JBC Transcription and Nuclear Factor Monoclonals

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Originally published In Press as doi:10.1074/jbc.M008665200 on March 26, 2001

J. Biol. Chem., Vol. 276, Issue 24, 21062-21069, June 15, 2001
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Promoter Choice Influences Alternative Splicing and Determines the Balance of Isoforms Expressed from the Mouse bcl-X Gene*

Adali PecciDagger §, Luciana Rocha Viegas§, José Lino Barañao§, and Miguel BeatoDagger ||

From the Dagger  Institut für Molekularbiologie und Tumorforschung (IMT), Marburg 35033, Germany and § Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, and Instituto de Biología y Medicina Experimental (IByME), Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires 1428, Argentina

Differential splicing from the bcl-X gene generates several isoforms with opposite effects on the apoptotic response. To explore the mechanism controlling the balance between the various isoforms, we have characterized the 5' region of the mouse bcl-X gene. We identified three new promoters in addition to the two previously described (Grillot, D. A., M., G.-G., Ekhterae, D., Duan, L., Inohara, N., Ohta, S., Seldin, M. F., and Núñez, G. (1997) J. Immunol. 158, 4750-4757). These five promoters (P1-P5) would give rise to at least five mRNAs with different 5'-untranslated region, all sharing the same translation initiation site. Except for the product of the most proximal promoter (P1), the other mRNAs are generated by alternative splicing of noncoding exons to a common acceptor site located in the first translated exon. Reverse transcriptase-polymerase chain reaction, primer extension, and RNase protection assays demonstrate a tissue-specific pattern of promoter usage. P1 and P2 are active in all tissues analyzed, whereas the other three promoter show tissue-specific activities. P3 is active in spleen, liver, and kidney, P4 is active in uterus and spleen, and P5 is active in spleen, liver, brain, and thymus. We present evidence suggesting that promoter selection influences the outcome of the splice process. Transcripts from P1 generate mainly the mRNA for the long isoform Bcl-XL, whereas transcripts from P2 generate mRNAs for the isoforms Bcl-XL, Bcl-XS, and Bcl-Xgamma and transcripts from P3 yield mainly mRNAs for the isoform Bcl-Xgamma . Our results suggest a key role of promoter choice in determining alternative splicing and, thus, the balance of Bcl-X isoforms.

* This work was supported by grants from the Deutsche Forschungsgemeinschaft and the Fond der Chemischen Industrie (to M. B.) as well as by Universidad de Buenos Aires para Ciencia y Tecnica (UBACYT) Grant TX 82, Fon dos para la Ciencia y Tecnica Grant PICT 970256, and Proyectos deInvestigacion Plurianuales Grant 4404 (to J. L. B.), UBACYT Grant JX 05 and Programa de Regulacion Hormonal y Metabolica-Consejo Nacional de Investigaciones Científicas y Técnicas (to A. P.), and Fundacion Antorchas-Deutsche Akademische Austauschdienst (to M. B. and J. L. B.).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.

An established researcher from the Consejo Nacional de Investigaciones Científicas y Técnicas.

|| To whom correspondence should be addressed. Tel.: 49-6421-28-6286; Fax: 49-6421-28-5398: E-mail: beato@imt.uni-marburg.de.

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