| DOI | Resolve DOI: https://doi.org/10.1016/j.virol.2010.11.015 |
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| Author | Search for: Nasheri, N.1; Search for: Singaravelu, R.1; Search for: Goodmurphy, M.1; Search for: Lyn, R.K.1; Search for: Pezacki, J.P.1 |
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| Affiliation | - National Research Council of Canada. NRC Steacie Institute for Molecular Sciences
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| Format | Text, Article |
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| Subject | microRNA; microRNA 122; unclassified drug; virus RNA; 5' untranslated region; binding affinity; gene identification; gene interaction; genetic conservation; Hepatitis C virus; human cell; in vitro study; interactions with RNA; molecular recognition; nucleotide sequence; open reading frame; replicon; RNA analysis; RNA binding; site directed mutagenesis; translation regulation; virus cell interaction; virus replication; 3' Untranslated Regions; 5' Untranslated Regions; Binding Sites; Cell Line; Hepacivirus; Hepatocytes; MicroRNAs; Mutagenesis, Site-Directed; RNA, Viral; Hepatitis C virus |
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| Abstract | 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. © 2010. |
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| Publication date | 2011 |
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| In | |
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| Language | English |
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| Peer reviewed | Yes |
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| NPARC number | 21271598 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 1fcd2b80-fe5d-441c-a2c7-876e58673b3a |
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| Record created | 2014-03-24 |
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| Record modified | 2020-04-21 |
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