Interaction of Stomatin with Hepatitis C virus RNA Polymerase stabilizes the viral RNA Replicase complexes on detergent-resistant membranes

Jung Hee Kim, Jin Kyu Rhee, Dae Gyun Ahn, Kwang Pyol Kim, Jong Won Oh

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 RNAdependent 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. Coimmunoprecipitation 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 HCVinfected 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.

Original languageEnglish
Pages (from-to)1744-1754
Number of pages11
JournalJournal of microbiology and biotechnology
Volume24
Issue number12
DOIs
Publication statusPublished - 2014 Jan 1

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RNA Replicase
Viral RNA
DNA-Directed RNA Polymerases
Hepacivirus
Detergents
Membranes
Viral Nonstructural Proteins
Subcellular Fractions
Virus Replication
Proteins
Genome
Lipids
Intracellular Membranes
Replicon
Viral Genome
Cellular Structures
RNA Interference
Proteomics
RNA

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

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title = "Interaction of Stomatin with Hepatitis C virus RNA Polymerase stabilizes the viral RNA Replicase complexes on detergent-resistant membranes",
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 RNAdependent 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. Coimmunoprecipitation 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 HCVinfected 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.",
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Interaction of Stomatin with Hepatitis C virus RNA Polymerase stabilizes the viral RNA Replicase complexes on detergent-resistant membranes. / Kim, Jung Hee; Rhee, Jin Kyu; Ahn, Dae Gyun; Kim, Kwang Pyol; Oh, Jong Won.

In: Journal of microbiology and biotechnology, Vol. 24, No. 12, 01.01.2014, p. 1744-1754.

Research output: Contribution to journalArticle

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