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PMID: 23427160 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Hepatitis C virus replication is modulated by the interaction of nonstructural protein NS5B and fatty acid synthase.

Journal of virology ·Vol. 87 ·No. 9 ·2013-05-00 ·Pages 4994-5004

Huang JT, Tseng CP, Liao MH, Lu SC, Yeh WZ, Sakamoto N, Chen CM, Cheng JC

Abstract

Hepatitis C virus (HCV) nonstructural protein 5B (NS5B) is an RNA-dependent RNA polymerase (RdRp) that acts as a key player in the HCV replication complex. Understanding the interplay between the viral and cellular components of the HCV replication complex could provide new insight for prevention of the progression of HCV-associated hepatocellular carcinoma (HCC). In this study, the NS5B protein was used as the bait in a pulldown assay to screen for NS5B-interacting proteins that are present in Huh7 hepatoma cell lysates. After mass spectrophotometric analysis, fatty acid synthase (FASN) was found to interact with NS5B. Coimmunoprecipitation and double staining assays further confirmed the direct binding between NS5B and FASN. The domain of NS5B that interacts with FASN was also determined. Moreover, FASN was associated with detergent-resistant lipid rafts and colocalized with NS5B in active HCV replication complexes. In addition, overexpression of FASN enhanced HCV expression in Huh7/Rep-Feo cells, while transfection of FASN small interfering RNA (siRNA) or treatment with FASN-specific inhibitors decreased HCV replication and viral production. Notably, FASN directly increased HCV NS5B RdRp activity in vitro. These results together indicate that FASN interacts with NS5B and modulates HCV replication through a direct increase of NS5B RdRp activity. FASN may thereby serve as a target for the treatment of HCV infection and the prevention of HCV-associated HCC progression.

MeSH Terms
Cell Line, Tumor Fatty Acid Synthases/genetics,metabolism Hepacivirus/enzymology,genetics,physiology Hepatitis C/enzymology,genetics,virology Humans Protein Binding Viral Nonstructural Proteins/genetics,metabolism Virus Replication
Chemicals
NS-5 protein, hepatitis C virus Viral Nonstructural Proteins Fatty Acid Synthases
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Huang Jing-Tang
Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan.
Tseng Ching-Ping
Liao Mei-Huei
Lu Shao-Chun
Yeh Wei-Zhou
Sakamoto Naoya
Chen Chuan-Mu
Cheng Ju-Chien
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
1098-5514
DOI
10.1128/JVI.02526-12
Published
2013-05-00
Epub
2013-00-20
Pages
4994-5004
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC3624299
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.02526-12 PMC Full Text View journal details
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