Competitive binding between miR-122 and p68 onto hepatitis C viral RNA.

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)