title: TRBP alters human precursor microRNA processing in vitro.
abstract: MicroRNAs play central roles in controlling gene expression in human cells. Sequencing data show that many miRNAs are produced at different levels and as multiple isoforms that can vary in length at their 5' or 3' ends, but the biogenesis and functional significance of these RNAs are largely unknown. We show here that the human trans-activation response (TAR) RNA binding protein (TRBP), a known molecular partner of the miRNA processing enzyme Dicer, changes the rates of pre-miRNA cleavage in an RNA-structure-specific manner. Furthermore, TRBP can trigger the generation of iso-miRNAs (isomiRs) that are longer than the canonical sequence by one nucleotide. We show that this change in miRNA processing site can alter guide strand selection, resulting in preferential silencing of a different mRNA target. These results implicate TRBP as a key regulator of miRNA processing and targeting in humans.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California 94720, USA.
authors.forename: Ho Young
authors.initials: HY
authors.lastname: Lee
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Doudna
mesh_terms: D001483:Base Sequence; D053487:DEAD-box RNA Helicases / Q000737:chemistry / Q000378:metabolism; D017930:Genes, Reporter; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D035683:MicroRNAs / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D059367:RNA Cleavage; D012322:RNA Precursors / Q000737:chemistry* / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D043244:Ribonuclease III / Q000737:chemistry / Q000378:metabolism; D017423:Sequence Analysis, RNA; D013379:Substrate Specificity; D000099253:Trans-Activation Responsive RNA-Binding Protein
chemical_list: D053487:DEAD-box RNA Helicases; D049410:Luciferases, Renilla; D035683:MicroRNAs; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D043244:Ribonuclease III; D000099253:Trans-Activation Responsive RNA-Binding Protein; C408087:DICER1 protein, human