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

  1. Get@NRC: Competing roles of microRNA-122 recognition elements in hepatitis C virus RNA (Opens in a new window)
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Journal titleVirology
Pages336344; # of pages: 9
SubjectmicroRNA; microRNA 122; unclassified drug; virus RNA; 5' untranslated region; article; binding affinity; controlled study; gene identification; gene interaction; genetic conservation; Hepatitis C virus; human; human cell; in vitro study; interactions with RNA; molecular recognition; nonhuman; nucleotide sequence; open reading frame; priority journal; 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; Humans; MicroRNAs; Mutagenesis, Site-Directed; RNA, Viral; Hepatitis C virus
AbstractMicroRNA-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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AffiliationNational Research Council Canada (NRC-CNRC); NRC Steacie Institute for Molecular Sciences (SIMS-ISSM)
Peer reviewedYes
NPARC number21271598
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Record identifier1fcd2b80-fe5d-441c-a2c7-876e58673b3a
Record created2014-03-24
Record modified2016-05-09
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