Transcription of human ABO histo-blood group genes is dependent upon binding of transcription factor CBF/NF-Y to minisatellite sequence
Transcription of human ABO histo-blood group genes is dependent upon binding of transcription factor CBF/NF-Y to minisatellite sequence
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DOI：
10.1074/jbc.272.41.25890
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发表时间：
1997-10-10
影响因子：
4.8
通讯作者：
Yamamoto, F
中科院分区：
文献类型：
--
作者：
Kominato, Y;Tsuchiya, T;Yamamoto, F
我们研究了人类组织血型ABO基因的转录调控机制，并鉴定了控制这些基因表达的DNA顺式元件和反式激活蛋白，这些基因在输血和器官移植中具有重要意义。我们将ABO基因的5 '-上游序列引入到无启动子的报告载体中，并使用胃癌细胞系KATO III细胞的瞬时转染试验来表征缺失构建体的启动子活性。在胃癌细胞中，转录起始位点（帽位点）上游的序列和增强子元件（位于转录起始位点上游-3899和-3618碱基对（bp）之间）并含有4个串联拷贝的43-bp重复单位）被证明负责ABO基因的转录活性。DNA结合研究表明，转录因子CBF/NF-Y与小卫星中的43-bp重复单元结合。这些CBF/NF-Y结合位点在增强子活性中的功能重要性通过使用具有突变结合位点的报告质粒的转染实验来证实。因此，人ABO基因的转录调节依赖于CBF/NF-Y与小卫星的结合。
We have studied the transcriptional regulatory mechanism of the human histo-blood group ABO genes, and identified DNA cis-elements and trans-activating protein that control the expression of these genes which are important in blood transfusion and organ transplantation. We introduced the 5'-upstream sequence of ABO genes into the promoterless reporter vector and characterized the promoter activity of deletion constructs using transient transfection assays with gastric cancer cell line KATO III cells. The sequence just upstream of the transcription start site (cap site), and an enhancer element, which is located further upstream (between -3899 and -3618 base pairs (bp) from the transcription initiation site) and contains 4 tandem copies of a 43-bp repeat unit, were shown in gastric cancer cells to be responsible for the transcriptional activity of the ABO genes. DNA binding studies have demonstrated that a transcription factor, CBF/NF-Y, bound to the 43-bp repeat unit in the minisatellite. Functional importance of these CBF/NF-Y-binding sites in enhancer activity was confirmed by transfection experiments using reporter plasmids with mutated binding sites. Thus, transcriptional regulation of the human ABO genes is dependent upon binding of CBF/NF-Y to the minisatellite.