Van Dijck P, Verhoeven G
Laboratorium voor Experimentele Geneeskunde en Endocrinologie, Katholieke Universiteit Leuven, Belgium.
Biochem Biophys Res Commun. 1992 Jan 15;182(1):174-81. doi: 10.1016/s0006-291x(05)80127-5.
Since estrogens strongly suppress the expression of alpha 2u-globulin genes in the rat liver, we studied the binding of estrogen-receptor complexes to fragments derived from alpha 2u-globulin gene RAO 01 using a DNA-cellulose competition assay. Rat uterus cytosol labelled with [3H]estradiol was used as a source of the estrogen receptor. As a positive control in these experiments we used an oligonucleotide containing the estrogen response element (ERE) cloned into pUC18. Our experiments indicate that estrogen-receptor complexes bind specifically to the ERE and to a fragment of RAO 01 located in the 5'-upstream region (bp -606 up to -575). This fragment is conserved among other members of this gene family. This is the first time that in vitro estrogen receptor binding is observed to gene fragments derived from a gene that is repressed by this steroid in vivo.
由于雌激素能强烈抑制大鼠肝脏中α2u-球蛋白基因的表达，我们使用DNA-纤维素竞争分析法研究了雌激素受体复合物与源自α2u-球蛋白基因RAO 01的片段的结合情况。用[3H]雌二醇标记的大鼠子宫胞质溶胶作为雌激素受体的来源。在这些实验中，我们使用克隆到pUC18中的含有雌激素反应元件（ERE）的寡核苷酸作为阳性对照。我们的实验表明，雌激素受体复合物能特异性地结合ERE以及位于5'-上游区域（bp -606至-575）的RAO 01片段。该片段在这个基因家族的其他成员中是保守的。这是首次在体外观察到雌激素受体与源自一个在体内被这种类固醇抑制的基因的基因片段结合。
