THE PROGESTERONE-RECEPTOR STIMULATES CELL-FREE TRANSCRIPTION BY ENHANCING THE FORMATION OF A STABLE PREINITIATION COMPLEX
THE PROGESTERONE-RECEPTOR STIMULATES CELL-FREE TRANSCRIPTION BY ENHANCING THE FORMATION OF A STABLE PREINITIATION COMPLEX
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DOI：
10.1016/0092-8674(90)90740-6
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发表时间：
1990-01-26
期刊：
影响因子：
64.5
通讯作者：
OMALLEY, BW
中科院分区：
文献类型：
--
作者：
KLEINHITPASS, L;TSAI, SY;OMALLEY, BW
高度纯化的鸡孕酮受体（cPR）在体外转录试验中直接刺激RNA合成。通过cPR刺激转录需要在模板中存在孕酮响应元件（PREs），并且可以通过添加含有PREs的竞争性寡核苷酸来特异性抑制。受体与两个PREs的结合是协同的，并且导致转录的协同（27倍）刺激。在大肠杆菌中产生了一种纯化的融合蛋白，其含有与酵母泛素连接的cPR的DNA结合结构域，并且在转录测定中也具有功能。使用这种体外转录系统，我们证明了盐处理激活的无酶cPR以不依赖酶的方式诱导测试基因的转录。最后，我们提出的证据表明，孕激素受体的作用，通过促进在靶基因启动子形成一个稳定的preinitiation复合物，从而增强了启动RNA聚合酶II的转录。
Highly purified chicken progesterone receptor (cPR) is shown to stimulate RNA synthesis directly in an in vitro transcription assay. Stimulation of transcription by cPR requires the presence of progesterone response elements (PREs) in the template and can be specifically inhibited by addition of competitor oligonucleotides containing PREs. Binding of receptor to two PREs is cooperative and leads to a synergistic (27-fold) stimulation of transcription. A purified fusion protein containing the DNA binding domain of cPR linked to yeast ubiquitin was produced in Escherichia coli and also functions in the functions in the transcription assay. Using this in vitro transcription system, we demonstrate that hormone-free cPR activated by salt treatment induces transcription of a test gene in a hormone-independent manner. Finally, we present evidence that the progesterone receptor acts by facilitating the formation of a stable preinitiation complex at the target gene promoter and thus augments the initiation of transcription by RNA polymerase II.