CHARACTERIZATION OF THE HORMONE RESPONSIVE ELEMENT INVOLVED IN THE REGULATION OF THE PROGESTERONE-RECEPTOR GENE
CHARACTERIZATION OF THE HORMONE RESPONSIVE ELEMENT INVOLVED IN THE REGULATION OF THE PROGESTERONE-RECEPTOR GENE
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DOI：
10.1002/j.1460-2075.1991.tb07713.x
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发表时间：
1991-07-01
期刊：
影响因子：
11.4
通讯作者：
MILGROM, E
中科院分区：
文献类型：
--
作者：
SAVOURET, JF;BAILLY, A;MILGROM, E
孕激素受体基因的转录由雌激素诱导，并由孕激素降低。进行研究以确定基因的区域和所涉及的分子机制。在异源基因前使用该基因的5'侧翼区直到-2762，不能观察到激素调节。只有当使用延伸到+788的基因片段时，才能观察到雌激素和孕酮的调节。从5'和3'端进行性缺失、定点突变和用纯化的雌激素受体进行的DNA酶保护实验表明，具有生物活性的雌激素反应元件（ERE）存在于+698/+723处，与翻译起始重叠。合成了一种带有这种ERE的寡核苷酸，并显示出对异源基因的雌激素诱导作用。它的调节抗雌激素对应的原位孕酮受体基因，因为他莫昔芬是一个部分激动剂，而ICI 164384是一个完整的拮抗剂。在孕酮受体存在的情况下，该ERE也介导孕激素的下调，即使它没有孕酮受体结合能力。DNA酶足迹法表明，这种影响是不是由于雌激素受体的雌激素受体亲和力的ERE在孕激素受体的存在下，减少。最后，使用孕酮受体的缺失突变体表明，类固醇结合和DNA结合结构域是下调所必需的，而N-末端结构域的各个部分的缺失没有影响。
The transcription of the progesterone receptor gene is induced by estrogens and decreased by progestins. Studies were performed to define the regions of the gene and the molecular mechanisms involved. No hormonal regulation could be observed using 5' flanking regions of the gene up to -2762 in front of a heterologous gene. Estrogen and progestin regulation could be observed only when using fragments of the gene extending down to +788. Progressive deletions from the 5' and 3' ends, site-directed mutagenesis and DNase protection experiments with purified estrogen receptor suggested that the biologically active estrogen responsive element (ERE) is present at +698/+723, overlapping the initiation of translation. An oligonucleotide was synthesized bearing this ERE and shown to impart estrogen inducibility to a heterologous gene. Its regulation by anti-estrogens corresponded to that of the in situ progesterone receptor gene since tamoxifen was a partial agonist whereas ICI 164384 was a full antagonist. This ERE also mediated down-regulation by progestins in the presence of the progesterone receptor, even though it has no progesterone receptor binding ability. DNase footprinting showed that this effect was not due to a decrease of estrogen receptor affinity for the ERE in the presence of progesterone receptor. Finally, use of deletion mutants of the progesterone receptor showed that the steroid binding and the DNA binding domains were necessary for down-regulation whereas deletions of various parts of the N-terminal domain were without effect.