title: RNA-binding protein RBM20 represses splicing to orchestrate cardiac pre-mRNA processing.
abstract: Mutations in the gene encoding the RNA-binding protein RBM20 have been implicated in dilated cardiomyopathy (DCM), a major cause of chronic heart failure, presumably through altering cardiac RNA splicing. Here, we combined transcriptome-wide crosslinking immunoprecipitation (CLIP-seq), RNA-seq, and quantitative proteomics in cell culture and rat and human hearts to examine how RBM20 regulates alternative splicing in the heart. Our analyses revealed the presence of a distinct RBM20 RNA-recognition element that is predominantly found within intronic binding sites and linked to repression of exon splicing with RBM20 binding near 3' and 5' splice sites. Proteomic analysis determined that RBM20 interacts with both U1 and U2 small nuclear ribonucleic particles (snRNPs) and suggested that RBM20-dependent splicing repression occurs through spliceosome stalling at complex A. Direct RBM20 targets included several genes previously shown to be involved in DCM as well as genes not typically associated with this disease. In failing human hearts, reduced expression of RBM20 affected alternative splicing of several direct targets, indicating that differences in RBM20 expression may affect cardiac function. Together, these findings identify RBM20-regulated targets and provide insight into the pathogenesis of human heart failure.
authors: Henrike
authors: H
authors: Maatz
authors: Marvin
authors: M
authors: Jens
authors: Martin
authors: M
authors: Liss
authors: Sebastian
authors: S
authors: Schafer
authors: Matthias
authors: M
authors: Heinig
authors: Marieluise
authors: M
authors: Kirchner
authors: Eleonora
authors: E
authors: Adami
authors: Carola
authors: C
authors: Rintisch
authors: Vita
authors: V
authors: Dauksaite
authors: Michael H
authors: MH
authors: Radke
authors: Matthias
authors: M
authors: Selbach
authors: Paul J R
authors: PJ
authors: Barton
authors: Stuart A
authors: SA
authors: Cook
authors: Nikolaus
authors: N
authors: Rajewsky
authors: Michael
authors: M
authors: Gotthardt
authors: Markus
authors: M
authors: Landthaler
authors: Norbert
authors: N
authors: Hubner
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002311:Cardiomyopathy, Dilated / Q000150:complications / Q000235:genetics / Q000378:metabolism; D015331:Cohort Studies; D005091:Exons; D006333:Heart Failure / Q000209:etiology / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009154:Mutation; D009206:Myocardium / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D022821:RNA Splice Sites; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000378:metabolism; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000378:metabolism; D012641:Selection, Genetic; D017381:Spliceosomes / Q000378:metabolism
chemical_list: C573837:RBM20 protein, rat; D012322:RNA Precursors; D022821:RNA Splice Sites; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; D017413:Ribonucleoprotein, U2 Small Nuclear; C543725:ribonucleic acid binding motif protein 20, human