Leibham D, Wong M W, Cheng T C, Schroeder S, Weil P A, Olson E N, Perry M
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Mol Cell Biol. 1994 Jan;14(1):686-99. doi: 10.1128/mcb.14.1.686-699.1994.
Members of the MyoD family of helix-loop-helix proteins control expression of the muscle phenotype by regulating the activity of subordinate genes. To investigate processes that control the expression of myogenic factors and regulate the establishment and maintenance of the skeletal muscle phenotype, we have analyzed sequences necessary for transcription of the maternally expressed Xenopus MyoD (XMyoD) gene. A 3.5-kb DNA fragment containing the XMyoDa promoter was expressed in a somite-specific manner in injected frog embryos. The XMyoDa promoter was active in oocytes and cultured muscle cells but not in fibroblasts or nonmuscle cell lines. A 58-bp fragment containing the transcription initiation site, a GC-rich region, and overlapping binding sites for the general transcription factor TFIID and the muscle-specific factor MEF2 was sufficient for muscle-specific transcription. Transcription of the minimal XMyoDa promoter in nonmuscle cells was activated by expression of Xenopus MEF2 (XMEF2) and required binding of both MEF2 and TFIID to the TATA motif. These results demonstrate that the XMyoDa TATA motif is a target for a cell-type-specific regulatory factor and suggests that MEF2 stabilizes and amplifies XMyoDa transcription in mesodermal cells committed to the muscle phenotype.
螺旋-环-螺旋蛋白MyoD家族的成员通过调节下游基因的活性来控制肌肉表型的表达。为了研究控制生肌因子表达以及调节骨骼肌表型建立和维持的过程,我们分析了母源表达的非洲爪蟾MyoD(XMyoD)基因转录所需的序列。一个包含XMyoDa启动子的3.5kb DNA片段在注射的蛙胚胎中以体节特异性的方式表达。XMyoDa启动子在卵母细胞和培养的肌肉细胞中具有活性,但在成纤维细胞或非肌肉细胞系中无活性。一个包含转录起始位点、富含GC区域以及通用转录因子TFIID和肌肉特异性因子MEF2重叠结合位点的58bp片段足以实现肌肉特异性转录。非肌肉细胞中最小XMyoDa启动子的转录可通过非洲爪蟾MEF2(XMEF2)的表达而激活,并且需要MEF2和TFIID都与TATA基序结合。这些结果表明,XMyoDa的TATA基序是细胞类型特异性调节因子的作用靶点,并提示MEF2在致力于形成肌肉表型的中胚层细胞中稳定并增强XMyoDa的转录。