Identification of the RNA Recognition Element of the RBPMS Family of RNA-Binding Proteins and Their Transcriptome-Wide Mrna Targets
Downloaded from rnajournal.cshlp.org on October 1, 2021 - Published by Cold Spring Harbor Laboratory Press Identification of the RNA recognition element of the RBPMS family of RNA-binding proteins and their transcriptome-wide mRNA targets THALIA A. FARAZI,1,5 CARL S. LEONHARDT,1,5 NEELANJAN MUKHERJEE,2 ALEKSANDRA MIHAILOVIC,1 SONG LI,3 KLAAS E.A. MAX,1 CINDY MEYER,1 MASASHI YAMAJI,1 PAVOL CEKAN,1 NICHOLAS C. JACOBS,2 STEFANIE GERSTBERGER,1 CLAUDIA BOGNANNI,1 ERIK LARSSON,4 UWE OHLER,2 and THOMAS TUSCHL1,6 1Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, New York 10065, USA 2Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine, 13125 Berlin, Germany 3Biology Department, Duke University, Durham, North Carolina 27708, USA 4Institute of Biomedicine, The Sahlgrenska Academy, University of Gothenburg, Gothenburg, SE-405 30, Sweden ABSTRACT Recent studies implicated the RNA-binding protein with multiple splicing (RBPMS) family of proteins in oocyte, retinal ganglion cell, heart, and gastrointestinal smooth muscle development. These RNA-binding proteins contain a single RNA recognition motif (RRM), and their targets and molecular function have not yet been identified. We defined transcriptome-wide RNA targets using photoactivatable-ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) in HEK293 cells, revealing exonic mature and intronic pre-mRNA binding sites, in agreement with the nuclear and cytoplasmic localization of the proteins. Computational and biochemical approaches defined the RNA recognition element (RRE) as a tandem CAC trinucleotide motif separated by a variable spacer region. Similar to other mRNA-binding proteins, RBPMS family of proteins relocalized to cytoplasmic stress granules under oxidative stress conditions suggestive of a support function for mRNA localization in large and/or multinucleated cells where it is preferentially expressed.
[Show full text]