Genomic structure and functional characterization of the human ADAM10 promoter
Genomic structure and functional characterization of the human ADAM10 promoter
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DOI：
10.1096/fj.04-3619fje
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发表时间：
2005-06-01
期刊：
影响因子：
4.8
通讯作者：
Postina, R
中科院分区：
文献类型：
--
作者：
Prinzen, C;Müller, U;Postina, R
ADAM 10基因编码一种膜结合的去整合素-金属蛋白酶，在阿尔茨海默病（AD）小鼠模型中过表达后，可预防淀粉样病变并改善长时程增强和记忆。由于增加ADAM 10的表达似乎是治疗AD的合理方法，我们对ADAM 10基因进行了功能分析。人和小鼠的ADAM 10基因均由16个外显子组成，长度约为160 kbp，在翻译起始位点上游500 bp范围内进化高度保守。通过使用荧光素酶报告基因检测，我们证明，核苷酸-2179到-1上游的人ADAM 10翻译起始位点代表一个功能性的TATA-少启动子。在这个区域内，我们确定并检查了几个单核苷酸多态性，但没有发现显着差异，在他们的外观之间的AD和非痴呆对照组。通过缺失分析，定点突变，转录因子过表达和电泳迁移率变动分析，我们确定了核苷酸-508到-300为核心启动子，并发现Sp1，USF，和视黄酸响应元件来调节其活性。最后，我们确定了维生素A酸（RA）作为人类ADAM 10启动子活性的诱导剂。这一发现表明，RA受体的药理学靶向可能增加α-分泌酶ADAM 10的表达，对AD病理学具有有益作用。
The ADAM10 gene encodes a membrane-bound disintegrin-metalloproteinase, which, after overexpression in an Alzheimer disease ( AD) mouse model, prevents amyloid pathology and improves long-term potentiation and memory. Because enhancing ADAM10 expression appears to be a reasonable approach for treatment of AD, we functionally analyzed the ADAM10 gene. Both human and mouse ADAM10 genes comprise similar to 160 kbp, are composed of 16 exons, and are evolutionarily highly conserved within 500 bp upstream of either translation initiation site. By using luciferase reporter assays, we demonstrate that nucleotides - 2179 to - 1 upstream of the human ADAM10 translation initiation site represent a functional TATA-less promoter. Within this region we identified and examined several single nucleotide polymorphisms, but did not detect significant differences in their appearance between AD and nondemented control subjects. By deletion analysis, site-directed mutagenesis, transcription factor overexpression and electrophoretic mobility shift assays, we identified nucleotides - 508 to - 300 as the core promoter and found Sp1, USF, and retinoic acid-responsive elements to modulate its activity. Finally, we identified vitamin A acid (RA) as an inducer of human ADAM10 promoter activity. This finding suggests that pharmacologic targeting of RA receptors may increase the expression of the alpha-secretase ADAM10 with beneficial effects on AD pathology.