Walsh K, Schimmel P
J Biol Chem. 1987 Jul 15;262(20):9429-32.
We show that DNA fragments, which span the skeletal muscle alpha-actin promoter, form specific complexes with proteins from nuclear extracts of 11 different cultured cell types. These include two myocyte stage specific cell types, myoblast and myotube. The myocyte nuclear extracts are distinguished from all of the others by the electrophoretic mobility of the predominant DNA complex. This complex is greatly diminished in the nine non-myocyte cell types, where a different complex predominates. The major myocyte and non-myocyte complexes are caused by distinct binding activities that compete for the same DNA sequence element. DNA footprint analysis shows that this element is located 78 nucleotides upstream of the transcription start site, within a region that appears essential for expression in developing myocytes.
我们发现,跨越骨骼肌α-肌动蛋白启动子的DNA片段能与来自11种不同培养细胞类型的核提取物中的蛋白质形成特异性复合物。其中包括两种成肌细胞阶段特异性细胞类型,即成肌细胞和肌管。通过主要DNA复合物的电泳迁移率可将成肌细胞核提取物与其他所有提取物区分开来。在9种非成肌细胞类型中,这种复合物显著减少,在这些细胞中另一种复合物占主导地位。主要的成肌细胞和非成肌细胞复合物是由竞争相同DNA序列元件的不同结合活性引起的。DNA足迹分析表明,该元件位于转录起始位点上游78个核苷酸处,处于一个对发育中的成肌细胞表达似乎至关重要的区域内。