Steroidogenic factor-1 binding and transcriptional activity of the cholesterol side-chain cleavage promoter in rat granulosa cells.
Steroidogenic factor-1 binding and transcriptional activity of the cholesterol side-chain cleavage promoter in rat granulosa cells.
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DOI：
10.1210/endo.134.3.8119192
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发表时间：
1994-03
期刊：
影响因子：
4.8
通讯作者：
J. W. Clemens;D. Lala;K. L. Parker;J. S. Richards
中科院分区：
文献类型：
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作者：
J. W. Clemens;D. Lala;K. L. Parker;J. S. Richards
胆固醇侧链裂解细胞色素P450（CYP 11 A; P450 SCC）基因在大鼠卵泡中表达，对促性腺激素（FSH/LH）和cAMP产生应答。为了鉴定大鼠P450 SCC启动子内的功能区域，将5 '-侧翼序列的894个碱基对（bp）和5'-缺失（在-379、-101、-73和-38 bp处）与人GH报告基因连接，并转染到培养的大鼠颗粒细胞中。大鼠启动子的cAMP诱导定位于大鼠启动子内含有两个AGGT/CC/TA基序之一的区域（-73/-38 bp之间），所述基序被命名为SCC 1（-51/-43 bp）和SCC 2（-79/-71 bp）。颗粒细胞中与SCC 1结合的一种核蛋白被鉴定为孤儿受体，类固醇生成因子-1（SF-1）。相反，SCC 2形成了多种蛋白质-DNA复合物，其中只有一种被明确鉴定为SF-1。核提取物结合是序列特异性的，SCC 1结合SF-1比SCC 2更强。因此，大鼠启动子的两个AGGT/CC/TA基序在结构和功能上似乎不同。此外，由于SF-1 mRNA的表达先于P450 SCC mRNA的激素/cAMP诱导，并且在体外不受cAMP调节，因此SF-1在P450 SCC基因转录调节中的功能作用，包括其受cAMP诱导的功能作用，并不完全清楚，可能依赖于其他因素和/或修饰（磷酸化？）SF-1
The cholesterol side-chain cleavage cytochrome P450 (CYP11A; P450scc) gene is expressed in rat ovarian follicles in response to gonadotropins (FSH/LH) and cAMP. To identify functional regions within the rat P450scc promoter, 894 basepairs (bp) of 5'-flanking sequence and 5'-deletions (at -379, -101, -73, and -38 bp) were linked to the human GH reporter gene and transfected into cultured rat granulosa cells. cAMP inducibility of the rat promoter was localized to a region (between -73/-38 bp) that contains one of two AGGT/CC/TA motifs, designated SCC1 (-51/-43 bp) and SCC2 (-79/-71 bp), within the rat promoter. One of the nuclear proteins in granulosa cells that binds to SCC1 was identified as the orphan receptor, steroidogenic factor-1 (SF-1). In contrast, multiple protein-DNA complexes formed with SCC2, only one of which was clearly identified as SF-1. Nuclear extract binding was sequence specific; SCC1 bound SF-1 more strongly than did SCC2. Thus, the two AGGT/CC/TA motifs of the rat promoter appear to differ structurally and functionally. Furthermore, because the expression of SF-1 mRNA precedes hormonal/cAMP induction of P450scc mRNA and is not regulated in vitro by cAMP, the functional role of SF-1 in transcriptional regulation of the P450scc gene, including its induction by cAMP, is not entirely clear and is probably dependent on other factors and/or the modification (phosphorylation?) of SF-1.