α-tropomyosin gene expression in Xenopus laevis: Differential promoter usage during development and controlled expression by myogenic factors

Abstract

 Tropomyosins (TMs) constitute a group of contractile proteins encoded by a multigene family showing distinct cell-type-specific and developmental expression patterns. In mammals and birds, the α-TM gene is the most complex and can produce several muscle and non-muscle isoforms. We report here the characterization of the 5′ region of the Xenopus laevisα-TM gene and its developmental expression. The 5′ region of the gene is structurally related to the avian and mammalian cognates and presents two promoters flanking a pair of alternatively spliced exons, 2a/2b, where exon 2a is a smooth-muscle-specific exon. The internal promoter is used to generate a non-muscle low molecular weight TM whilst muscle TM isoforms originate from the distal promoter. RNase protection analysis shows that the two promoters have distinct temporal programs of activation. The internal promoter is activated early in oogenesis and non-muscle transcripts are found throughout oogenesis, embryogenesis and in adult tissues. Only low molecular weight non-muscle TM-encoding mRNAs are expressed in oogenesis. The distal promoter is silent during oogenesis, and the skeletal muscle α-TM transcripts accumulate from stage 15 in the embryo and are expressed in adult striated muscle tissues. In situ hybridization indicates that these transcripts are expressed in both the somites and heart of the embryo. Ectopic expression of myogenic factors, but not the MEF2 myocyte-specific enhancer factor 2 factors SL1 and SL2, can induce the expression of the α-TM gene suggesting that the gene is a direct target for myogenic but not for MEF2 factors. The amphibian α-TM gene constitutes a gene marker for studying the developmental control expression of muscle genes in the different myogenic lineages.