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• W2133148978 endingPage "e0116749" @default.
• W2133148978 startingPage "e0116749" @default.
• W2133148978 abstract "MicroRNAs (miRNAs) are critical post-transcriptional regulators of gene expression. Their precursors have a globally A-form helical geometry, which prevents most proteins from identifying their nucleotide sequence. This suggests the hypothesis that local structural features (e.g., bulges, internal loops) play a central role in specific double-stranded RNA (dsRNA) selection from cellular RNA pools by dsRNA binding domain (dsRBD) containing proteins. Furthermore, the processing enzymes in the miRNA maturation pathway require tandem-dsRBD cofactor proteins for optimal function, suggesting that dsRBDs play a key role in the molecular mechanism for precise positioning of the RNA within these multi-protein complexes. Here, we focus on the tandem-dsRBDs of TRBP, which have been shown to bind dsRNA tightly.We present a combination of dsRNA binding assays demonstrating that TRBP binds dsRNA in an RNA-length dependent manner. Moreover, circular dichroism data shows that the number of dsRBD moieties bound to RNA at saturation is different for a tandem-dsRBD construct than for constructs with only one dsRBD per polypeptide, revealing another reason for the selective pressure to maintain multiple domains within a polypeptide chain. Finally, we show that helical defects in precursor miRNA alter the apparent dsRNA size, demonstrating that imperfections in RNA structure influence the strength of TRBP binding.We conclude that TRBP is responsible for recognizing structural imperfections in miRNA precursors, in the sense that TRBP is unable to bind imperfections efficiently and thus is positioned around them. We propose that once positioned around structural defects, TRBP assists Dicer and the rest of the RNA-induced silencing complex (RISC) in providing efficient and homogenous conversion of substrate precursor miRNA into mature miRNA downstream." @default.
• W2133148978 created "2016-06-24" @default.
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• W2133148978 date "2015-01-21" @default.
• W2133148978 modified "2023-09-23" @default.
• W2133148978 title "Helical Defects in MicroRNA Influence Protein Binding by TAR RNA Binding Protein" @default.
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• W2133148978 doi "https://doi.org/10.1371/journal.pone.0116749" @default.
• W2133148978 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/4301919" @default.
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• W2133148978 hasPublicationYear "2015" @default.
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