A GABP-binding element in the Robo4 promoter is necessary for endothelial expression in vivo
A GABP-binding element in the Robo4 promoter is necessary for endothelial expression in vivo
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DOI：
10.1182/blood-2008-01-135079
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发表时间：
2008-09-15
期刊：
影响因子：
20.3
通讯作者：
Aird, William C.
中科院分区：
文献类型：
--
作者：
Okada, Yoshiaki;Jin, Enjing;Aird, William C.
我们最近证实，人ROBO4基因的3-kb 5‘-侧翼区在体外和体内都能引导内皮细胞特异性表达。此外，-119处的GA结合蛋白(GABP)结合基序是介导体外启动子活性所必需的。本研究的目的是确认-119 GABP结合位点在体内的功能相关性。为此，用一个带有GABP突变位点的Robo4-LacZ转基因盒来定位小鼠的HPRT基因。在其他研究中，GABP突变被引入到敲入LacZ的内源性小鼠Robo4基因座中。与各自的对照相比，突变启动子在类胚体、胚胎和成年动物中的活性显著降低。综上所述，这些数据为GABP结合基序在调节完整内皮细胞Robo4表达中的作用提供了强有力的支持。
We recently demonstrated that the 3-kb 5'-flanking region of the human ROBO4 gene directs endothelial cell-specific expression in vitro and in vivo. Moreover, a GA-binding protein (GABP)-binding motif at -119 was necessary for mediating promoter activity in vitro. The goal of the present study was to confirm the functional relevance of the -119 GABP-binding site in vivo. To that end, the Hprt locus of mice was targeted with a Robo4-LacZ transgenic cassette in which the GABP site was mutated. In other studies, the GABP mutation was introduced into the endogenous mouse Robo4 locus in which LacZ was knocked-in. Compared with their respective controls, the mutant promoters displayed a significant reduction in activity in embryoid bodies, embryos, and adult animals. Together, these data provide strong support for the role of the GABP-binding motif in mediating Robo4 expression in the intact endothelium.