Regulation of mouse GADD34 gene transcription after DNA damaging agent methylmethane sulfonate
Regulation of mouse GADD34 gene transcription after DNA damaging agent methylmethane sulfonate
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DOI：
10.1016/j.gene.2004.04.008
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发表时间：
2004-07-07
期刊：
影响因子：
3.5
通讯作者：
Isobe, K
中科院分区：
文献类型：
--
作者：
Haneda, M;Xiao, HY;Isobe, K
GADD 34基因是由生长停滞和DNA损伤引起的转录诱导。然而，转录调控的机制仍然不清楚。我们分析了小鼠GADD 34基因的启动子以及甲基甲烷磺酸盐（MMS）诱导的该基因的转录调节。通过引入基因组突变体，这是连接到荧光素酶报告，到NIH 3 T3细胞，我们定义了一个100 bp的片段上游的转录起始位点的GADD 34基因的最小启动子。随后的研究表明，位于这个最小启动子的CRE结合位点是MMS诱导GADD 34基因转录的关键。体外结合实验表明，CRE/DNA复合物中含有磷酸化的c-Jun。c-Jun的显性负性形式的过表达导致MMS响应性启动子活性的降低。从这些结果中，我们得出结论，GADD 34启动子的CRE位点对于MMS响应性顺式元件是不可或缺的，c-Jun是MMS刺激调节GADD 34基因表达的必需转录因子，并且上游信号传导依赖于JNK。(C)2004 Elsevier B. V.保留所有权利。
The GADD34 gene is transcriptionally induced by growth arrest and DNA damage. However, the mechanisms underlying the transcriptional regulation are still unclear. We analyzed the promoter of mouse GADD34 gene and the methylmethane sulfonate (MMS)-induced transcriptional regulation of this gene. By introducing genome mutants, which were linked to the luciferase reporter, into NIH3T3 cells, we defined a 100-bp fragment upstream of the transcriptional initiating site as the minimal promoter of the GADD34 gene. Subsequent study revealed that CRE-binding site located in this minimal promoter was critical for MMS-induced transcription of the GADD34 gene. In vitro binding experiments showed that phosphorylated c-Jun was contained in the CRE/DNA complex. Overexpression of the dominant negative form of c-Jun led to a decrease of MMS-responsive promoter activity. From these results, we conclude that the CRE site of the GADD34 promoter is indispensable to the MMS-responsive cis-element that c-Jun is the essential transcription factor for MMS-stimulated regulation of GADD34 gene expression and that the upstream signaling is dependent on JNK. (C) 2004 Elsevier B.V. All rights reserved.