Competing roles of microRNA-122 recognition elements in hepatitis C virus RNA

被引：37

作者：

Nasheri, Neda ^{[1
,2
,3
]}

Singaravelu, Ragunath ^{[1
,2
,3
]}

Goodmurphy, Matthew ^{[1
,2
,3
]}

Lyn, Rodney K. ^{[1
,4
]}

Pezacki, John Paul ^{[1
,2
,3
,4
]}

机构：

[1] Natl Res Council Canada, Steacie Inst Mol Sci, Ottawa, ON K1A 0R6, Canada

[2] Univ Ottawa, Dept Biochem Microbiol & Immunol, Ottawa, ON K1N 6N5, Canada

[3] Univ Ottawa, Ottawa Inst Syst Biol, Ottawa, ON K1N 6N5, Canada

[4] Univ Ottawa, Dept Chem, Ottawa, ON K1N 6N5, Canada

来源：

VIROLOGY | 2011年 / 410卷 / 02期

基金：

加拿大健康研究院; 加拿大自然科学与工程研究理事会;

关键词：

MicroRNA; Hepatitis C; Cooperativity; Translation; RNA replication; Subgenomic replicon; Antiviral immunity; Hybridization kinetics; CIS-ACTING REPLICATION; GENE-EXPRESSION; GENOME; TRANSLATION; SEQUENCES; TARGETS; PROTEIN; REGION; INTERFERENCE; SPECIFICITY;

D O I：

10.1016/j.virol.2010.11.015

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

MicroRNA-122 positively modulates hepatitis C virus (HCV) through direct interactions with viral RNA. Three microRNA-122 recognition elements (MREs) have been previously identified: two in the 5'UTR and one in the 3'UTR. Herein, we report the relative affinity of microRNA-122 to these sites using viral RNA-coated magnetic beads, with mutagenesis and probes to disrupt interactions of microRNA-122 at specific sites. We demonstrate cooperativity in binding between the closely spaced MREs within the 5'UTR in vitro. We also identified a well conserved fourth site in the coding region and showed that it is the highest affinity MRE site. Site-directed mutagenesis of the MREs in HCV subgenomic replicons expressed in Huh-7.5 cells demonstrated competing roles of the stimulatory MREs in the 5'UTR with the inhibitory role of an MRE in the open reading frame (ORF). These data have important implications in elucidating the mechanism of interaction between microRNA-122 and HCV RNA. Crown Copyright (C) 2010 Published by Elsevier Inc. All rights reserved.

引用

页码：336 / 344

页数：9

共 54 条

[21] Position-dependent function for a tandem microRNA miR-122-binding site located in the hepatitis C virus RNA genome [J].