title: HEV-ORF3 Encoding Phosphoprotein Interacts With Hepsin.
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.
  
  
  OBJECTIVE
  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).
  
  
  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.
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Chunyan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Guo
authors.affiliation: Animal Disease Control Center of Min Hang District, Shanghai, China.
authors.forename: Dayi
authors.initials: D
authors.lastname: Yu
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Xiuguo
authors.initials: X
authors.lastname: Hua
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Zhibiao
authors.initials: Z
authors.lastname: Yang
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Congli
authors.initials: C
authors.lastname: Yuan
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Li
authors.initials: L
authors.lastname: Cui
keywords: Hepsin; Immunoprecipitation; ORF3 protein, Hepatitis E virus; Two-Hybrid System Techniques