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Originally published In Press as doi:10.1074/jbc.M210737200 on November 19, 2002

J. Biol. Chem., Vol. 278, Issue 5, 3040-3047, January 31, 2003
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Biochemical Function of Female-Lethal (2)D/Wilms' Tumor Suppressor-1-associated Proteins in Alternative Pre-mRNA Splicing*

Angeles Ortegaab, Martina Niksiccd, Angela Bachief, Matthias Wilme, Lucas Sánchezgh, Nicholas Hastieci, and Juan Valcárcelaj

From the a Gene Expression Programme, European Molecular Biology Laboratory, Heidelberg, Germany, the c Medical Research Council Human Genetics Unit, Edinburgh, United Kingdom, the e Biochemical Instrumentation Programme, EMBL, Heidelberg, Germany, and the g Centro de Investigaciones Biológicas, Madrid, Spain

Genetic and molecular data have implicated the Drosophila gene female-lethal (2)d (fl (2)d) in alternative splicing regulation of genes involved in sexual determination. Sex-specific splicing is under the control of the female-specific regulatory protein sex-lethal (SXL). Co-immunoprecipitation and mass spectrometry results indicate that SXL and FL (2)D form a complex and that the protein VIRILIZER and a Ran-binding protein implicated in protein nuclear import are also present in complexes containing FL (2)D. A human homolog of FL (2)D was identified and cloned. Interestingly, this gene encodes a protein (WTAP) that was previously found to interact with the Wilms' tumor suppressor-1 (WT1), an isoform of which binds to and co-localizes with splicing factors. Alternative splicing of transformer pre-mRNA, a target of SXL regulation, was affected by immunodepletion of hFL (2)D/WTAP from HeLa nuclear extracts, thus arguing for a biochemical function of FL (2)D/WTAP proteins in splicing regulation.

* This work was supported in part by a Human Frontiers Science Program Organization grant (to J. V.).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.

b Supported by fellowships from University of Granada, Ministerio de Educación y Ciencia (Spain) and Marie Curie Research Fellowships Program. Present address: Dept. of Biochemistry and Molecular Biology, Faculty of Pharmacy, University of Granada, Spain.

d Recipient of an EMBO short-term fellowship.

f Present address: Dibit, San Raffaelle Scientific Institute, 20132 Milano, Italy.

h Supported by Dirección General de Investigación Científica y Técnica Grant PB98-0466.

i Supported by the Medical Research Council and the European Union.

j To whom correspondence should be addressed. Present address: Centre de Regulacio Genomica, Passeig Maritim 37-49, 08003 Barcelona, Spain. Tel.: 34-93-2240956; Fax: 34-93-2240899; E-mail: juan.valcarcel@crg.es.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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