A conserved antioxidant response element (ARE) in the promoter of human carbonyl reductase 3 (CBR3) mediates induction by the master redox switch Nrf2.
A conserved antioxidant response element (ARE) in the promoter of human carbonyl reductase 3 (CBR3) mediates induction by the master redox switch Nrf2.
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DOI：
10.1016/j.bcp.2011.09.027
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发表时间：
2012-01-01
影响因子：
5.8
通讯作者：
Blanco JG
中科院分区：
文献类型：
--
作者：
Cheng Q;Kalabus JL;Zhang J;Blanco JG
羰基还原酶活性催化几种内源性和外源性羰基底物的双电子还原。最近的数据表明，人羰基还原酶3 （CBR3）的表达受主氧化还原开关Nrf2的调控。Nrf2与靶基因启动子中的保守抗氧化反应元件（AREs）结合。CBR3启动子中功能性AREs的存在尚未有报道。在本研究中，构建报告基因的实验表明，典型Nrf2激活剂叔丁基对苯二酚（t-BHQ）在HepG2细胞和MCF-7细胞中诱导CBR3启动子活性（2.7倍，p<0.05）和（22倍，p<0.01）。计算搜索在CBR3远端启动子区域发现了一个保守的ARE（−2698ARE）。从4212 bp的CBR3启动子结构中删除该ARE会影响启动子的基础活性和对t-BHQ处理的启动子活性的诱导。−2698ARE的缺失也影响了过表达Nrf2的细胞中CBR3启动子活性的诱导。电泳迁移迁移试验（EMSA）表明，在t-BHQ处理的细胞核提取物中，特定蛋白复合物与−2698ARE的结合增加。用抗Nrf2抗体进行EMSA实验，证实Nrf2存在于特异性核蛋白- 2698ARE复合物中。这些数据表明，远端- 2698ARE介导了Nrf2原型激活因子对人CBR3的诱导。
Carbonyl reductase activity catalyzes the two electron reduction of several endogenous and exogenous carbonyl substrates. Recent data indicate that the expression of human carbonyl reductase 3 (CBR3) is regulated by the master redox switch Nrf2. Nrf2 binds to conserved antioxidant response elements (AREs) in the promoters of target genes. The presence of functional AREs in the CBR3 promoter has not yet been reported. In this study, experiments with reporter constructs showed that the prototypical Nrf2 activator tert-butyl hydroquinone (t-BHQ) induces CBR3 promoter activity in cultures of HepG2 (2.7-fold; p<0.05) and MCF-7 cells (22-fold; p<0.01). Computational searches identified a conserved ARE in the distal CBR3 promoter region (−2698ARE). Deletion of this ARE from a 4212-bp CBR3 promoter construct impacted basal promoter activity and induction of promoter activity in response to treatment with t-BHQ. Deletion of −2698ARE also impacted the induction of CBR3 promoter activity in cells overexpressing Nrf2. Electrophoretic mobility shift assays (EMSA) demonstrated increased binding of specific protein complexes to −2698ARE in nuclear extracts from t-BHQ treated cells. The presence of Nrf2 in the specific nuclear protein-−2698ARE complexes was evidenced in EMSA experiments with anti-Nrf2 antibodies. These data suggest that the distal −2698ARE mediates the induction of human CBR3 in response to prototypical activators of Nrf2.
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影响因子：
3.8
作者：
Niture SK;Kaspar JW;Shen J;Jaiswal AK
通讯作者：
Jaiswal AK
影响因子：
2.6
作者：
Fan, Lu;Goh, Boon-Cher;Lee, Soo-Chin
通讯作者：
Lee, Soo-Chin
DOI：
10.1073/pnas.85.10.3585
发表时间：
1988-05-01
影响因子：
11.1
作者：
OLSON, RD;MUSHLIN, PS;BOUCEK, RJ
通讯作者：
BOUCEK, RJ
影响因子：
7.4
作者：
Cao, Jin;Covarrubias, Vanessa M.;Straubinger, Robert M.;Wang, Hao;Duan, Xiaotao;Yu, Haoying;Qu, Jun;Blanco, Javier G.
通讯作者：
Blanco, Javier G.
影响因子：
20.3
作者：
Macari, Elizabeth R.;Lowrey, Christopher H.
通讯作者：
Lowrey, Christopher H.