Establishment and Application of a High Throughput Screening System Targeting the Interaction between HCV Internal Ribosome Entry Site and Human Eukaryotic Translation Initiation Factor 3

Yuying Zhu; Pei Huang; Na Yang; Rui Liu; Xueting Liu; Huanqin Dai; Lixin Zhang; Fuhang Song; Chaomin Sun

doi:10.3389/fmicb.2017.00977

Establishment and Application of a High Throughput Screening System Targeting the Interaction between HCV Internal Ribosome Entry Site and Human Eukaryotic Translation Initiation Factor 3

Front Microbiol. 2017 May 29:8:977. doi: 10.3389/fmicb.2017.00977. eCollection 2017.

Authors

Yuying Zhu^{1

2

3}, Pei Huang⁴, Na Yang^{1

2}, Rui Liu^{1

2}, Xueting Liu⁴, Huanqin Dai⁴, Lixin Zhang^{4

5}, Fuhang Song⁴, Chaomin Sun^{1

2}

Affiliations

¹ Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of SciencesQingdao, China.
² Laboratory for Marine Biology and Biotechnology, Qingdao National Laboratory for Marine Science and TechnologyQingdao, China.
³ College of Earth Science, University of Chinese Academy of SciencesBeijing, China.
⁴ Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of SciencesBeijing, China.
⁵ State Key Laboratory of Bioreactor Engineering, East China University of Science and TechnologyShanghai, China.

Abstract

Viruses are intracellular obligate parasites and the host cellular machinery is usually recruited for their replication. Human eukaryotic translation initiation factor 3 (eIF3) could be directly recruited by the hepatitis C virus (HCV) internal ribosome entry site (IRES) to promote the translation of viral proteins. In this study, we establish a fluorescence polarization (FP) based high throughput screening (HTS) system targeting the interaction between HCV IRES and eIF3. By screening a total of 894 compounds with this HTS system, two compounds (Mucl39526 and NP39) are found to disturb the interaction between HCV IRES and eIF3. And these two compounds are further demonstrated to inhibit the HCV IRES-dependent translation in vitro. Thus, this HTS system is functional to screen the potential HCV replication inhibitors targeting human eIF3, which is helpful to overcome the problem of viral resistance. Surprisingly, one compound HP-3, a kind of oxytocin antagonist, is discovered to significantly enhance the interaction between HCV IRES and eIF3 by this HTS system. HP-3 is demonstrated to directly interact with HCV IRES and promote the HCV IRES-dependent translation both in vitro and in vivo, which strongly suggests that HP-3 has potentials to promote HCV replication. Therefore, this HTS system is also useful to screen the potential HCV replication enhancers, which is meaningful for understanding the viral replication and screening novel antiviral drugs. To our knowledge, this is the first HTS system targeting the interaction between eIF3 and HCV IRES, which could be applied to screen both potential HCV replication inhibitors and enhancers.

Keywords: IRES; eIF3; enhancer; hepatitis C virus; high throughput; inhibitor.