Oyhar A, Strangmann-Diekmann M, Kiltz H H, Pongs O
Ruhr-Universität Bochum, Lehrstuhl für Biochemie, Federal Republic of Germany.
Eur J Biochem. 1991 Sep 1;200(2):329-35. doi: 10.1111/j.1432-1033.1991.tb16189.x.
A gel-filtration assay has been developed with which the specific interaction between Drosophila melanogaster ecdysteroid receptor and the 20-hydroxy-ecdysone responsive element of the hsp27 gene promoter region was characterized in terms of complex formation, saturation of DNA binding and the apparent molecular mass of the complex. The hsp27 DNA-binding sequence for ecdysteroid receptor in vitro was delimited by footprinting and mutational analysis. The combined results show that ecdysteroid receptor binds as a dimer to an imperfect palindromic sequence (GGTTCAATGCACT) closely resembling the structures of the different vertebrate steroid-hormone-responsive elements reported so far.
已开发出一种凝胶过滤分析方法,利用该方法从复合物形成、DNA结合饱和度以及复合物的表观分子量等方面对黑腹果蝇蜕皮甾体受体与hsp27基因启动子区域的20-羟基蜕皮酮反应元件之间的特异性相互作用进行了表征。通过足迹法和突变分析确定了体外蜕皮甾体受体的hsp27 DNA结合序列。综合结果表明,蜕皮甾体受体以二聚体形式结合到一个不完全回文序列(GGTTCAATGCACT)上,该序列与迄今为止报道的不同脊椎动物类固醇激素反应元件的结构非常相似。