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A Compendium of RNA-Binding Proteins that Regulate MicroRNA Biogenesis.

Molecular cell | 2017

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 ∼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.

Pubmed ID: 28431233 RIS Download

Associated grants

None

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.

This is a list of tools and resources that we have found mentioned in this publication.


Gene Ontology (tool)

RRID:SCR_002811

Computable knowledge regarding functions of genes and gene products. GO resources include biomedical ontologies that cover molecular domains of all life forms as well as extensive compilations of gene product annotations to these ontologies that provide largely species-neutral, comprehensive statements about what gene products do. Used to standardize representation of gene and gene product attributes across species and databases.

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Microsoft Excel (tool)

RRID:SCR_016137

Software application with data analysis tools and spreadsheet templates to track and visualize data. It is used to manage and process data.

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biomaRt (tool)

RRID:SCR_019214

Software package that integrates BioMart data resources with data analysis software in Bioconductor. Can annotate range of gene or gene product identifiers including Entrez Gene and Affymetrix probe identifiers with information such as gene symbol, chromosomal coordinates, Gene Ontology and OMIM annotation. Enables retrieval of genomic sequences and single nucleotide polymorphism information, which can be used in data analysis.

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IRDye 800CW Goat anti-Rabbit IgG (antibody)

RRID:AB_621843

This polyclonal secondary targets IgG

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DDX21 Antibody (antibody)

RRID:AB_513603

This unknown targets DDX21

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ZNF346 antibody (antibody)

RRID:AB_10678096

This polyclonal targets ZNF346 antibody

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HNRNPA1-human (antibody)

RRID:AB_627729

This monoclonal targets HNRNPA1

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Lin28 antibody (antibody)

RRID:AB_776033

This polyclonal targets Human Lin28

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RBM10 antibody (antibody)

RRID:AB_1270200

This polyclonal targets Human RBM10

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NT2-D1 (cell line)

RRID:CVCL_3407

Cell line NT2-D1 is a Cancer cell line with a species of origin Homo sapiens

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NCBI (nonprofit organization)

RRID:SCR_006472

A portal to biomedical and genomic information. NCBI creates public databases, conducts research in computational biology, develops software tools for analyzing genome data, and disseminates biomedical information for the better understanding of molecular processes affecting human health and disease.

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FlyBase (data repository)

RRID:SCR_006549

Database of Drosophila genetic and genomic information with information about stock collections and fly genetic tools. Gene Ontology (GO) terms are used to describe three attributes of wild-type gene products: their molecular function, the biological processes in which they play a role, and their subcellular location. Additionally, FlyBase accepts data submissions. FlyBase can be searched for genes, alleles, aberrations and other genetic objects, phenotypes, sequences, stocks, images and movies, controlled terms, and Drosophila researchers using the tools available from the "Tools" drop-down menu in the Navigation bar.

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miRBase (data repository)

RRID:SCR_003152

Central online repository for microRNA nomenclature, sequence data, annotation and target prediction.Collection of published miRNA sequences and annotation.

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BioMart Project (software resource)

RRID:SCR_002987

THIS RESOURCE IS NO LONGER IN SERVICE. Documented on January 4,2023.Platform provides free software and data services to international scientific community in order to foster scientific collaboration and facilitate scientific discovery process. Project adheres to open source philosophy that promotes collaboration and code reuse.

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UniProt (data or information resource)

RRID:SCR_002380

Collection of data of protein sequence and functional information. Resource for protein sequence and annotation data. Consortium for preservation of the UniProt databases: UniProt Knowledgebase (UniProtKB), UniProt Reference Clusters (UniRef), and UniProt Archive (UniParc), UniProt Proteomes. Collaboration between European Bioinformatics Institute (EMBL-EBI), SIB Swiss Institute of Bioinformatics and Protein Information Resource. Swiss-Prot is a curated subset of UniProtKB.

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Mascot (software resource)

RRID:SCR_014322

A software package and server used to identify and characterize proteins from primary sequence databases using mass spectrometry data. Mascot integrates peptide mass fingerprinting, sequence querying, and MS/MS ion searching in order to search for proteins in databases like SwissProt, NCBInr, EMBL EST divisions, contaminants, and cRAP. If a license is purchased, users may: search data sets that exceed the 1200 spectrum limit of the free version; set up automated, high throughput work; add and edit proteins and quantification methods; and search a preferred collection of sequence databases. The software package works with instruments from AB Sciex, Agilent, Bruker, Jeol, Shimadzu, Thermo Scientific, and Waters.

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MaxQuant (software resource)

RRID:SCR_014485

A quantitative proteomics software package for analyzing large-scale mass-spectrometric data sets. It is a set of algorithms that include peak detection and scoring of peptides, mass calibration, database searches for protein identification, protein quantification, and provides summary statistics.

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Scaffold Proteome Software (software resource)

RRID:SCR_014345

Software for MS/MS proteomic experiments to compare samples, identify biological relevance, and identify isoforms and protein PTMs. These proteins can be classified based on molecular function or organelle. Users can investigate spectrum details and counts, as well as use high through-put batch processing. Tutorials and a free trial are available through the main site.

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Raji (cell line)

RRID:CVCL_0511

Cell line Raji is a Cancer cell line with a species of origin Homo sapiens (Human)

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DU145 (cell line)

RRID:CVCL_0105

Cell line DU145 is a Cancer cell line with a species of origin Homo sapiens (Human)

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SK-N-MC (cell line)

RRID:CVCL_0530

Cell line SK-N-MC is a Cancer cell line with a species of origin Homo sapiens (Human)

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SK-MEL-28 (cell line)

RRID:CVCL_0526

Cell line SK-MEL-28 is a Cancer cell line with a species of origin Homo sapiens (Human)

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A-549 (cell line)

RRID:CVCL_0023

Cell line A-549 is a Cancer cell line with a species of origin Homo sapiens (Human)

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HeLa (cell line)

RRID:CVCL_0030

Cell line HeLa is a Cancer cell line with a species of origin Homo sapiens

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HEK293T (cell line)

RRID:CVCL_0063

Cell line HEK293T is a Transformed cell line with a species of origin Homo sapiens (Human)

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Jurkat E6.1 (cell line)

RRID:CVCL_0367

Cell line Jurkat E6.1 is a Cancer cell line with a species of origin Homo sapiens (Human)

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Hep-G2 (cell line)

RRID:CVCL_0027

Cell line Hep-G2 is a Cancer cell line with a species of origin Homo sapiens (Human)

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LN-229 (cell line)

RRID:CVCL_0393

Cell line LN-229 is a Cancer cell line with a species of origin Homo sapiens (Human)

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DLD-1 (cell line)

RRID:CVCL_0248

Cell line DLD-1 is a Cancer cell line with a species of origin Homo sapiens (Human)

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MCF-7 (cell line)

RRID:CVCL_0031

Cell line MCF-7 is a Cancer cell line with a species of origin Homo sapiens (Human)

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