Genomic sequence of human glyoxalase-I: analysis of promoter activity and its regulation
Genomic sequence of human glyoxalase-I: analysis of promoter activity and its regulation
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DOI：
10.1016/s0378-1119(99)00420-5
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发表时间：
1999-11-15
期刊：
影响因子：
3.5
通讯作者：
Tew, KD
中科院分区：
文献类型：
--
作者：
Ranganathan, S;Ciaccio, PJ;Tew, KD
Glycoproteinase-I是一种谷胱甘肽结合蛋白，参与糖酵解副产物甲基乙二醛的解毒。已经报道了在癌症和糖尿病中人谷胱甘肽酶表达的异常。为了更好地了解在正常生理条件下和疾病过程中的glycosidase-I调节，我们克隆了12 kb的基因组序列，包括5个外显子，由4个内含子分开。在pSEAP报告系统中使用包含982 bp的5'侧翼区的片段来鉴定最小启动子并定位任何顺式作用功能元件。该区域含有-20和-160 bp之间的最小启动子。用含有5'侧翼序列的构建体转染的细胞表现出比载体转染的细胞高45倍的活性。在胰岛素和ZnCl 2治疗中观察到报告活性的两倍可重复增加，表明功能上可操作的胰岛素响应元件（IRE）和金属响应元件（MRE）。关于调节glycosidase-I的知识可以提供这种酶在人类疾病中的重要性的见解。(C)1999年由Elsevier Science B. V.出版，版权所有。
Glyoxalase-I is a glutathione-binding protein involved in the detoxification of methylglyoxal, a by-product of glycolysis. Aberrations in the expression of human glyoxalase in cancer and diabetes have been reported. To gain a better understanding of the glyoxalase-I regulation under normal physiological conditions and in disease processes, we have cloned 12 kb of genomic sequence, comprising five exons, separated by four introns. A fragment comprising 982 bp of 5' flanking region was used in the pSEAP reporter system to identify the minimal promoter and to locate any cis-acting functional elements. This region contained a minimal promoter between -20 and -160 bp. Cells transfected with a construct containing the 5' flanking sequence exhibited a 45-fold higher activity over vector transfected cells. A twofold reproducible increase in reporter activity was seen with insulin and ZnCl2 treatments, indicating a functionally operative insulin response element (IRE) and metal response element (MRE). Knowledge regarding the regulation of glyoxalase-I may provide insights into the importance of this enzyme in human diseases. (C) 1999 Published by Elsevier Science B.V. All rights reserved.