Cell-specific roles of Domains I and II of HCV 5'untranslated region in the translation initiation activity
10.3969/j.issn.1673-4254.2014.12.25
- VernacularTitle:丙型肝炎病毒5'端非编码区结构域Ⅱ、Ⅱ在其翻译启动活性中的作用具有细胞特异性
- Author:
Xiaoye HUANG
1
;
Lisha LIU
;
Guangjing CUI
;
Xixia LIU
;
Meitong LIU
;
Qiongshan MA
;
Shuiping LIU
Author Information
1. 中南大学湘雅医学院医学微生物学系
- Keywords:
hepatitis C virus;
5' untranslated region;
translation initiation activity;
cell-specific
- From:
Journal of Southern Medical University
2014;(12):1826-1829
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the roles of Domain I and Domain II of hepatitis C virus (HCV) 5' untranslated region (UTR) in the translation initiation activity of HCV 5'UTR in different host cell lines. Methods The eukaryotic expression plasmid pCMVNCRLuc (pCN1), in which full-length HCV 5'UTR regulates firefly luciferase expression, was modified by deleting Domain I and the downstream single-stranded sequence (43 bp in total) from the UTR (pCNl-d2), Domain I with the downstream single-stranded sequence and Domain II (118 bp in total) from the UTR (pCNl-d3), or the total UTR (pCNl-d5). The modified plasmids were transfected via liposome into different cell lines with pRL-TK plasmid co-transfected as the normalization control. At 36 h after the transfection, the total cellular RNA was harvested for semi-quantitative RT-PCR, and the relative expression activities of luciferase were assayed with a dual luciferase reporter gene assay system. The translation initiation activities of the truncated HCV 5'UTRs in different translation systems were analyzed. Results Deletion of Domain I and the downstream single-stranded sequence caused no significant changes of the translational activity of HCV 5'UTR in Hela or C6 cells, but decreased the translational activity by 46%in L-02 cells and increased the translational activity by 46%in 293T cells. Deletion of both Domain I and Domain II resulted in decreased translational activity of HCV 5'UTR by 51%in HeLa cells, but increased the translational activity by 40%in L-02 cells, 60%in C6 cells and 135%in 293T cells. Conclusions Domain I and Domain II of HCV 5'UTR perform cell type-specific roles in HCV IRES-driven translation initiation.
