Quantitative analysis of the glucocorticoid receptor-DNA interaction at the mouse mammary tumor virus glucocorticoid response element.
Quantitative analysis of the glucocorticoid receptor-DNA interaction at the mouse mammary tumor virus glucocorticoid response element.
复制标题
小鼠乳腺肿瘤病毒糖皮质激素反应元件的糖皮质激素受体-DNA 相互作用的定量分析。
DOI：
10.1016/s0021-9258(17)44892-7
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发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Örjan Wrange
中科院分区：
文献类型：
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作者：
Thomas Perlmann;Per Eriksson;Örjan Wrange
用纯化的大鼠肝糖皮质激素受体（GR）定量分析了小鼠乳腺肿瘤病毒5 ′-末端重复序列中GR特异性结合片段的蛋白质-DNA相互作用。产生了截短的受体，并用于证明异二聚体GR的形成，其进一步显示与受体单体快速平衡。与随机小牛胸腺DNA相比，GR与特定GR位点结合的相对亲和力约为2 x 10（3）。在平衡状态下，小鼠乳腺肿瘤病毒5 '-长末端重复序列中两个功能重要的DNA位点达到半饱和需要3 × 10（-10）M的游离GR浓度。虽然这两个DNA片段协同作用，介导激素反应，我们没有检测到合作GR结合这些地区在体外。然而，GR在下游结合区域内协同结合。类似地，GR不能促进因子结合到邻近的核因子1位点，这是启动子中的另一个必需元件。与此相反，核因子1结合略有抑制GR。
Purified glucocorticoid receptor (GR) from rat liver was used for a quantitative analysis of the protein-DNA interaction at specific GR-binding segments within the 5‘-long terminal repeat of the mouse mammary tumor virus. A truncated receptor was generated and used to demonstrate formation of heterodimeric GR, which furthermore was shown to be in rapid equilibrium with receptor-monomer. The relative affinity for GR binding to specific GR sites versus random calf thymus DNA was approximately 2 x 10(3). At equilibrium a free GR concentration of 3 x 10(-10) M was required for half-maximal saturation of the two functionally important DNA sites within the mouse mammary tumor virus 5‘-long terminal repeat. Although these two DNA segments act synergistically in mediating hormonal response, we did not detect cooperative GR binding to these regions in vitro. However, GR bound cooperatively within the downstream binding region. Similarly, GR was unable to facilitate factor binding to a neighboring nuclear factor 1 site, another essential element in the promoter. In contrast, nuclear factor 1 binding was inhibited slightly by GR.