Transcription enhancer factor-1-dependent expression of the alpha-tropomyosin gene in the three muscle cell types.
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 alpha-tropomyosin (alpha-Tm) gene in muscle cells. Promoter analysis revealed that a 284-bp proximal promoter region of the Xenopus laevis alpha-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 alpha-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 alpha-Tm promoter. Overexpression of TEF-1 mRNA in Xenopus embryonic cells led to activation of both the endogenous alpha-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 alpha-Tm gene in vivo and highlights a common TEF-1-dependent regulatory mechanism necessary for expression of the gene in the three muscle lineages.
Pasquet
Stéphanie
S
Unité INSERM 441, Avenue du Haut-Lévêque, 33600 Pessac, France.
Naye
François
F
Faucheux
Corinne
C
Bronchain
Odile
O
Chesneau
Albert
A
Thiébaud
Pierre
P
Thézé
Nadine
N
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002467:Cell Nucleus; D002478:Cells, Cultured; D002645:Chickens; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins; D024202:Electrophoretic Mobility Shift Assay; D004742:Enhancer Elements, Genetic / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D008969:Molecular Sequence Data; D032342:Muscle Cells / Q000378:metabolism; D009132:Muscles / Q000196:embryology* / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D032383:Myocytes, Cardiac; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000235:genetics; D014176:Protein Biosynthesis; D011817:Rabbits; D051381:Rats; D000091262:TEA Domain Transcription Factors; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D014335:Tropomyosin / Q000235:genetics* / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology / Q000378:metabolism
D004268:DNA-Binding Proteins; D000091262:TEA Domain Transcription Factors; C083215:Tead1 protein, mouse; D014157:Transcription Factors; D014335:Tropomyosin