INTERACTION OF THE ESTROGEN RECEPTOR WITH DNA

雌激素受体（ER）是一种配体活化蛋白， 雌激素和抗雌激素在靶细胞中的作用。 儿 与靶基因中的雌激素反应元件（ERE）的相互作用是 负责雌激素在调节 乳腺癌细胞和乳腺癌细胞的增殖和其他特性 生殖道 尽管ER与ERE的相互作用 在激活雌激素敏感的 基因，这种相互作用引发基因变化的机制 表达不太清楚。 我们将使用蛋白酶敏感性测定来检查ERE的作用 ER构象的序列，并确定是否雌激素或抗雌激素 结合改变了ERE结合受体的构象。 DNA酶I， 甲基化干扰和羟基自由基足迹将提供 蛋白质-DNA接触的互补信息， ER与共有卵黄蛋白原A2 ERE或 不完全pS2 ERE。 我们将确定DNA诱导的ER变化是否 构象负责ERE结合受体的缔合 以及这些蛋白质的结合是否可以 在雌激素的调节中发挥额外的控制水平- 反应基因 与A2和pS2相关的转录因子 ERE将是孤立的，而那些与 将鉴定A2或pS2 ERE结合的ER。 这些研究将 为雌激素对基因转录的调节提供了新的见解， 更好地了解ER在乳腺癌中的功能， 正常的生殖靶细胞

The estrogen receptor (ER) is a ligand-activated protein that mediates the actions of estrogens and antiestrogens in target cells. ER interaction with estrogen response elements (EREs) in target genes is responsible for the diverse effects of estrogens in regulating proliferation and other properties of breast cancer cells and cells of the reproductive tract. Although the interaction of the ER with the ERE is of paramount importance in the activation of estrogen-responsive genes, the mechanism by which this interaction initiates changes in gene expression is not well understood. We will use protease sensitivity assays to examine the effects of ERE sequence on ER conformation and determine if estrogen or antiestrogen binding alters the conformation of the ERE-bound receptor. DNase I, methylation interference, and hydroxyradical footprinting will provide complementary information about protein-DNA contacts involved in the interaction of ER with the consensus vitellogenin A2 ERE or the imperfect pS2 ERE. We will determine if DNA-induced changes in ER conformation are responsible for association of the ERE-bound receptor with selected proteins and whether the binding of these proteins could exert an additional level of control in the regulation of estrogen- responsive genes. Transcription factors associated with the A2 and pS2 EREs will be isolated and those which uniquely associate with either the A2 or the pS2 ERE-bound ER will be identified. These studies will provide new insights into estrogen regulation of gene transcription and a better understanding of how the ER functions in breast cancer and normal reproductive target cells.