HEV-ORF3 Encoding Phosphoprotein Interacts With Hepsin |
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| Authors: | Chunyan Wang Liang Guo Dayi Yu Xiuguo Hua Zhibiao Yang Congli Yuan Li Cui |
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| Institution: | 1.Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China;2.Animal Disease Control Center of Min Hang District, Shanghai, China |
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| Abstract: | Background:Hepatitis E virus (HEV) is a major causative agent of acute clinical hepatitis in adults through much of Asia, the Middle East and Africa. Open reading frame 3 (ORF3) encodes around 120 amino acids of phosphorylation protein that associates with the cytoskeleton, while its precise biological function is still unknown.Objectives:In order to understand the function of ORF3 protein (pORF3) in depth, HEV ORF3 interacting proteins were screened in human hepatocytes cDNA library using two-hybrid system techniques and further verification of the interactions were carried out through co-immunoprecipitation (Co-IP).Materials and Methods:The Cyto-Trap two-hybrid system technology, a classical method for analyzing protein interactions, was used to screen the pORF3 interacting proteins from human hepatocytes cDNA library.Results:Through the Cyto-Trap two-hybrid system, eight proteins interacting with pORF3 were winnowed. The Co-IP results confirmed that hepsin which is reported to function as the inhibitor of several tumors reacted with pORF3.Conclusions:Out of eight screened proteins interacting with pORF3, hepsin was confirmed to have specific interactions with pORF3. |
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| Keywords: | ORF3 protein Hepatitis E virus Hepsin Two-Hybrid System Techniques Immunoprecipitation |
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