IDENTIFICATION OF A PUTATIVE ANTIOXIDANT RESPONSE ELEMENT IN THE 5'-FLANKING REGION OF THE HUMAN GAMMA-GLUTAMYLCYSTEINE SYNTHETASE HEAVY SUBUNIT GENE
IDENTIFICATION OF A PUTATIVE ANTIOXIDANT RESPONSE ELEMENT IN THE 5'-FLANKING REGION OF THE HUMAN GAMMA-GLUTAMYLCYSTEINE SYNTHETASE HEAVY SUBUNIT GENE
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DOI：
10.1006/bbrc.1995.1493
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发表时间：
1995-04-06
影响因子：
3.1
通讯作者：
GIPP, JJ
中科院分区：
文献类型：
--
作者：
MULCAHY, RT;GIPP, JJ
我们从P1文库中克隆了人γ-谷氨酰半胱氨酸合成酶重亚基基因（GCS（h）），并从P1克隆的5 ′端分离到一个5.5kb的片段（P1-GCS 5 ′）。P1-GCS 5 '已从-1460到+547测序。通过引物延伸和S1核酸酶保护鉴定多个转录起始位点。通过引物延伸分析在共有TATAAAA框的23 bp（+1和+10）内鉴定两个起始位点;所有序列相对于这两个位点的最5 '编号。在-106和+398处发现了另外两个主要起始位点。后一个位点是所有起始位点中最突出的。除了TATA盒之外，启动子还含有位于TATAAAA上游和下游的-125的CCAAT盒和GC盒。此外，序列的前几百个碱基对是高度富含GC的（类似于75%）。该序列还含有几个Sp-1结合位点、一个共有AP-1位点和几个AP-1样结合位点，以及推定的AP-2位点。在位置+198处鉴定了共有金属响应元件（MRE）。序列分析还在-862至-853处鉴定了推定的核心（5 '-TGACnnnGCA-3'）抗氧化反应元件（ARE）。如其他战神的典型，第二AP-1样序列位于核心序列附近。这些结果表明，GCS（h）基因表达在氧化挑战的反应，可能是通过一个抗氧化反应元件，最近检测到的几个II相酶的启动子区域类似的调节。(The人GCS重亚基基因的5 '侧翼区的序列已被指定登录号L39773）。(C)出版社：Academic Press
We have cloned the human gamma-glutamylcysteine synthetase heavy subunit gene (GCS(h)) from a P1 library and isolated a 5.5kb fragment (P1-GCS5') from the 5'-end of the P1 clone. P1-GCS5' has been sequenced from -1460 to +547. Multiple transcription start sites were identified by primer extension and S1 nuclease protection. Two start sites were identified by primer extension analysis within 23 bp (+1 and +10) of a consensus TATAAAA box; all sequences were numbered relative to the 5'-most of these two sites. Two additional major start sites were identified at -106 and +398. This latter site was the most prominent of all the initiation sites. In addition to a TATA box, the promoter contains a CCAAT box at -125 and GC boxes up- and down-stream of the TATAAAA. In addition, the first few hundred base pairs of the sequence are highly GC-rich(similar to 75%). This sequence also contains several Sp-l binding sites, a consensus AP-1 site and several AP-l-like binding sites, as well as putative AP-2 sites. A consensus metal responsive element (MRE) was identified at position +198. Sequence analysis also identified a putative core (5'-TGACnnnGCA-3') antioxidant response element (ARE) at -862 to -853. As is typical of other AREs, a second AP-l-like sequence is located adjacent to the core sequence. These results suggest that GCS(h) gene expression in response to oxidative challenge may be regulated through an antioxidant response element similar to those recently detected in the promoter region of several Phase II enzymes. (The sequence of the 5'-flanking region of the human GCS heavy subunit gene has been assigned accession number L39773.) (C) 1995 Academic Press, Inc.