BEST1 expression in the retinal pigment epithelium is modulated by OTX family members.
BEST1 expression in the retinal pigment epithelium is modulated by OTX family members.
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DOI：
10.1093/hmg/ddn323
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发表时间：
2009-01-01
影响因子：
3.5
通讯作者：
Zack DJ
中科院分区：
文献类型：
--
作者：
Esumi N;Kachi S;Hackler L Jr;Masuda T;Yang Z;Campochiaro PA;Zack DJ
视网膜色素上皮（retinal pigment epithelium，RPE）中的许多基因与视网膜变性相关。其中之一，BEST 1，编码bestrophin-1，一种突变时导致Best黄斑营养不良的蛋白质。作为RPE基因调控的模型，我们一直在研究控制BEST 1表达的机制，最近证明MITF-TFE家族的成员调节BEST 1转录。人BEST 1上游区从-154到+38 bp足以指导RPE中的表达，并且正调控元件存在于-154和-104 bp之间。在这里，我们表明，-154至-104 bp的区域是必要的RPE在转基因小鼠的表达，并包含一个预测的OTX结合位点（网站1）。由于另一个非典型OTX位点（位点2）位于附近，我们使用BEST 1启动子/荧光素酶构建体通过体内电穿孔测试了这些位点的功能，发现两个位点的突变降低了启动子活性。三种OTX家族蛋白，OTX 1，OTX 2和CRX，在体外结合位点1和2，并且都增加BEST 1启动子活性。令人惊讶的是，我们发现人和牛RPE不仅表达OTX 2，而且表达CRX，牛RPE中的CRX基因组区域对DNA酶I高度敏感，与活性转录一致，并且OTX 2和CRX在体内都结合到BEST 1近端启动子。这些结果首次证明了CRX在RPE中的表达，并表明OTX 2和CRX可能在RPE中充当BEST 1启动子的正调节剂。
A number of genes preferentially expressed in the retinal pigment epithelium (RPE) are associated with retinal degenerative disease. One of these, BEST1, encodes bestrophin-1, a protein that when mutated causes Best macular dystrophy. As a model for RPE gene regulation, we have been studying the mechanisms that control BEST1 expression, and recently demonstrated that members of the MITF-TFE family modulate BEST1 transcription. The human BEST1 upstream region from -154 to +38 bp is sufficient to direct expression in the RPE, and positive regulatory elements exist between -154 and -104 bp. Here, we show that the -154 to - 104 bp region is necessary for RPE expression in transgenic mice and contains a predicted OTX-binding site (Site 1). Since another non-canonical OTX site (Site 2) is located nearby, we tested the function of these sites using BEST1 promoter/luciferase constructs by in vivo electroporation and found that mutation of both sites reduces promoter activity. Three OTX family proteins, OTX1, OTX2, and CRX, bound to both Sites 1 and 2 in vitro, and all increased BEST1 promoter activity. Surprisingly we found that human and bovine RPE expressed not only OTX2 but also CRX, the CRX genomic region in bovine RPE was hypersensitive to DNase I, consistent with active transcription, and that both OTX2 and CRX bound to the BEST1 proximal promoter in vivo. These results demonstrate for the first time CRX expression in the RPE, and suggest that OTX2 and CRX may act as positive modulators of the BEST1 promoter in the RPE.
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影响因子：
16.2
作者：
Chen, SM;Wang, QL;Zack, DJ
通讯作者：
Zack, DJ
影响因子：
64.5
作者：
Furukawa, T;Morrow, EM;Cepko, CL
通讯作者：
Cepko, CL
影响因子：
48
作者：
Dorschner, M;Hawrylycz, M;Stamatoyannopoulos, JA
通讯作者：
Stamatoyannopoulos, JA
DOI：
10.1073/pnas.73.9.3278
发表时间：
1976-01-01
影响因子：
11.1
作者：
CAREY, TE;TAKAHASHI, T;OLD, LJ
通讯作者：
OLD, LJ
DOI：
10.1016/s0169-328x(00)00060-7
发表时间：
2000-05-31
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Baas, D;Bumsted, KM;Barnstable, CJ
通讯作者：
Barnstable, CJ