Investigation on the transcription factors of porcine 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase genes
Investigation on the transcription factors of porcine 3β-hydroxysteroid dehydrogenase and 17β-hydroxysteroid dehydrogenase genes
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猪3β-羟基类固醇脱氢酶和17β-羟基类固醇脱氢酶基因转录因子的调查。
DOI：
10.1016/j.gene.2012.02.039
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发表时间：
2012-05-10
期刊：
影响因子：
3.5
通讯作者：
Fang, Meiying
中科院分区：
文献类型：
--
作者：
Dong, Xinxing;Bai, Ying;Fang, Meiying
3β - 羟类固醇脱氢酶（3β - HSD）和17β - 羟类固醇脱氢酶（17β - HSD）调节哺乳动物的类固醇代谢。在本研究中，我们旨在对这两个基因5'侧翼区域的类固醇相关转录因子进行特性分析。将这两个基因上约1kb的5'侧翼区域进行一系列5'缺失，然后与含有萤火虫荧光素酶cDNA的pGL3基本载体融合，再转染到人肝细胞肝癌细胞系（HepG2）中。荧光素酶活性测定表明，3β - HSD1启动子的 - 574至 - 617bp区域以及17β - HSD7启动子的 - 850至 - 868bp区域诱导出最高的荧光素酶活性。通过序列同源性分别鉴定出一种假定的转录因子，即3β - HSD1基因的富含脯氨酸和酸性氨基酸的碱性亮氨酸拉链（PAR / bZIP）家族，以及17β - HSD7的三氨基酸环延伸（TALE）同源结构域类别。凝胶迁移实验进一步证实了假定元件与核提取物的结合能力。我们的研究为3β - HSD1和17β - HSD7的转录调控提供了新的见解，并进一步暗示了它们在类固醇代谢中的作用。（C）2012爱思唯尔有限公司。保留所有权利。
The enzymes 3 beta-hydroxysteroid dehydrogenase (3 beta HSD) and 17 beta-hydroxysteroid dehydrogenase (17 beta HSD) regulate the steroid metabolism in mammals. In this study, we aimed to characterize the steroid related transcription factors at the 5' flanking region of these two genes. A series of 5' deletions of approximately 1 kb of 5'-flanking region on both genes were fused to a pGL3 basic vector containing firefly luciferase cDNA, and then transfected to human hepatocellular liver carcinoma cell line (HepG2). Luciferase activity assay indicated the region from -574 to -617 bp of the 3 beta HSD1 promoter, and from -850 to -868 bp of 17 beta HSD7 promoter induced the highest luciferase activity. A putative transcription factor, i.e. the proline and acidic amino acid-rich basic leucine zipper (PAR/bZIP) family of 3 beta HSD1 gene, and three-amino acid loop extension (TALE) homeodomain class of 17 beta HSD7 were identified respectively by sequence homology. Gel shift assay further confirmed the binding capacity of the putative elements to nuclear extract. Our study gives new insights to the transcriptional regulation of 3 beta HSD1 and 17 beta HSD7 and further hints to their involvement in steroid metabolism. (C) 2012 Elsevier B.V. All rights reserved.