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Originally published In Press as doi:10.1074/jbc.M105642200 on August 20, 2001

J. Biol. Chem., Vol. 276, Issue 44, 40638-40646, November 2, 2001
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TIA-1 and TIAR Activate Splicing of Alternative Exons with Weak 5' Splice Sites followed by a U-rich Stretch on Their Own Pre-mRNAs*

Caroline Le GuinerDagger §, Fabrice Lejeune§||, Delphine GalianaDagger , Liliane Kister, Richard BreathnachDagger , James Stévenin, and Fabienne Del Gatto-KonczakDagger **

From the Dagger  INSERM U463, Institut de Biologie-CHR, 9 Quai Moncousu, 44093 Nantes Cedex 1, France and the  Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, 67404 Illkirch Cedex, France

TIA-1 has recently been shown to activate splicing of specific pre-mRNAs transcribed from transiently transfected minigenes, and of some 5' splice sites in vitro, but has not been shown to activate splicing of any endogenous pre-mRNA. We show here that overexpression of TIA-1 or the related protein TIAR has little effect on splicing of several endogenous pre-mRNAs containing alternative exons, but markedly activates splicing of some normally rarely used alternative exons on the TIA-1 and TIAR pre-mRNAs. These exons have weak 5' splice sites followed by U-rich stretches. When the U-rich stretch following the 5' splice site of a TIA-1 alternative exon was deleted, TIAR overexpression induced use of a cryptic 5' splice site also followed by a U-rich stretch in place of the original splice site. Using in vitro splicing assays, we have shown that TIA-1 is directly involved in activating the 5' splice sites of the TIAR alternative exons. Activation requires a downstream U-rich stretch of at least 10 residues. Our results confirm that TIA-1 activates 5' splice sites followed by U-rich sequences and show that TIAR exerts a similar activity. They suggest that both proteins may autoregulate their expression at the level of splicing.

* This work was supported by funds from the INSERM, the CNRS, the Hôpitaux Universitaires de Nantes et de Strasbourg, the Association pour la Recherche sur le Cancer and the Ligue Nationale contre le Cancer, Comité Departemental de Loire-Atlantique.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.

§ These authors contributed equally.

|| Supported by a fellowship from the Ligue Nationale contre le Cancer.

** To whom correspondence should be addressed: INSERM U463, Institut de Biologie-CHR, 9 Quai Moncousu, 44093 Nantes Cedex 1, France. Tel.: 33-2-40-08-47-50; Fax: 33-2-40-35-66-97; E-mail: fdelgatto@nantes.inserm.fr.

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