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• W2318205280 endingPage "2566" @default.
• W2318205280 startingPage "2553" @default.
• W2318205280 abstract "p54nrb/NonO is a nuclear RNA-binding protein involved in many cellular events such as pre-mRNA processing, transcription, and nuclear retention of hyper-edited RNAs. In particular, it participates in the splicing process by directly binding the 5′ splice site of pre-mRNAs. The protein also concentrates in a nuclear body called paraspeckle by binding a G-rich segment of the ncRNA NEAT1. The N-terminal section of p54nrb/NonO contains tandem RNA recognition motifs (RRMs) preceded by an HQ-rich region including a threonine residue (Thr15) whose phosphorylation inhibits its RNA binding ability, except for G-rich RNAs. In this work, our goal was to understand the rules that characterize the binding of the p54nrb/NonO RRMs to their RNA target. We have done in vitro RNA binding experiments which revealed that only the first RRM of p54nrb/NonO binds to the 5′ splice site RNA. We have then determined the structure of the p54nrb/NonO RRM1 by liquid-state NMR which revealed the presence of a canonical fold (β1α1β2β3α2β4) and the conservation of aromatic amino acids at the protein surface. We also investigated the dynamics of this domain by NMR. The p54nrb/NonO RRM1 displays some motional properties that are typical of a well-folded protein with some regions exhibiting more flexibility (loops and β-strands). Furthermore, we determined the affinity of p54nrb/NonO RRM1 interaction to the 5′ splice site RNA by NMR and fluorescence quenching and mapped its binding interface by NMR, concluding in a classical nucleic acid interaction. This study provides an improved understanding of the molecular basis (structure and dynamics) that governs the binding of the p54nrb/NonO RRM1 to one of its target RNAs." @default.
• W2318205280 created "2016-06-24" @default.
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• W2318205280 date "2016-04-26" @default.
• W2318205280 modified "2023-10-14" @default.
• W2318205280 title "Structure, Dynamics, and Interaction of p54^nrb/NonO RRM1 with 5′ Splice Site RNA Sequence" @default.
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• W2318205280 doi "https://doi.org/10.1021/acs.biochem.5b01240" @default.
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• W2318205280 hasPublicationYear "2016" @default.
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