Two adjacent cis-regulatory elements are required for ecdysone response of ecdysone receptor (EcR) B1 transcription.
Two adjacent cis-regulatory elements are required for ecdysone response of ecdysone receptor (EcR) B1 transcription.
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ecdysone受体（ECR）B1转录需要两个相邻的顺式调节元件。
DOI：
10.1371/journal.pone.0049348
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Fujiwara H
中科院分区：
文献类型：
--
作者：
Shirai H;Kamimura M;Yamaguchi J;Imanishi S;Kojima T;Fujiwara H
三类不同的核受体，ECR，E75和HR3，是昆虫中蜕皮激素诱导的基因激活级联反应的关键调节因子。这些基因的转录是由蜕皮激素(20E)不同地诱导的，尽管它们对20E的反应的详细机制在很大程度上尚不清楚。我们利用荧光素酶报告基因分析，在对蜕皮激素敏感的家蚕细胞系NIAS-BM-AFF3(AFF3)中，鉴定了家蚕BmEcR-B1、BmE75-A和BHR3-B亚型基因启动子中存在的蜕皮激素反应元件(ECRE)。BmEcR-B1在−2800的ECRE由两个相邻的元件组成，隔开5个核苷酸，E1(15个核苷酸)和E2(21个核苷酸)，这两个元件都是20E反应所必需的。进一步的电泳迁移率改变分析表明，E1与ECR/USP异源二聚体结合，而E2可能与E-box(CACGTG)结合因子如bHLH蛋白结合。在7种鳞翅目昆虫ECR-B1的启动子上游区域也检测到独特的E1+E2型ECRE。BmE75-A启动子中的20bpECRE和BHR3-B启动子中的18bpECRE只含有E1型ECR/USP结合元件，而E2型元件不在这些基因的启动子区域。启动子区域存在的E2元件或其他顺式调节元件的组合解释了每类核受体基因的不同20E反应。此外，ECR-B1的E1+E2结构可能参与蜕皮激素和其他调节途径之间的串扰。
Three distinct classes of nuclear receptors, EcR, E75, and HR3, are key regulators in the ecdysone-inducible gene activation cascade in insects. The transcription of these genes is induced by ecdysone (20E) differently, although the detailed mechanisms underlying their responses to 20E are largely unknown. We identified ecdysone response elements (EcREs) present in the promoters of genes coding BmEcR-B1, BmE75-A, and BHR3-B isoforms from Bombyx mori employing luciferase reporter assays in an ecdysteroid-responsive cultured cell line, NIAS-Bm-aff3 (aff3). The EcRE of BmEcR-B1 at −2800 comprises of two adjacent elements separated by 5 bp, E1 (15 bp) and E2 (21 bp), both of which are required for the 20E response. Further analysis using electrophoretic mobility shift assays showed that E1 binds to the EcR/USP heterodimer and that E2 may bind to the E-box (CACGTG) binding factor such as bHLH protein. The unique E1+E2-type EcRE is also detected in the promoter upstream regions of EcR-B1 from seven lepidopteran species studied. In contrast, both a 20 bp EcRE identified in the promoter of BmE75-A and a 18 bp EcRE identified in the BHR3-B promoter, contained only E1-type EcR/USP binding element but the E2 type element was not in the promoter regions of these genes. The combination of presence of the E2 element or other cis-regulatory elements in promoter regions explains the different 20E response of each class of nuclear receptor genes. Furthermore, the E1+E2 structure for EcR-B1 can be involved in a possible cross-talk between ecdysteroid and other regulatory pathways.
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