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J. Biol. Chem., Vol. 281, Issue 45, 34406-34420, November 10, 2006

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Transcription Enhancer Factor-1-dependent Expression of the -Tropomyosin Gene in the Three Muscle Cell Types^*

Stéphanie Pasquet¹², François Naye¹², Corinne Faucheux, Odile Bronchain^¶, Albert Chesneau^¶, Pierre Thiébaud³, and Nadine Thézé⁴

From the Unité INSERM 441, Avenue du Haut-Lévêque, 33600 Pessac, France, Unité Mixte de Recherche CNRS 5164, Université Bordeaux 2, 146 rue Léo Saignat, 33076 Bordeaux, France, and ^¶Unité Mixte de Recherche CNRS 8080, Université Paris-Sud, 91405 Orsay, France

In vertebrates, the actin-binding proteins tropomyosins are encoded by four distinct genes that are expressed in a complex pattern during development and muscle differentiation. In this study, we have characterized the transcriptional machinery of the -tropomyosin (-Tm) gene in muscle cells. Promoter analysis revealed that a 284-bp proximal promoter region of the Xenopus laevis -Tm gene is sufficient for maximal activity in the three muscle cell types. The transcriptional activity of this promoter in the three muscle cell types depends on both distinct and common cis-regulatory sequences. We have identified a 30-bp conserved sequence unique to all vertebrate -Tm genes that contains an MCAT site that is critical for expression of the gene in all muscle cell types. This site can bind transcription enhancer factor-1 (TEF-1) present in muscle cells both in vitro and in vivo. In serum-deprived differentiated smooth muscle cells, TEF-1 was redistributed to the nucleus, and this correlated with increased activity of the -Tm promoter. Overexpression of TEF-1 mRNA in Xenopus embryonic cells led to activation of both the endogenous -Tm gene and the exogenous 284-bp promoter. Finally, we show that, in transgenic embryos and juveniles, an intact MCAT sequence is required for correct temporal and spatial expression of the 284-bp gene promoter. This study represents the first analysis of the transcriptional regulation of the -Tm gene in vivo and highlights a common TEF-1-dependent regulatory mechanism necessary for expression of the gene in the three muscle lineages.

Received for publication, March 10, 2006 , and in revised form, September 6, 2006.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) DQ871277 [GenBank] .

^* This work was supported by INSERM, the University of Bordeaux 2, the Association Française contre les Myopathies, the Fondation de France, and the Région Aquitaine. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Both authors contributed equally to this work.

² Supported by a Ph.D. fellowship from the Ministère de l'Éducation Nationale, de la Recherche, et de la Technologie.

³ To whom correspondence may be addressed: Unité INSERM 441, Ave. du Haut Lévêque, 33600 Pessac, France. Tel.: 33-5-5789-0102; Fax: 33-5-5636-8979; E-mail: pierre.thiebaud{at}bordeaux.inserm.fr. ⁴ To whom correspondence may be addressed: Unité INSERM 441, Ave. du Haut Lévêque, 33600 Pessac, France. Tel.: 33-5-5789-1976; Fax: 33-5-5636-8979; E-mail: nadine.theze{at}bordeaux.inserm.fr.

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