A shared promoter element regulates the expression of three steroidogenic enzymes.
A shared promoter element regulates the expression of three steroidogenic enzymes.
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DOI：
10.1210/mend-5-10-1552
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发表时间：
1991-10
影响因子：
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通讯作者：
D. A. Rice;Andréa;-R.;Mouw;A. M. Bogerd;Keith;-L.;Parker
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文献类型：
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作者：
D. A. Rice;Andréa;-R.;Mouw;A. M. Bogerd;Keith;-L.;Parker
小鼠肾上腺皮质协调表达皮质类固醇生物合成所需的三种细胞色素P450酶：胆固醇侧链裂解酶（SCC）、类固醇21-羟化酶（21-OHase）和类固醇11 β-羟化酶（11 β-OHase）。在它们的5 '-侧翼区域，我们先前鉴定了六个含有AGGTC基序变异的元件，这些基序调节小鼠Y1肾上腺皮质细胞中的表达：-210、-140和-65处的21-OHase元件;-70和-40处的SCC元件;以及-310处的11 β-OHase元件。我们在这里证明，所有六个元素与相同的，或密切相关的，DNA结合蛋白（S）相互作用。首先，这些元素在凝胶迁移试验中均形成具有相似迁移率的复合物，表明它们与相似大小的蛋白质相互作用。用含有精确AGGTCA序列的那些探针观察到额外的较大复合物。第二，竞争实验证实，与不同元素相互作用的因子具有密切相关或相同的识别特异性。最后，在连续色谱柱上分离核蛋白时，观察到难以区分的移位活动。总的来说，这些结果表明，相关元素与一个共享的蛋白质相互作用，以调节三个必需的类固醇生成酶。AGGTCA序列基序包含核激素受体家族几个成员的应答元件。寡核苷酸竞争和特异性的抗血清在凝胶迁移试验中的影响牵连鸡卵清蛋白上游启动子转录因子的形成较大的复合物中看到的元素含有确切的AGGTCA序列。因此，这个核激素受体家族的成员也可以调节肾上腺类固醇生成酶的表达。
The adrenal cortex of the mouse coordinately expresses three cytochrome P450 enzymes that are required for the biosynthesis of corticosteroids: cholesterol side-chain cleavage enzyme (SCC), steroid 21-hydroxylase (21-OHase), and steroid 11 beta-hydroxylase (11 beta-OHase). Within their 5'-flanking regions, we previously identified six elements containing variations of an AGGTC motif that regulated expression in mouse Y1 adrenocortical cells: 21-OHase elements at -210, -140, and -65; SCC elements at -70 and -40; and an 11 beta-OHase element at -310. We demonstrate here that all six elements interact with the same, or closely related, DNA-binding protein(s). First, these elements all formed complexes of similar mobility in gel shift assays, suggesting that they interacted with protein(s) of similar size. Additional larger complexes were seen with those probes containing exact AGGTCA sequences. Second, competition experiments confirmed that the factor(s) interacting with different elements had closely related or identical recognition specificities. Finally, indistinguishable profiles of shift activities were seen upon fractionation of nuclear proteins over sequential chromatographic columns. Collectively, these results suggest that related elements interact with a shared protein to regulate three essential steroidogenic enzymes. An AGGTCA sequence motif comprises the response element for several members of the nuclear hormone receptor family. Oligonucleotide competitions and specific effects of antisera in gel shift assays implicated chicken ovalbumin upstream promoter-transcription factor in the formation of the larger complexes seen with the elements containing exact AGGTCA sequences. Therefore, this member of the nuclear hormone receptor family also may regulate the expression of the adrenal steroidogenic enzymes.