A Compendium of RNA-Binding Proteins that Regulate MicroRNA Biogenesis
| Autor: | Simone Harlander, Julia-Lisa Daiß, Kevin Schall, Henning Urlaub, Gerhard Lehmann, Norbert Eichner, Gunter Meister, Thomas Treiber, Uwe Plessmann, Nora Treiber |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
Proteomics
Ribonuclease III 0301 basic medicine Transcription Genetic Ubiquitin-Protein Ligases Regulator RNA-binding protein Computational biology Biology Transfection Interactome DEAD-box RNA Helicases Tripartite Motif Proteins Jurkat Cells Structure-Activity Relationship 03 medical and health sciences RNA interference Databases Genetic microRNA RNA Precursors Humans CRISPR RNA Messenger RNA Processing Post-Transcriptional Molecular Biology Genetics Binding Sites Sequence Analysis RNA High-Throughput Nucleotide Sequencing RNA-Binding Proteins Genomics Hep G2 Cells Cell Biology MicroRNAs HEK293 Cells 030104 developmental biology Gene Expression Regulation A549 Cells RNA splicing MCF-7 Cells Nucleic Acid Conformation RNA Interference CRISPR-Cas Systems Biogenesis HeLa Cells Protein Binding |
| Zdroj: | Molecular Cell |
| ISSN: | 1097-2765 |
| DOI: | 10.1016/j.molcel.2017.03.014 |
| Popis: | During microRNA (miRNA) biogenesis, two endonucleolytic reactions convert stem-loop-structured precursors into mature miRNAs. These processing steps can be posttranscriptionally regulated by RNA-binding proteins (RBPs). Here, we have used a proteomics-based pull-down approach to map and characterize the interactome of a multitude of pre-miRNAs. We identify similar to 180 RBPs that interact specifically with distinct pre-miRNAs. For functional validation, we combined RNAi and CRISPR/Cas-mediated knockout experiments to analyze RBP-dependent changes in miRNA levels. Indeed, a large number of the investigated candidates, including splicing factors and other mRNA processing proteins, have effects on miRNA processing. As an example, we show that TRIM71/LIN41 is a potent regulator of miR-29a processing and its inactivation directly affects miR-29a targets. We provide an extended database of RBPs that interact with pre-miRNAs in extracts of different cell types, highlighting a widespread layer of co- and posttranscriptional regulation of miRNA biogenesis. |
| Databáze: | OpenAIRE |
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