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Identification of microRNA Precursor-Associated Proteins
Thomas Treiber1, Nora Treiber1, Gunter Meister2
1Laboratory for RNA Biology, Biochemistry Center Regensburg (BZR), University of Regensburg, Regensburg, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|July 1, 2018
Summary
Researchers identified RNA binding proteins (RBPs) that control microRNA (miRNA) production. These proteins bind to specific miRNA structures, influencing the levels of functional miRNAs in cells.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- MicroRNA (miRNA) biogenesis is a complex process regulated at multiple stages.
- Both transcriptional control and post-transcriptional processing influence mature miRNA levels.
- The structure of precursor miRNAs (pri-miRNAs) and their interaction with proteins are critical for efficient processing by Drosha and Dicer.
Purpose of the Study:
- To investigate the role of RNA binding proteins (RBPs) in regulating miRNA biogenesis.
- To identify specific sequence motifs on pri-miRNAs that are recognized by regulatory proteins.
- To develop methods for discovering and analyzing protein regulators of miRNA processing.
Main Methods:
- Utilized pulldown assays with immobilized pri-miRNA hairpins as bait.
- Employed immunoblotting and mass spectrometry for the identification and analysis of bound RBPs.
- Investigated sequence and structural features of pri-miRNA stem loops influencing processing efficiency.
Main Results:
- Demonstrated that specific sequence motifs on pri-miRNAs are recognized by cognate RBPs.
- Showed that these RBPs can modulate the processing efficiency of pri-miRNAs by Drosha and Dicer.
- Successfully isolated and identified protein regulators of miRNA biogenesis using the pulldown approach.
Conclusions:
- RNA binding proteins play a significant role in fine-tuning miRNA biogenesis.
- The interaction between RBPs and pri-miRNA structures provides a mechanism for regulating functional miRNA levels.
- The described pulldown method is effective for discovering novel regulators of miRNA processing.
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