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A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
Published on: June 16, 2022
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Visualizing a protonated RNA state that modulates microRNA-21 maturation
Jared T Baisden1, Joshua A Boyer1, Bo Zhao1,2
1Department of Biochemistry and Biophysics, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Nature Chemical Biology
|October 27, 2020
Summary
MicroRNA maturation is regulated by pH-dependent structural dynamics. The precursor of microRNA-21 uses an alternative conformation to enhance Dicer processing, revealing a new layer of microRNA biogenesis control.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- MicroRNAs (miRNAs) are crucial noncoding RNAs regulating gene expression.
- miRNA biogenesis is a tightly controlled process involving specific protein interactions.
- Understanding miRNA maturation mechanisms is key to deciphering gene regulation.
Purpose of the Study:
- To investigate the structural dynamics of precursor microRNA-21 (pre-miR-21).
- To elucidate the role of these dynamics in Dicer processing and miRNA maturation.
- To explore pH-dependent regulation of miRNA biogenesis.
Main Methods:
- Nuclear Magnetic Resonance (NMR) relaxation dispersion spectroscopy.
- Site-directed mutagenesis.
- Biochemical assays for Dicer processing.
Main Results:
- Pre-miR-21 exists as a pH-dependent dynamic ensemble.
- An alternative conformation involves a protonated adenine-guanine mismatch, sequestering bulged adenine.
- This conformation significantly enhances Dicer enzyme processing efficiency.
Conclusions:
- MicroRNA maturation efficiency can be encoded within the intrinsic dynamics of precursor miRNAs.
- pH-dependent structural ensembles offer a mechanism for regulating miRNA biogenesis.
- These findings provide insights into cellular responses to environmental stimuli.
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