Abstract | BACKGROUND: Liver-specific microRNA (miR)-122 has been shown to be involved in regulating translation of hepatitis C viral (HCV) RNA. This study aimed to explore the molecular mechanism of miR-122 in regulating HCV RNA translation initiation. MATERIAL/METHODS: In human liver hepatocellular carcinoma cell line HepG2, UV cross-link assay was performed on a large scale to identify RNA-binding proteins with gradient concentrations of miR-122. Analytical ultracentrifugation was then used to separate the translation initiation complexes. All RNA-binding proteins were then identified by Western blotting. RESULTS: The binding of 68 kDa protein (p68) to HCV RNA was suppressed by the addition of miR-122 via the competitive binding assay. Such inhibition can be eliminated by the addition of 2'-O-methylated oligonucleotides. This binding suppression was determined to be specific for miR-122, which used the mature single-stranded RNA to suppress the binding of p68 onto HCV RNA. This binding inhibition was further validated by using authentic miR-122 with conserved regions and mutated sequences. CONCLUSIONS: The binding of p68 onto HCV RNA can be specifically inhibited by miR-122 via a competitive binding process.
|
Authors | Fu-Tao Zhao, Yun Zhou, Yong-Xing Zhou, Qun Yang, Le Song, Xiao-Jing Jiang, Zhan-Sheng Jia |
Journal | Medical science monitor : international medical journal of experimental and clinical research
(Med Sci Monit)
Vol. 21
Pg. 980-6
(Apr 03 2015)
ISSN: 1643-3750 [Electronic] United States |
PMID | 25836383
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
|
Chemical References |
- 5' Untranslated Regions
- MIRN122 microRNA, human
- MicroRNAs
- RNA, Viral
- RNA-Binding Proteins
|
Topics |
- 5' Untranslated Regions
- Binding, Competitive
- Hep G2 Cells
- Hepacivirus
(genetics, metabolism)
- Humans
- MicroRNAs
(genetics, metabolism)
- Molecular Weight
- Peptide Chain Initiation, Translational
- Protein Binding
- RNA, Viral
(genetics, metabolism)
- RNA-Binding Proteins
(chemistry, metabolism)
|