of tail-anchored proteins.
Keywords
Amino Acid Sequence, Base Sequence, Hepacivirus, RNA Replicase, RNA, Viral, Viral Nonstructural Proteins, Virus Replication
Pubmed
Web of science
Open Access
Yes
Create date
25/01/2008 17:05
Last modification date
20/08/2019 15:13
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