Regulation of the KCNJ5 gene by SF-1 in the adrenal cortex: Complete genomic organization and promoter function
Regulation of the KCNJ5 gene by SF-1 in the adrenal cortex: Complete genomic organization and promoter function
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DOI：
10.1016/j.mce.2019.110657
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发表时间：
2020-02-05
影响因子：
4.1
通讯作者：
Yamada, Masanobu
中科院分区：
文献类型：
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作者：
Nishikido, Ayaka;Okamura, Takashi;Yamada, Masanobu
KCNJ 5基因的激活突变是导致大量醛固酮腺瘤的原因。为了阐明KCNJ 5表达的分子机制，我们鉴定了整个人KCNJ 5基因。该基因全长约29.8kb，包含3个外显子和2个内含子。KCNJ 5 mRNA在肾上腺中表达最强。启动子区在-1782bp处含有SF-1的推定结合位点。含有-2444 bp的启动子区域的构建体在肾上腺H295 R细胞中表现出最强的启动子活性，并且在SF-1结合位点中引入突变几乎完全废除了启动子活性。此外，缺失突变，EMSA和敲低分析表明，SF-1结合到这个元件，是功能性的。免疫组化显示KCNJ 5主要表达于肾小球，而SF-1则广泛表达于肾上腺皮质。这些结果表明SF-1介导人KCNJ 5在肾上腺皮质中的表达。
Activating mutations in the KCNJ5 gene are responsible for the significant number of aldosterone-producing adenomas. To elucidate the molecular mechanisms underlying KCNJ5 expression, we characterized the entire human KCNJ5 gene. The gene spanned approximately 29.8 kb and contained three exons and two introns. The strongest expression of KCNJ5 mRNA was observed in the adrenal gland. The promoter region contained a putative binding site for SF-1 at -1782 bp. A construct containing - 2444 bp of the promoter region exhibited the strongest promoter activity in adrenal H295R cells, and the introduction of a mutation in the SF-1 binding site almost completely abolished promoter activity. Furthermore, deletion mutation, EMSA, and knockdown analyses revealed that SF-1 bound to this element and was functional. Immunochemistry showed that KCNJ5 was predominantly expressed in the zona glomerulosa, while SF-1 was ubiquitously expressed in the adrenal cortex. These results demonstrated that SF-1 mediates the expression of human KCNJ5 in the adrenal cortex.