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首页> 外文期刊>Molecular and Cellular Biology >Interaction of nuclear proteins with a positive cis-acting element of rat embryonic myosin heavy-chain promoter: identification of a new transcriptional factor.
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Interaction of nuclear proteins with a positive cis-acting element of rat embryonic myosin heavy-chain promoter: identification of a new transcriptional factor.

机译：核蛋白与大鼠胚胎肌球蛋白重链启动子的顺式作用元件的相互作用：鉴定新的转录因子。

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摘要

A DNA fragment of the rat embryonic myosin heavy-chain promoter (MHCemb) has been found to specifically bind a nuclear factor (NFe) present in extracts prepared from mouse C2 myoblasts, myotubes, and HeLa cells. The nucleotide sequence of the binding site (BSe) has been identified as 5'-GTGTCAGTCA-3' and was located between -93 and -84. Transient expression studies on MHCemb promoter deletion constructs in C2 myoblasts and C2 myotubes suggested that NFe is a transcriptional factor. Deletion of the NFe-binding site resulted in four- to sixfold and twofold reduction of promoter activity in C2 myotubes and C2 myoblasts, respectively. Furthermore, point mutations at the BSe not only abolished the NFe-binding activity of the MHCemb promoter but also resulted in reduction of the promoter activity to levels similar to those of the deletion constructs in C2 myotubes, myoblasts, and Hela cells (four- to sixfold). Although BSe and the binding site of the recently identified transcriptional factors AP-1 and ATF share significant homology, the results from competition binding assays indicated that NFe is different from both AP-1 and ATF.
机译：已发现大鼠胚胎肌球蛋白重链启动子（MHCemb）的DNA片段特异性结合从小鼠C2成肌细胞，成肌细胞和HeLa细胞制备的提取物中存在的核因子（NFe）。结合位点（BSe）的核苷酸序列已被鉴定为5'-GTGTCAGTCA-3'，位于-93和-84之间。对C2成肌细胞和C2成肌细胞中MHCemb启动子缺失构建体的瞬时表达研究表明，NFe是转录因子。 NFe结合位点的缺失分别导致C2肌管和C2成肌细胞中启动子活性降低4-6倍和2倍。此外，BSe上的点突变不仅消除了MHCemb启动子的NFe结合活性，而且导致启动子活性降低到与C2肌管，成肌细胞和Hela细胞中的缺失构建体相似的水平（四到六倍）。尽管BSe和最近鉴定的转录因子AP-1和ATF的结合位点具有显着的同源性，但是竞争结合试验的结果表明NFe与AP-1和ATF均不同。

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