Alpha 1-antichymotrypsin/SerpinA3 is a novel target of orphan nuclear receptor Nur77
Alpha 1-antichymotrypsin/SerpinA3 is a novel target of orphan nuclear receptor Nur77
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DOI：
10.1111/j.1742-4658.2008.06269.x
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发表时间：
2008-03-01
期刊：
影响因子：
5.4
通讯作者：
Zheng, Dexian
中科院分区：
文献类型：
--
作者：
Zhao, Yongjuan;Liu, Yanxin;Zheng, Dexian
Nur 77是核受体超家族成员之一。作为一种转录因子，Nur 77参与多种生物学过程，包括T细胞发育、炎症反应、类固醇激素合成和肝脏葡萄糖代谢。它通常通过结合其靶基因启动子区的Nur 77应答元件（NBRE）发挥作用。在本研究中，我们确定了一个新的Nur 77调节基因，α 1-抗糜蛋白酶/SerpinA 3，通过计算预测和湿实验室验证相结合的方法。首先，我们通过人类全基因组扫描确定了483个候选基因的近端启动子中的NBREs。在HEK 293 T细胞中的荧光素酶报告基因测定中，14个功能相关基因中的3个被进一步鉴定为被Nur 77反式激活。反式激活试验证明SerpinA 3启动子区的NBRE（-182到-175）是HEK 293 T和HepG 2细胞中Nur 77依赖的新功能基序。电泳迁移率变化和染色质免疫沉淀分析表明，Nur 77物理协会与SerpinA 3启动子区在体外和体内。Nur 77过表达和RNA干扰介导的Nur 77基因敲低分析证实SerpinA 3确实是一个新的Nur 77靶向基因。这些数据可能揭示Nur 77在炎症反应和急性期反应以及阿尔茨海默病发展中的作用。
Nur77 is one member of the nuclear receptor superfamily. As a transcription factor, Nur77 participates in a variety of biological processes, including T cell development, inflammatory responses, steroid hormone synthesis, and hepatic glucose metabolism. It typically acts via binding to the Nur77 responsive element (NBRE) in the promoter regions of its target genes. In the present study, we identified a novel Nur77-regulated gene, alpha 1-antichymotrypsin/SerpinA3, via an approach combining computational prediction and wet-laboratory validations. First, we identified 483 candidate genes via a human genome-wide scan for NBREs in their proximal promoters. Three out of 14 function-associated genes were further identified to be transactivated by Nur77 in luciferase reporter gene assays in HEK 293T cells. The transactivation assay proved that the NBRE (-182 to -175) in the SerpinA3 promoter region is a novel Nur77-dependent functional motif in HEK 293T and HepG2 cells. Electrophoretic mobility shift and chromatin immunoprecipitation assays demonstrated that Nur77 physically associates with the SerpinA3 promoter region both in vitro and in vivo. Nur77 overexpression and RNA interference-mediated Nur77 gene knockdown analysis confirmed that SerpinA3 is indeed a novel Nur77-targeted gene. These data may throw light on the function of Nur77 in inflammatory responses and acute-phase reactions as well as the development of Alzheimer's disease.