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Originally published In Press as doi:10.1074/jbc.M008950200 on April 11, 2001

J. Biol. Chem., Vol. 276, Issue 25, 22317-22322, June 22, 2001
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EWS/FLI Alters 5'-Splice Site Selection*

Lori L. KnoopDagger and Suzanne J. BakerDagger §

From the § Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105 and the Dagger  Department of Pathology, University of Tennessee, Memphis, Tennessee 38163

The chimeric gene EWS/FLI is present in at least 85% of Ewing's sarcomas as a result of chromosomal translocations. The resulting fusion protein contains the N terminus of the RNA-binding protein EWS and the ETS DNA-binding domain of the transcription factor FLI-1. Although EWS/FLI binds DNA and activates transcription, both EWS and EWS/FLI also interact with SF1 and U1C, essential components of the splicing machinery. Therefore, we tested the ability of EWS and EWS/FLI to alter 5'-splice site selection using an E1A gene in vivo splicing assay. We found that EWS/FLI, but not EWS, interfered with heterogeneous nuclear ribonucleoprotein A1-dependent splice site selection of E1A. Mutational analysis of EWS/FLI revealed that the ability to affect pre-mRNA splicing coincided with transforming activity. Therefore, EWS/FLI has the ability to influence splicing as well as transcription.

* This work was supported in part by National Institutes of Health Grant PO1-CA-71907 and Cancer Center Support CORE Grant P30 CA21765 and by the American Lebanese Syrian Associated Charities.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Developmental Neurobiology, St. Jude Children's Research Hospital, 332 North Lauderdale, Memphis, TN 38105. Tel.: 901-495-2254; Fax: 901-495-2270; E-mail: Suzanne.baker@stjude.org.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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