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首页> 外文期刊>Molecular and Cellular Biology >Both muscle-specific and ubiquitous nuclear factors are required for muscle-specific expression of the myosin heavy-chain beta gene in cultured cells.
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Both muscle-specific and ubiquitous nuclear factors are required for muscle-specific expression of the myosin heavy-chain beta gene in cultured cells.

机译：肌肉特异性和普遍存在的核因子都是肌球蛋白重链β基因在培养细胞中的肌肉特异性表达所必需的。

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摘要

Expression of the myosin heavy-chain beta gene is controlled by multiple cis-acting regulatory elements in the 5' flanking region; two of these, referred to as A (-276 to -263) and B (-207 to -180), are essential for conferring muscle-specific activation on homologous and heterologous promoters. Here we report on the identification of nuclear protein factors that specifically bind to these two elements. By using the A element as a probe, as well as nuclear extracts from muscle cells, we found two protein-DNA complexes that displayed distinct bands in a gel mobility shift assay but had identical methylation interference patterns. One complex was present mainly in nuclear extracts from undifferentiated muscle and nonmuscle cells, whereas the other was observed mainly in nuclear extracts from differentiated muscle cells. Thus, the muscle-specific complex formation with the A element appears to be involved in determining tissue-specific expression. Furthermore, competition analysis demonstrated that the A-element-binding factors also bind to the muscle-CAT motif in the cardiac troponin T gene. By using the B element as a probe, we saw similar patterns of gel-shifted bands and methylation interference in nonmuscle and muscle nuclear extracts. In addition, both elements A and B were found to be necessary for tissue-specific expression, suggesting that the muscle-specific activation of the myosin heavy-chain beta gene may require interaction between a muscle-specific and a ubiquitous protein-DNA complex.
机译：肌球蛋白重链β基因的表达受5'侧翼区域中多个顺式作用调控元件的控制。其中两个，分别称为A（-276至-263）和B（-207至-180），对于在同源和异源启动子上赋予肌肉特异性激活至关重要。在这里，我们报告鉴定与这两个元素特异性结合的核蛋白因子。通过使用A元素作为探针以及肌肉细胞的核提取物，我们发现了两种蛋白质-DNA复合物，它们在凝胶迁移率变动分析中显示出不同的条带，但具有相同的甲基化干扰模式。一种复合物主要存在于未分化的肌肉和非肌肉细胞的核提取物中，而另一种复合物则主要存在于分化的肌肉细胞的核提取物中。因此，具有A元素的肌肉特异性复合物形成似乎参与确定组织特异性表达。此外，竞争分析表明，A元素结合因子也与心脏肌钙蛋白T基因中的肌肉CAT基序结合。通过使用B元素作为探针，我们在非肌肉和肌肉核提取物中观察到了相似的凝胶移位带和甲基化干扰模式。另外，发现元素A和B对于组织特异性表达都是必需的，这表明肌球蛋白重链β基因的肌肉特异性活化可能需要肌肉特异性和普遍存在的蛋白质-DNA复合物之间的相互作用。

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