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Interaction of Stomatin with Hepatitis C Virus RNA Polymerase Stabilizes the Viral RNA Replicase Complexes on Detergent-Resistant Membranes

Title
Interaction of Stomatin with Hepatitis C Virus RNA Polymerase Stabilizes the Viral RNA Replicase Complexes on Detergent-Resistant Membranes
Authors
Kim, Jung-HeeRhee, Jin-KyuAhn, Dae-GyunKim, Kwang PyoOh, Jong-Won
Ewha Authors
이진규
SCOPUS Author ID
이진규scopus
Issue Date
2014
Journal Title
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
ISSN
1017-7825JCR Link

1738-8872JCR Link
Citation
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY vol. 24, no. 12, pp. 1744 - 1754
Keywords
HCVRNA-dependent RNA polymerasestomatindetergent-resistant membraneRNA replicase complex
Publisher
KOREAN SOC MICROBIOLOGY &

BIOTECHNOLOGY
Indexed
SCIE; SCOPUS; KCI WOS
Document Type
Article
Abstract
The hepatitis C virus (HCV) RNA genome is replicated by an RNA replicase complex (RC) consisting of cellular proteins and viral nonstructural (NS) proteins, including NS5B, an RNA-dependent RNA polymerase (RdRp) and key enzyme for viral RNA genome replication. The HCV RC is known to be associated with an intracellular membrane structure, but the cellular components of the RC and their roles in the formation of the HCV RC have not been well characterized. In this study, we took a proteomic approach to identify stomatin, a member of the integral proteins of lipid rafts, as a cellular protein interacting with HCV NS5B. Co-immunoprecipitation and co-localization studies confirmed the interaction between stomatin and NS5B. We demonstrated that the subcellular fraction containing viral NS proteins and stomatin displays RdRp activity. Membrane flotation assays with the HCV genome replication-competent subcellular fraction revealed that the HCV RdRp and stomatin are associated with the lipid raft-like domain of membranous structures. Stomatin silencing by RNA interference led to the release of NS5B from the detergent-resistant membrane, thereby inhibiting HCV replication in both HCV subgenomic replicon-harboring cells and HCV-infected cells. Our results identify stomatin as a cellular protein that plays a role in the formation of an enzymatically active HCV RC on a detergent-resistant membrane structure.
DOI
10.4014/jmb.1409.09063
Appears in Collections:
엘텍공과대학 > 식품공학전공 > Journal papers
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