Taylor M V, Gurdon J B, Hopwood N D, Towers N, Mohun T J
Wellcome/Cancer Research Campaign Institute, University of Cambridge.
Genes Dev. 1991 Jul;5(7):1149-60. doi: 10.1101/gad.5.7.1149.
We identify the "M region" of the muscle-specific Xenopus cardiac actin gene promoter from -282 to -348 as necessary for the embryonic expression of a cardiac actin-beta-globin reporter gene injected into fertilized eggs. Four DNA-binding activities in embryo extracts, embryonic M-region factors 1-4 (EMF1-4), are described that interact specifically with this region. One of these, EMF1, is detected in extracts from microdissected somites, which differentiate into muscle, but not in extracts from the adjacent neurectoderm, which differentiates into a variety of other cell types. Moreover, EMF1 is detected in embryo animal caps induced to form mesoderm, which includes muscle, and in which the cardiac actin gene is activated, but not in uninduced animal caps. EMF1 is also first detectable when cardiac actin transcripts begin to accumulate; therefore, both its temporal and spatial distributions during Xenopus development are consistent with a role in activating cardiac actin expression. Two lines of evidence suggest that EMF1 contains the myogenic factor Xenopus MyoD (XMyoD): (1) XMyoD synthesized in vitro can bind specifically to the same site as EMF1; and (2) antibodies raised against XMyoD bind to EMF1. DNA-binding studies indicate that EMF1 may be a complex between XMyoD and proteins found in muscle and other tissues. Our results suggest that the myogenic factor XMyoD, as a component of somite EMF1, regulates the activation of the cardiac actin gene in developing embryonic muscle by binding directly to a necessary region of the promoter.
我们确定了爪蟾肌肉特异性心脏肌动蛋白基因启动子从-282至-348的“M区域”,它对于注入受精卵中的心脏肌动蛋白-β-珠蛋白报告基因的胚胎表达是必需的。本文描述了胚胎提取物中的四种DNA结合活性,即胚胎M区域因子1-4(EMF1-4),它们与该区域特异性相互作用。其中之一,EMF1,在显微解剖的体节提取物中被检测到,体节可分化为肌肉,但在相邻的神经外胚层提取物中未被检测到,神经外胚层可分化为多种其他细胞类型。此外,在诱导形成中胚层(包括肌肉)的胚胎动物帽中检测到EMF1,且心脏肌动蛋白基因在其中被激活,但在未诱导的动物帽中未检测到。当心脏肌动蛋白转录本开始积累时,也首次检测到EMF1;因此,在爪蟾发育过程中其时间和空间分布均与激活心脏肌动蛋白表达的作用一致。有两条证据表明EMF1包含生肌因子爪蟾MyoD(XMyoD):(1)体外合成的XMyoD可与EMF1特异性结合至相同位点;(2)针对XMyoD产生的抗体可与EMF1结合。DNA结合研究表明,EMF1可能是XMyoD与肌肉及其他组织中发现的蛋白质之间形成的复合物。我们的结果表明,生肌因子XMyoD作为体节EMF1的一个组分,通过直接结合启动子的一个必需区域,来调节发育中胚胎肌肉中心脏肌动蛋白基因的激活。