University of Massachusetts Medical School Faculty Publications

UMMS Affiliation

Program in Systems Biology; Program in Molecular Medicine; Department of Biochemistry and Molecular Pharmacology

Publication Date

2016-09-12

Document Type

Article Preprint

Disciplines

Amino Acids, Peptides, and Proteins | Fungi | Genetic Phenomena | Molecular Biology | Nucleic Acids, Nucleotides, and Nucleosides

Abstract

Interactions between RNA binding protein (RBP) and mRNAs are critical to post-transcriptional gene regulation. Eukaryotic genomes encode thousands of mRNAs and hundreds of RBPs. However, in contrast to interactions between transcription factors (TFs) and DNA, the interactome between RBPs and RNA has been explored for only a small number of proteins and RNAs. This is largely because the focus has been on using 'protein-centered' (RBP-to-RNA) interaction mapping methods that identify the RNAs with which an individual RBP interacts. While powerful, these methods cannot as of yet be applied to the entire RBPome. Moreover, it may be desirable for a researcher to identify the repertoire of RBPs that can interact with an mRNA of interest - in a 'gene-centered' manner, yet few such techniques are available. Here, we present Protein-RNA Interaction Mapping Assay (PRIMA) with which an RNA 'bait' can be tested versus multiple RBP 'preys' in a single experiment. PRIMA is a translation-based assay that examines interactions in the yeast cytoplasm, the cellular location of mRNA translation. We show that PRIMA can be used with small RNA elements, as well as with full-length Caenorhabditis elegans 3'UTRs. PRIMA faithfully recapitulates numerous well-characterized RNA-RBP interactions and also identified novel interactions, some of which were confirmed in vivo. We envision that PRIMA will provide a complementary tool to expand the depth and scale with which the RNA-RBP interactome can be explored.

Keywords

molecular biology, Protein-RNA Interaction Mapping Assay, PRIMA, RNA binding protein, mRNA, translation-based assay, yeast cytoplasm

Rights and Permissions

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-ND 4.0 International license.

DOI of Published Version

10.1101/074823

Source

bioRxiv 074823; doi: https://doi.org/10.1101/074823. Link to preprint on bioRxiv service.

Related Resources

Now published in Translation doi: 10.1080/21690731.2017.1295130.

Journal/Book/Conference Title

bioRxiv

Creative Commons License

Creative Commons Attribution-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-No Derivative Works 4.0 License.

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