Multiple CArG boxes in the human cardiac actin gene promoter required for expression in embryonic cardiac muscle cells developing in vitro from embryonal carcinoma cells
Multiple CArG boxes in the human cardiac actin gene promoter required for expression in embryonic cardiac muscle cells developing in vitro from embryonal carcinoma cells
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人心脏肌动蛋白基因启动子中的多个 CArG 盒是在胚胎癌细胞体外发育的胚胎心肌细胞中表达所需的
DOI：
10.1128/mcb.11.9.4796-4803.1991
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发表时间：
1991
影响因子：
5.3
通讯作者：
M. McBurney
中科院分区：
文献类型：
--
作者：
G. Pari;K. Jardine;M. McBurney
构建了人心脏肌动蛋白启动子驱动大肠杆菌lacZ报告基因的嵌合基因，转染并稳定整合到P19胚胎癌细胞基因组中。转染的构建体在二甲亚砜（DMSO）诱导的细胞分化后形成的心肌细胞中表达活跃，但在未分化的培养物和维甲酸处理的培养物中表达不佳，以发育为神经外胚层的衍生物。构建并测试了启动子的一些缺失。在p19来源的心肌中鉴定了有效表达所需的三个区域，每个区域包含被称为CArG盒（CC[AT-rich]6GG）的序列。该分析表明，对心肌表达重要的调控序列存在于之前在骨骼肌母细胞中通过瞬时测定确定的核心启动子的上游。心肌肌动蛋白启动子的表达在myoD蛋白存在的未分化P19细胞中增强了10倍。在p19衍生的心肌细胞中，对表达重要的启动子区域与对肌促性增强重要的启动子区域相似，我们认为这一结果与心肌可能含有肌促性活动的观点一致。
Chimeric genes composed of the human cardiac actin promoter driving the Escherichia coli lacZ reporter gene were constructed, transfected, and stably integrated into genomes of P19 embryonal carcinoma cells. The transfected constructs were expressed actively in cardiac myocytes formed following dimethyl sulfoxide (DMSO)-induced cell differentiation but poorly in undifferentiated cultures and in cultures treated with retinoic acid to develop into derivatives of the neuroectoderm. A number of deletions of the promoter were constructed and tested. Three regions required for efficient expression in P19-derived cardiac muscle were identified, each containing sequences referred to as CArG boxes (CC[AT-rich]6GG). This analysis indicated that regulatory sequences important for expression in cardiac muscle were present upstream of the core promoter identified previously by transient assays in skeletal myoblasts. Expression of the cardiac actin promoter was enhanced 10-fold in undifferentiated P19 cells in the presence of the myoD protein. The promoter regions important for expression in P19-derived cardiocytes were similar to those important for myoD-induced enhancement, a result we interpret to be consistent with the idea that cardiac muscle might contain a myoD-like activity.