Signal transducer and activator of transcription 6 directly regulates human ORMDL3 expression
Signal transducer and activator of transcription 6 directly regulates human ORMDL3 expression
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DOI：
10.1111/febs.12225
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发表时间：
2013-05
期刊：
影响因子：
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通讯作者：
R. Qiu;Yang Yang-Yang;Hailing Zhao;Jiangxia Li;Qian Xin;Shan Shan-Shan;Yongchao Liu;J. Dang;Xiao Yu-Xiao-Y
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文献类型：
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作者：
R. Qiu;Yang Yang-Yang;Hailing Zhao;Jiangxia Li;Qian Xin;Shan Shan-Shan;Yongchao Liu;J. Dang;Xiao Yu-Xiao-Y
OroSomucid-like 3(ORMDL3)与哮喘和一系列自身免疫性疾病有关，并参与内质网介导的炎症反应。然而，它的临床意义和表达的分子机制仍很不清楚。为了阐明人类ORMDL3转录调控的机制，我们克隆了一段1.5kb的基因组DNA片段，该片段含有可能的启动子区域，并通过缺失分析在荧光素酶报告系统中评价了其转录活性。我们确定了一个68bp的区域，其功能是一个最小的启动子。生物信息学分析预测，−至−56bp区域含有信号转导和转录激活因子6(STAT6)结合位点。凝胶迁移率改变分析和染色质免疫沉淀表明，STAT6结合在ORMDL3启动子内的结合部位。包含STAT6结合基序的ORMDL3启动子分别过表达或下调反式激活或反式抑制。IL-4或IL-13处理可增加ORMDL3启动子活性和内源性ORMDL3表达。免疫沉淀和芯片/再芯片分析表明，STAT6和p300存在于与ORMDL3启动子结合的同一蛋白质复合体中。我们的研究证实，STAT6通过直接与启动子区域结合，在调节人ORMDL3的表达中发挥重要作用，这可能有助于揭示STAT6在人类各种疾病中的可能作用。
Orosomucoid‐like 3 (ORMDL3) has been associated with asthma and a series of autoimmune disorders, and is involved in endoplasmic reticulum‐mediated inflammatory responses. However, its clinical significance and the molecular mechanism underlying its expression are still largely unclear. To elucidate the mechanisms of human ORMDL3 transcriptional regulation, we cloned a 1.5 kb genomic DNA fragment containing the putative promoter region and evaluated its transcriptional activity in a luciferase reporter system by deletion analysis. We identified a 68 bp region that functions as a minimal promoter. Bioinformatics analysis predicted that the −64 to −56 bp region contained a signal transducer and activator of transcription 6 (STAT6) binding site. Electrophoretic mobility shift assay and chromatin immunoprecipitation demonstrated that STAT6 bound to its binding site within the ORMDL3 promoter. STAT6 over‐expression or knockdown trans‐activated or trans‐inhibited, respectively, the ORMDL3 promoter containing the STAT6‐binding motif. Treatment with interleukins 4 or 13 increased ORMDL3 promoter activity as well as endogenous ORMDL3 expression. Immunoprecipitation and ChIP/Re‐ChIP assays revealed that STAT6 and p300 exist in the same protein complex that binds to the ORMDL3 promoter. Our study confirmed that STAT6 plays important roles in regulating the expression of human ORMDL3 by directly binding to the promoter region, which may shed light on a possible role in various human diseases.