Role of Ets factors in the activity and endothelial cell specificity of the mouse Tie gene promoter
Role of Ets factors in the activity and endothelial cell specificity of the mouse Tie gene promoter
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DOI：
10.1096/fasebj.13.2.377
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发表时间：
1999-02-01
期刊：
影响因子：
4.8
通讯作者：
Alitalo, K
中科院分区：
文献类型：
--
作者：
Iljin, K;Dube, A;Alitalo, K
Tie基因编码正常血管发育所必需的内皮细胞受体酪氨酸激酶。Tie基因启动子靶向转基因小鼠中特异性针对内皮细胞的异源基因表达。在这里，我们的特点是在培养细胞和转基因小鼠的内皮细胞特异性活性的启动子序列的关键。启动子的逐步缺失和定点突变表明，内皮细胞特异性的关键元件是一个八聚体转录因子结合位点和位于主要转录起始位点上游300 bp内的两个簇中的几个Ets结合位点。（一种与ets因子ELF-1相关的新型转录因子），其在内皮细胞中表达，ETS 2显示Tie启动子的最强反式激活; ETS 1提供较低水平的刺激，而其他Ets因子提供很少或没有反式激活。Tie启动子引导大量人生长激素的产生进入转基因小鼠的循环，分泌量与转基因拷贝数相关，对转基因整合位点的作用相对不敏感，这些性质表明Tie启动子活性受内皮细胞Ets因子控制，并且其具有用于内皮细胞特异性基因表达的载体的潜力。
The Tie gene encodes an endothelial cell receptor tyrosine kinase necessary for normal vascular development, The Tie gene promoter targets expression of heterologous genes specifically to endothelial cells in transgenic mice. Here we have characterized the promoter sequences critical for endothelial cell-specific activity in cultured cells and transgenic mice. Progressive deletions and site-directed mutations of the promoter showed that the critical endothelial cell-specific elements are an octamer transcription factor binding site and several Ets binding sites located in two clusters within 300 bp upstream of the major transcription initiation site, Among members of the Ets transcription factor family tested, NERF-2 (a novel transcription factor related to the ets factor ELF-1), which is expressed in endothelial cells, and ETS2 showed the strongest transactivation of the Tie promoter; ETS1 gave lower levels of stimulation and the other Ets factors gave little or no transactivation, Furthermore, the Tie promoter directed the production of high amounts of human growth hormone into the circulation of transgenic mice, The secreted amounts correlated with transgene copy number, being relatively insensitive to the effects of the transgene integration site, These properties suggest that Tie promoter activity is controlled by endothelial cell Ets factors and that it has potential for use in vectors for endothelial cell-specific gene expression.