The epithelial splicing factors ESRP1 and ESRP2 positively and negatively regulate diverse types of alternative splicing events.
Cell-type and tissue-specific alternative splicing events are regulated by combinatorial control involving both abundant RNA binding proteins as well as those with more discrete expression and specialized functions. Epithelial Splicing Regulatory Proteins 1 and 2 (ESRP1 and ESRP2) are recently discovered epithelial-specific RNA binding proteins that promote splicing of the epithelial variant of the FGFR2, ENAH, CD44 and CTNND1 transcripts. To catalogue a larger set of splicing events under the regulation of the ESRPs we profiled splicing changes induced by RNA interference-mediated knockdown of ES RP1 and ES RP2 expression in a human epithelial cell line using the splicing sensitive Affymetrix Exon ST1.0 Arrays. Analysis of the microarray data resulted in the identification of over a hundred candidate ESRP regulated splicing events. We were able to independently validate 38 of these targets by RT-PCR. The ESRP regulated events encompass all known types of alternative splicing events, most prominent being alternative cassette exons and splicing events leading to alternative 3' terminal exons. Importantly, a number of these regulated splicing events occur in gene transcripts that encode proteins with well-described roles in the regulation of actin cytoskeleton organization, cell-cell adhesion, cell polarity and cell migration. In sum, this work reveals a novel list of transcripts differentially spliced in epithelial and mesenchymal cells, implying that coordinated alternative splicing plays a critical role in determination of cell type identity. These results further establish ESRP1 and ESRP2 as global regulators of an epithelial splicing regulatory network.
Warzecha
Claude C
CC
Renal Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA, USA.
Shen
Shihao
S
Xing
Yi
Y
Carstens
Russ P
RP
D017398:Alternative Splicing; D002478:Cells, Cultured; D004847:Epithelial Cells; D006801:Humans; D059630:Mesenchymal Stem Cells; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger; D016601:RNA-Binding Proteins / Q000235:genetics*
C539364:ESRP1 protein, human; C539365:ESRP2 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins