Rosenthal N, Berglund E B, Wentworth B M, Donoghue M, Winter B, Bober E, Braun T, Arnold H H
Department of Biochemistry, Boston University School of Medicine, MA 02118.
Nucleic Acids Res. 1990 Nov 11;18(21):6239-46. doi: 10.1093/nar/18.21.6239.
A potent muscle-specific enhancer element, originally described in the rat myosin light chain (MLC) 1/3 locus located downstream of the coding region, is found in an analogous position in the human MLC1/3 gene. When linked to a CAT reporter gene and transfected into muscle or non-muscle cells, the human MLC enhancer directs high levels of muscle-specific gene expression from homologous or heterologous promoters, irrespective of position or orientation relative to the CAT transcription unit. A significant degree of sequence homology (over 85%) in the 3'-flanking regions of the two MLC genes is restricted to a 200 bp sequence which lies approximately 1.5 kb downstream of the polyadenylation site in both species. The human enhancer sequence includes binding sites for human myogenic determination factors containing a common basic helix-loop-helix motif, and it can be trans-activated to varying degrees in non-muscle cells by these factors. This study establishes the MLC enhancer as an evolutionarily conserved, integral component of the MLC1/3 locus which constitutes a novel target for the action of myogenic determination factors.
一种强效的肌肉特异性增强子元件,最初是在大鼠肌球蛋白轻链(MLC)1/3基因座的编码区下游被发现的,在人类MLC1/3基因的类似位置也能找到。当与CAT报告基因连接并转染到肌肉或非肌肉细胞中时,人类MLC增强子可指导同源或异源启动子高水平的肌肉特异性基因表达,而与相对于CAT转录单元的位置或方向无关。两个MLC基因3'侧翼区域存在显著程度的序列同源性(超过85%),这种同源性局限于一个200 bp的序列,该序列在两个物种中均位于多聚腺苷酸化位点下游约1.5 kb处。人类增强子序列包含含有常见碱性螺旋-环-螺旋基序的人类生肌决定因子的结合位点,并且在非肌肉细胞中它可被这些因子不同程度地反式激活。本研究确定MLC增强子是MLC1/3基因座中一个进化上保守的、不可或缺的组成部分,它构成了生肌决定因子作用的一个新靶点。