DIFFERENT DOMAINS REGULATE THE HUMAN ERYTHROPOIETIN RECEPTOR GENE-TRANSCRIPTION
DIFFERENT DOMAINS REGULATE THE HUMAN ERYTHROPOIETIN RECEPTOR GENE-TRANSCRIPTION
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DOI：
10.1093/nar/22.3.338
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发表时间：
1994-02-11
影响因子：
14.9
通讯作者：
CHRETIEN, S
中科院分区：
文献类型：
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作者：
MAOUCHE, L;CARTRON, JP;CHRETIEN, S
为了分析人促红细胞生成素受体（hEpo-R）基因的组织特异性转录所需的5 '侧翼序列，探索了相对于转录起始位点（+1）跨越核苷酸-1050至+135的DNA区域。我们的研究表明，在-45和-20位含有加塔和SP1结合位点的最小启动子（-76/+33）不足以赋予报告基因红系特异性表达。用含有一簇Alu重复元件的（-1050/+33构建体）或在转录起始位点下添加135 bp（-76/+135构建体）观察到启动子的红系特异性，所述构建体对启动子活性施加负控制，在非红系组织中具有主要作用。后一个区域可以细分为两个不同的结构域：+1/+78区域发挥积极作用，+79/+135区域对CAT测定测量的Epo-R启动子活性具有负面影响。第一个区域含有三个CANNTG基序，而第二个区域在+85位含有SP1/CACCC基序。这些发现揭示了一个复杂的调控hEpo-R基因，并提供了一个工作模型，用于解释如何最小的启动子，含有加塔/SP1，可以积极和消极的调节在红细胞分化。
To analyse the 5'-flanking sequences required for the tissue specific transcription of the human erythropoietin receptor (hEpo-R) gene, a DNA region spanning nucleotides -1050 to +135 relative to the transcription initiation site (+1) was explored. Our studies indicate that a minimum promoter (-76/+33) containing GATA and SP1 binding sites at positions -45 and -20 is not sufficient to confer erythroid specific expression to a reporter gene. Erythroid specificity of the promoter was observed either with the (-1050/+33 construct) which contains a cluster of Alu repetitive elements or with the addition of the 135 bp down to the transcription initiation site (-76/+135 construct) which exert a negative control on the promoter activity with a major effect in non erythroid tissues. The latter region can be subdivided on two distinct domains: the +1/+78 region that exerts a positive effect and the +79/+135 region that has a negative effect on the Epo-R promoter activity measured by CAT assays. The first region contains three CANNTG motifs, whereas the second contains an SP1/CACCC motif at position +85. These findings reveal a complex regulation of the hEpo-R gene and provide a working model useful to explain how the minimal promoter, containing GATA/SP1, can be positively and negatively regulated during erythroid differentiation.