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Cell Rep. 2014 May 22;7(4):1211-26. doi: 10.1016/j.celrep.2014.03.077. Epub 2014 May 10.

Regulation of FAS exon definition and apoptosis by the Ewing sarcoma protein.

Author information

1
Centre for Genomic Regulation, carrer Doctor Aiguader 88, 08003 Barcelona, Spain; University of Rome "Foro Italico," Piazza Lauro de Bosis 6, 00135 Rome, Italy; Laboratories of Cellular and Molecular Neurobiology and of Neuroimmunology, Fondazione Santa Lucia, 00143 Rome, Italy. Electronic address: mariapaola.paronetto@uniroma4.it.
2
Universitat Pompeu Fabra, carrer Doctor Aiguader 88, 08003 Barcelona, Spain.
3
Laboratories of Cellular and Molecular Neurobiology and of Neuroimmunology, Fondazione Santa Lucia, 00143 Rome, Italy.
4
Centre for Genomic Regulation, carrer Doctor Aiguader 88, 08003 Barcelona, Spain; Universitat Pompeu Fabra, carrer Doctor Aiguader 88, 08003 Barcelona, Spain.
5
Universitat Pompeu Fabra, carrer Doctor Aiguader 88, 08003 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats, Pg Lluis Companys 23, 08010 Barcelona, Spain.
6
Centre for Genomic Regulation, carrer Doctor Aiguader 88, 08003 Barcelona, Spain; Universitat Pompeu Fabra, carrer Doctor Aiguader 88, 08003 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats, Pg Lluis Companys 23, 08010 Barcelona, Spain. Electronic address: juan.valcarcel@crg.eu.

Abstract

The Ewing sarcoma protein EWS is an RNA and DNA binding protein implicated in transcription, pre-mRNA splicing, and DNA damage response. Using CLIP-seq, we identified EWS RNA binding sites in exonic regions near 5' splice sites. A prominent target was exon 6 of the FAS/CD95 receptor, which is alternatively spliced to generate isoforms with opposing activities in programmed cell death. Depletion and overexpression experiments showed that EWS promotes exon 6 inclusion and consequently the synthesis of the proapoptotic FAS/CD95 isoform, whereas an EWS-FLI1 fusion protein characteristic of Ewing sarcomas shows decreased activity. Biochemical analyses revealed that EWS binding promotes the recruitment of U1snRNP and U2AF65 to the splice sites flanking exon 6 and therefore exon definition. Consistent with a role for EWS in the regulation of programmed cell death, cells depleted of EWS show decreased sensitivity to FAS-induced apoptosis, and elevated EWS expression enhances apoptosis in EWS-haploinsufficient Ewing sarcoma cells.

PMID:
24813895
DOI:
10.1016/j.celrep.2014.03.077
[Indexed for MEDLINE]
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