Inhibition of hepatitis C virus infection by DNA aptamer against NS2 protein

Yimin Gao; Xiaoyan Yu; Binbin Xue; Fei Zhou; Xiaohong Wang; Darong Yang; Nianli Liu; Li Xu; Xiaohong Fang; Haizhen Zhu

doi:10.1371/journal.pone.0090333

Inhibition of hepatitis C virus infection by DNA aptamer against NS2 protein

PLoS One. 2014 Feb 28;9(2):e90333. doi: 10.1371/journal.pone.0090333. eCollection 2014.

Authors

Yimin Gao¹, Xiaoyan Yu¹, Binbin Xue¹, Fei Zhou¹, Xiaohong Wang¹, Darong Yang², Nianli Liu², Li Xu³, Xiaohong Fang³, Haizhen Zhu²

Affiliations

¹ Department of Molecular Medicine, State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, China.
² Department of Molecular Medicine, State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha, China ; Research Center of Cancer Prevention & Treatment and Translational Medicine Research Center of Liver Cancer, Hunan Provincial Tumor Hospital (Affiliated Tumor Hospital of Xiangya Medical School of Central South University), Changsha, China.
³ Key Laboratory of Molecular Nanostructure and Nanotechnology, Institute of Chemistry, Chinese Academy of Sciences, Beijing, China.

Abstract

NS2 protein is essential for hepatitis C virus (HCV) replication. NS2 protein was expressed and purified. Aptamers against NS2 protein were raised and antiviral effects of the aptamers were examined. The molecular mechanism through which the aptamers exert their anti-HCV activity was investigated. The data showed that aptamer NS2-3 inhibited HCV RNA replication in replicon cell line and infectious HCV cell culture system. NS2-3 and another aptamer NS2-2 were demonstrated to inhibit infectious virus production without cytotoxicity in vitro. They did not affect hepatitis B virus replication. Interferon beta (IFN-β) and interferon-stimulated genes (ISGs) were not induced by the aptamers in HCV-infected hepatocytes. Furthermore, our study showed that N-terminal region of NS2 protein is involved in the inhibition of HCV infection by NS2-2. I861T within NS2 is the major resistance mutation identified. Aptamer NS2-2 disrupts the interaction of NS2 with NS5A protein. The data suggest that NS2-2 aptamer against NS2 protein exerts its antiviral effects through binding to the N-terminal of NS2 and disrupting the interaction of NS2 with NS5A protein. NS2-specific aptamer is the first NS2 inhibitor and can be used to understand the mechanisms of virus replication and assembly. It may be served as attractive candidates for inclusion in the future HCV direct-acting antiviral combination therapies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antiviral Agents / pharmacology*
Aptamers, Nucleotide / biosynthesis
Aptamers, Nucleotide / genetics
Aptamers, Nucleotide / pharmacology*
Base Sequence
Binding Sites
Cell Line, Tumor
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression Regulation, Viral*
Hepacivirus / drug effects*
Hepacivirus / physiology
Hepatocytes / drug effects*
Hepatocytes / immunology
Hepatocytes / virology
Host-Pathogen Interactions
Humans
Interferon-beta
Molecular Sequence Data
Protein Binding
Recombinant Proteins / biosynthesis
Recombinant Proteins / genetics
Recombinant Proteins / pharmacology
Viral Nonstructural Proteins / antagonists & inhibitors
Viral Nonstructural Proteins / genetics*
Viral Nonstructural Proteins / metabolism
Virus Replication

Substances

Antiviral Agents
Aptamers, Nucleotide
NS2 protein, Hepatitis C virus
Recombinant Proteins
Viral Nonstructural Proteins
Interferon-beta
NS-5 protein, hepatitis C virus

Grants and funding

This work was supported by National Science and Technology Major Project of the Ministry of Science and Technology of China (2009ZX10004-312), National Natural Science Foundation of China (81271885), and Program for New Century Excellent Talents in University (NCET-09-0339) to HZ. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.