THE MITOCHONDRION AS A PRIMARY SITE OF ACTION OF GLUCOCORTICOIDS - THE INTERACTION OF THE GLUCOCORTICOID RECEPTOR WITH MITOCHONDRIAL-DNA SEQUENCES SHOWING PARTIAL SIMILARITY TO THE NUCLEAR GLUCOCORTICOID-RESPONSIVE ELEMENTS
THE MITOCHONDRION AS A PRIMARY SITE OF ACTION OF GLUCOCORTICOIDS - THE INTERACTION OF THE GLUCOCORTICOID RECEPTOR WITH MITOCHONDRIAL-DNA SEQUENCES SHOWING PARTIAL SIMILARITY TO THE NUCLEAR GLUCOCORTICOID-RESPONSIVE ELEMENTS
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DOI：
10.1016/0960-0760(95)00159-w
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发表时间：
1995-10-01
影响因子：
4.1
通讯作者：
SEKERIS, CE
中科院分区：
文献类型：
--
作者：
DEMONACOS, C;DJORDJEVICMARKOVIC, R;SEKERIS, CE
研究了与糖皮质激素反应元件一致序列（GRE）高度相似的6个线粒体基因组序列，其中4个位于细胞色素c氧化酶（COX）亚基I和III基因（GREs I- iv）， 2个位于d环区（GREs a和b），作为大鼠肝细胞质中糖皮质激素受体（GR）的结合位点。从大鼠肝细胞质中纯化的GR与所有潜在的线粒体GREs具有高特异性结合，如过滤器保留和凝胶移位试验所示。地塞米松诱导小鼠线粒体提取物中存在的蛋白质与所有六种假定的线粒体GREs的特异性结合也用相同的方法记录。在凝胶阻滞实验中，纯化的GR和线粒体提取物中的蛋白质都具有相同的条带。利用单特异性抗糖皮质激素受体多克隆抗体（EP），获得了凝胶阻滞蛋白- dna带的超移。这些结果表明，所检查的线粒体基因组序列具有GREs的特征，因为它们显示出特异性结合各自受体蛋白的能力。这些发现支持了线粒体基因组是类固醇和甲状腺激素主要作用位点的假设（Sekeris c.e.：线粒体基因组：类固醇激素可能的主要作用位点，In vivo 4(1990) 317-320）。
Six mitochondrial genome sequences, showing strong similarity to the glucocorticoid responsive element consensus sequence (GRE), four localized within the cytochrome c oxidase (COX) subunit I and III genes (GREs I-IV) and two within the D-loop region (GREs a and b) have been examined as binding sites of glucocorticoid receptor (GR) from rat liver cytosol. Purified GR from rat liver cytosol binds with high specificity to all potential mitochondrial GREs, as shown by filter retention and gel shift assays. Specific binding of protein(s), present in a mitochondrial extract from dexamethasone-induced mice, to all six putative mitochondrial GREs was also documented by the same methodology. Both purified GR and protein(s) from mitochondrial extract give the same band in the gel retardation assay. Using monospecific anti-glucocorticoid receptor polyclonal antibody (EP), a supershift of the gel retarded protein-DNA band was obtained. These results demonstrate that the mitochondrial genome sequences examined have characteristics of GREs, since they show the capacity to specifically bind the respective receptor protein. These findings support the hypothesis that the mitochondrial genome is a primary site of action of steroid and thyroid hormones (Sekeris C.E.: The mitochondrial genome: a possible primary site of action of steroid hormones, In vivo 4 (1990) 317-320).