Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway

Rongjuan Pei; Xiaoyong Zhang; Song Xu; Zhongji Meng; Michael Roggendorf; Mengji Lu; Xinwen Chen

doi:10.1007/s12250-012-3257-6

October 2012

Rongjuan Pei, Xiaoyong Zhang, Song Xu, Zhongji Meng, Michael Roggendorf, Mengji Lu and Xinwen Chen. Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway[J]. Virologica Sinica, 2012, 27(5): 278-285. doi: 10.1007/s12250-012-3257-6

Citation: Rongjuan Pei, Xiaoyong Zhang, Song Xu, Zhongji Meng, Michael Roggendorf, Mengji Lu, Xinwen Chen. Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway .VIROLOGICA SINICA, 2012, 27(5) : 278-285. http://dx.doi.org/10.1007/s12250-012-3257-6

Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway

cstr: 32224.14.s12250-012-3257-6

1.
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China; 2Institute of Virology, University Hospital of Essen, University of Duisburg-Essen, Essen 45122, Germany; 3Graduate school of the Chinese Academy of Sciences, Beijing 100049, China

收稿日期： 2012-04-26
录用日期： 2012-08-17

摘要

The mitogen activated protein kinases-extracellular signal regulated kinases (MAPK-ERK) pathway is involved in regulation of multiple cellular processes including the cell cycle. In the present study using a Huh7 cell line Con1 with an HCV replicon, we have shown that the MAPK-ERK pathway plays a significant role in the modulation of HCV replication and protein expression and might influence IFN-? signalling. Epithelial growth factor (EGF) was able to stimulate ERK activation and decreased HCV RNA load while a MAPK-ERK pathway inhibitor U0126 led to an elevated HCV RNA load and higher NS5A protein amounts in Con1 cells. It could be demonstrated that the inhibition of the MAPK-ERK pathway facilitated the translation directed by the HCV internal ribosome entry site. Consistently, a U0126 treatment enhanced activity of the HCV reporter replicon in transient transfection assays. Thus, the MAPK-ERK pathway plays an important role in the regulation of HCV gene expression and replication. In addition, cyclin-dependent kinases (CDKs) downstream of ERK may be involved in the modulation of HCV replication since roscovitine, an inhibitor of CDKs had a similar effect to that of U0126. Modulation of the cell cycle progression by cell cycle inhibitor or RNAi resulted consistently in changes of HCV RNA levels. Further, the replication of HCV replicon in Con1 cells was inhibited by IFN-?. The inhibitory effect of IFN-? could be partly reversed by pre-incubation of Con-1 cells with inhibitors of the MAPK-ERK pathway and CDKs. It could be shown that the MAPK-ERK inhibitors are able to partially modulate the expression of interferon-stimulated genes.
- Hepatitis C Virus (HCV)
- , Mitogen activated protein kinases-extracellular signal regulated kinase (MAPK-ERK)
- , Cell cycle progression

Regulation of Hepatitis C Virus Replication and Gene Expression by the MAPK-ERK Pathway

1.
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
2.
Institute of Virology, University Hospital of Essen, University of Duisburg-Essen, Essen 45122, Germany
3.
Graduate School of the Chinese Academy of Sciences, Beijing 100049, China

Corresponding author: Xinwen Chen, chenxw@wh.iov.cn
Received Date: 26 April 2012
Accepted Date: 17 August 2012

Fund Project: Partly supported by a joint grant of Chinese Academy of Science and Deutsche Akademische Austausch Dienst, and the the National Basic Research Priorities Program of China 2005CB522901Partly supported by a joint grant of Chinese Academy of Science and Deutsche Akademische Austausch Dienst, and the the National Basic Research Priorities Program of China 2009CB522501Partly supported by a joint grant of Chinese Academy of Science and Deutsche Akademische Austausch Dienst, and the the National Basic Research Priorities Program of China 2007CB512901

Abstract

The mitogen activated protein kinases-extracellular signal regulated kinases (MAPK-ERK) pathway is involved in regulation of multiple cellular processes including the cell cycle. In the present study using a Huh7 cell line Con1 with an HCV replicon, we have shown that the MAPK-ERK pathway plays a significant role in the modulation of HCV replication and protein expression and might influence IFN-? signalling. Epithelial growth factor (EGF) was able to stimulate ERK activation and decreased HCV RNA load while a MAPK-ERK pathway inhibitor U0126 led to an elevated HCV RNA load and higher NS5A protein amounts in Con1 cells. It could be demonstrated that the inhibition of the MAPK-ERK pathway facilitated the translation directed by the HCV internal ribosome entry site. Consistently, a U0126 treatment enhanced activity of the HCV reporter replicon in transient transfection assays. Thus, the MAPK-ERK pathway plays an important role in the regulation of HCV gene expression and replication. In addition, cyclin-dependent kinases (CDKs) downstream of ERK may be involved in the modulation of HCV replication since roscovitine, an inhibitor of CDKs had a similar effect to that of U0126. Modulation of the cell cycle progression by cell cycle inhibitor or RNAi resulted consistently in changes of HCV RNA levels. Further, the replication of HCV replicon in Con1 cells was inhibited by IFN-?. The inhibitory effect of IFN-? could be partly reversed by pre-incubation of Con-1 cells with inhibitors of the MAPK-ERK pathway and CDKs. It could be shown that the MAPK-ERK inhibitors are able to partially modulate the expression of interferon-stimulated genes.

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