- 作者：Long V. Pham ; Huong T.T. Ngo ; Yun-Sook Lim ; Soon B. Hwang
- 关键词：HCV ; hepatitis C virus ; NS5A ; non-structural 5A ; NS5B ; non-structural 5B ; HCC ; hepatocellular carcinoma ; CycA2 ; cyclin A2 ; CDK1&2 ; cyclin-dependent kinase 1&2 ; GST ; glutathione S-transferase ; PBS ; phosphate-buffered saline ; TRITC ; tetramethylrhodamine iso
- 刊名：Journal of Hepatology
- 出版年：2012
- 出版时间：November, 2012
- 年：2012
- 卷：57
- 期：5
- 页码：960-966
- 全文大小：1006 K
Background & Aims
Hepatitis C virus (HCV) requires host cellular proteins for its own propagation. To identify the cellular factors necessary for HCV propagation, we have recently screened the small interfering RNA (siRNA) library targeting cell cycle genes using cell culture grown HCV (HCVcc)-infected cells. In the current study, we have selected and characterized the gene encoding Cyclin A2 (CycA2). Deregulation of CycA2 has been implicated in many types of cancers, including hepatocellular carcinoma.Methods
The effects of CycA2 on HCV propagation were investigated by siRNA-mediated knockdown assay, in vitro and in vivo protein binding assays, luciferase reporter gene assay, and immunoblot assay.
Results
We showed that siRNA-mediated depletion of CycA2 significantly inhibited HCV replication in both HCV subgenomic replicon cells and HCVcc-infected cells. Furthermore, HCV non-structural 5B (NS5B) specifically interacted with CycA2 in vitro and in vivo. Protein interaction was mediated through the cyclin box of CycA2 and the palm domain of NS5B. We further showed that R/HxL motif in the palm domain of HCV NS5B mediated protein interaction with CycA2 and this interaction was necessary for HCV replication. Moreover, we demonstrated that tylophorine, the natural plant product exerting a CycA2 inhibitory function, abrogated HCV replication.
Conclusions
HCV regulates CycA2 via NS5B protein for its own propagation. In addition, tylophorine may be a potential therapeutic agent for HCV.