Author/Authors :
Shen, Quan School of Agriculture and Biology - Shanghai JiaoTong University - Shanghai, China , Zhang, Wen School of Medical Technology - Jiangsu University - Zhenjiang - Jiangsu, China , Kang, Yanjun Department of Zoonosis - Institute for Communicable Disease Control and Prevention - Chinese Center for Disease Control and Prevention - Beijing, China , Chen, Yan School of Agriculture and Biology - Shanghai JiaoTong University - Shanghai, China , Cui, Li School of Agriculture and Biology - Shanghai JiaoTong University - Shanghai, China , Yang, Zhibiao School of Agriculture and Biology - Shanghai JiaoTong University - Shanghai, China , Hua, Xiuguo School of Agriculture and Biology - Shanghai JiaoTong University - Shanghai, China
Abstract :
Background: Hepatitis E virus (HEV) is a major causative agent of acute clinical hepatitis
in adults throughout much of Asia, the Middle East, and Africa. The lack of an efficient
cell culture system for HEV has greatly limited our understanding of the mechanisms
of infection, replication, and pathogenicity of this virus. The yeast two-hybridization
system is considered to be an efficient method for determining protein-protein interactions
and screening interactive proteins associated with host cells.
Objectives: In order to identify the host-cell proteins interacting with the HEV-capsid proteins,
a fragment of the HEV-capsid protein p239 (amino acids 368–606) was used as bait;
human liver cDNA library was used as a source of host-cell proteins, and the screening
was performed using the CytoTrap yeast two-hybrid system.
Materials and Methods: The CytoTrap yeast two-hybrid system, which is also called Sos Recruitment
System (SRS), was used to analyze the interaction of the p239 fragment with
host-cell proteins.
Results: We isolated 2 proteins, cytochrome P4502C8 (CYP4502C8) and retinol-binding
protein 4 (RBP4) after 2 rounds of screening. Co-immunoprecipitation assays showed
that both the proteins could bind in vitro to the HEV virion in HepG2 cells.
Conclusions: CYP4502C8 and RBP4 screened from liver cDNA library using the CytoTrap
yeast two-hybrid system interact with HEV capsid in vitro.
