Abstract
Hepatitis C virus (HCV) nonstructural protein 5A (NS5A) is a component of the replication complex associated with various cellular proteins. It has been reported that G protein pathway suppressor 2 (GPS2) is a potential NS5A-binding factor, as identified in a yeast two-hybrid screens of human cDNA library using viral proteins as baits [1]. In this study, we demonstrated the interaction between GPS2 and NS5A in mammalian cells by coimmunoprecipitation analysis and found that both exogenously and endogenously expressed GPS2 interacted with NS5A of genotype 1b and 2a. Mutagenesis study demonstrated that Domain I of NS5A and coiled-coil domain of GPS2 are responsible for the interaction. Knockdown of GPS2 in hepatoma cell lines suppressed the replication of HCV RNA, which can be rescued by the expression of an RNAi-resistant GPS2. Furthermore, overexpression of GPS2 enhanced the association of NS5A with a proviral cellular factor, human vesicle-associated membrane protein-associated protein A (VAP-A), while knockdown of GPS2 disrupted interaction between VAP-A and NS5A. Taken together, our results suggest that GPS2 acts as a bridge between NS5A and VAP-A and is required for efficient HCV replication.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Binding Sites
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Cell Line, Tumor
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Gene Expression Regulation
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Hepacivirus / genetics
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Hepacivirus / metabolism*
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Hepatocytes / virology*
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Host-Pathogen Interactions
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Humans
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Immunoprecipitation
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Intracellular Signaling Peptides and Proteins / antagonists & inhibitors
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Intracellular Signaling Peptides and Proteins / genetics
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Intracellular Signaling Peptides and Proteins / metabolism*
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Protein Binding
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Protein Interaction Domains and Motifs
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Signal Transduction
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Vesicular Transport Proteins / genetics
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Vesicular Transport Proteins / metabolism*
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Viral Nonstructural Proteins / genetics
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Viral Nonstructural Proteins / metabolism*
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Virus Replication
Substances
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GPS2 protein, human
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Intracellular Signaling Peptides and Proteins
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RNA, Small Interfering
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VAPA protein, human
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Vesicular Transport Proteins
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Viral Nonstructural Proteins
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NS-5 protein, hepatitis C virus
Grants and funding
This work was supported by the National Basic Research Program of China (Grant No. 2011CB504800). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.