Crone D E, Kim H S, Spindler S R
Department of Biochemistry, University of California, Riverside 92521.
J Biol Chem. 1990 Jul 5;265(19):10851-6.
We report that alpha and beta type rat thyroid hormone receptors bind specifically and with high affinity to the 10-base pair sequence immediately 3' of the rat growth hormone TATA box (positions -25 to -16) in a region of the rat growth hormone promoter which can be negatively hormone responsive (nTRE). The receptors have approximately 7-fold lower affinity in vitro for the nTRE than for the thyroid hormone-responsive enhancer of the rat growth hormone gene (TRE). Proteins extracted with high salt concentration from rat pituitary cell nuclei enhance binding of the receptors to both the TRE and nTRE. A modification of the avidin-biotin complex DNA binding assay which enhances the sensitivity of the assay approximately 100-fold was used in these studies. The immediate proximity of a receptor binding site to the rat growth hormone TATA box suggests that direct interaction between receptor and TFIID (the TATA binding protein) mediates nTRE activity.
我们报告称,α型和β型大鼠甲状腺激素受体与大鼠生长激素启动子中位于大鼠生长激素TATA框(位置-25至-16)3'端紧邻的10碱基对序列特异性且高亲和力结合,该区域可呈负性激素反应(nTRE)。在体外,受体对nTRE的亲和力比对大鼠生长激素基因的甲状腺激素反应增强子(TRE)约低7倍。用高盐浓度从大鼠垂体细胞核中提取的蛋白质可增强受体与TRE和nTRE的结合。这些研究中使用了一种改进的抗生物素蛋白-生物素复合物DNA结合测定法,该方法可将测定灵敏度提高约100倍。受体结合位点与大鼠生长激素TATA框紧邻,这表明受体与TFIID(TATA结合蛋白)之间的直接相互作用介导了nTRE活性。