| Objective: To study the interaction between hepatitis C virus nonstructural protein 4A (HCV NS4A) and calcium signal modulating cyclophilin ligand (CAML), identify the site of their interaction and the gene expression change after their interaction.Methods: The proteins interacted with HCV NS4A was screened and identified with yeast two-hybrid in human leukocytes, and the CAML gene was cloned. After the yeast expression vector of CAML was constructed and back-cross was performed, the eukaryotic expression vector of HCV NS4A (pCMV/Myc-NS4A) and CAML (pcDNA3.1/His-A-CAML) was constructed and co-immunoprecipitation was performed in 293 cells. After the yeast expression vectors of different spliceosomes and deletion mutants of CAML gene was constructed and transfected into yeast Y187, matching experiment with AH109 transfected pGBKT7-NS4A was performed. The pCMV/Myc-NS4A was transfected into 293 cells which stable transfected CAML gene, and the gene expression change was detected by microarray test.Results: Fourty-five colonies were sequenced, among which, 29 colonies were CAML. The gene encoding CAML was cloned successfully, and the interaction between NS4A and CAML was verified by back-cross in yeast cells and co-immunoprecipitation in 293 cells. The yeast Y187 transfected the yeast expression vectors of different spliceosomes and deletion mutants of CAML gene could match successfully with AH109 transfected pGBKT7-NS4A. Fourty-eight up-regulated genes and 36 down-regulated genes were detected by microarray test, including 6 genes involved in Rho signal transduction.Conclusions: HCV NS4A could interact with CAML in yeast cells and mammalian cells. The interaction site of HCV NS4A and CAML located at the hydrophilic N-terminal 1-57 amino acid of CAML protein. Maybe, the affection on the Rho signal transduction way, related with apoptosis, could endue cells with resistant to apoptosis via the interaction between HCV NS4A and CAML.
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