STAT1 transcriptionally regulates the expression of S1PR1 by binding its promoter region
STAT1 transcriptionally regulates the expression of S1PR1 by binding its promoter region
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STAT1 通过结合 S1PR1 的启动子区域来转录调节 S1PR1 的表达
DOI：
10.1016/j.gene.2020.144417
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发表时间：
2020
期刊：
影响因子：
3.5
通讯作者：
Liu Qiji
中科院分区：
文献类型：
--
作者：
Xin Qian;Cheng Guanghui;Kong Feng;Ji Qinghong;Li Haizhen;Jiang Wen;Wang Jue;Luan Yun;Sun Chao;Chen Xiaojing;Wang Xuexiang;Qi Tonggang;Xu Dawei;Song Lei;Liu Qiji
鞘氨醇1-磷酸受体1（S1 PR 1）在介导免疫细胞的运输和迁移中起关键作用。以往的研究也证实S1 PR 1是哮喘等自身免疫性疾病的重要易感基因。然而，对S1 PR 1表达的调控机制知之甚少。因此，本研究系统地研究了S1 PR 1的转录调控。使用一系列pGL 3-Basic报告载体仔细筛选S1 PR 1基因的启动子活性，所述报告载体包含全长（范围从转录起始位点到上游-1 kb区域）或几个截短的S1 PR 1启动子片段。我们确定了S1 PR 1启动子的一个区域（从-29到-12 bp）作为最小启动子区域。生物信息学预测结果表明，几个转录因子被招募到这些网站。EMSA和ChIP检测证实转录因子STAT 1可与该区域结合。我们还发现，当STAT 1被敲低时，S1 PR 1水平显著降低。与STAT 1耗竭引起的S1 PR 1减少一致，STAT 1过表达导致S1 PR 1上调。另外，IFN-γ激活STAT 1后，S1 PR 1的mRNA和蛋白水平均升高，而氟达拉滨抑制STAT 1后，S1 PR 1的mRNA和蛋白水平均降低。此外，STAT 1和S1 PR 1的表达水平在41名健康人的外周血白细胞中呈正相关。我们的研究表明，转录因子STAT 1可以结合到S1 PR 1启动子上游-29 bp到-12 bp的区域，并刺激S1 PR 1的表达。
Sphingosine 1-phosphate receptor 1 (S1PR1) plays a pivotal role in mediating trafficking and migration of immune cells. Previous reports also identifyS1PR1as an important susceptibility gene of asthma and other autoimmune disorders. However, little has been known about the regulatory mechanism of S1PR1 expression. Thus we systematically investigated the transcriptional regulation of S1PR1 in this study. Promoter activity ofS1PR1gene was carefully screened using series of pGL3-Basic reporter vectors, containing full length (range from transcription start site to upstream −1 kb region) or several truncated fragments ofS1PR1promoter. We identified an area (from −29 to −12 bp) of theS1PR1promoter as the minimal promoter region. Bioinformatics prediction results showed that several transcription factors were recruited to these sites. EMSA and ChIP assays demonstrated the transcriptional factor STAT1 could bind to the region. We also found that the level of S1PR1 level was significantly reduced when STAT1 was knocked-down. Consistent with the reduction of S1PR1 caused by depletion of STAT1, overexpression of STAT1 resulted in up-regulation of S1PR1. In addition, both mRNA and protein levels of S1PR1 were increased when STAT1 was activated by IFN-γ, and decreased when STAT1 was inhibited by fludarabine. Besides, the levels of STAT1 and S1PR1 expression were positively correlated in peripheral blood leukocytes derived from 41 healthy individuals. Our study showed that transcription factor STAT1 could bind to upstream region of −29 bp to −12 bp of theS1PR1promoter and stimulate the expression of S1PR1.