The histone deacetylase inhibitor trichostatin A mediates upregulation of 5-lipoxygenase promoter activity by recruitment of Sp1 to distinct GC-boxes
The histone deacetylase inhibitor trichostatin A mediates upregulation of 5-lipoxygenase promoter activity by recruitment of Sp1 to distinct GC-boxes
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DOI：
10.1016/j.bbalip.2007.08.003
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发表时间：
2007-10-01
影响因子：
4.8
通讯作者：
Steinhilber, Dieter
中科院分区：
文献类型：
--
作者：
Schnur, Nicole;Seuter, Sabine;Steinhilber, Dieter
历史上的去乙酰化酶抑制剂阿替他汀A（TsA）在报告基因测定中有效诱导5-脂氧合酶（5-LO）启动子活性以及5-LO mRNA表达。我们在5-LO阴性的HeLa细胞和5-LO表达的Mono Mac 6（MM 6）细胞中鉴定了5-LO基因启动子中介导TsA效应的两个近端Sp1/Sp3结合位点，相比之下，串联GC盒对TsA效应并不重要。TsA既不改变Sp1/Sp3的蛋白质表达水平，也不改变历史上的脱乙酰酶HDAC 1/2的蛋白质表达水平，也不明显改变这些因子形成的蛋白质复合物。此外，用TsA处理细胞没有改变细胞提取物中Sp1/Sp3的结合亲和力，如通过使用含有近端GC盒的探针的达帕分析所测试的。然而，在活细胞中，TsA诱导Sp1、Sp3和RNA聚合酶11募集到5-LO启动子，而不改变历史蛋白H4的乙酰化状态。共转染研究表明，Sp1和Sp3都可以介导TsA效应。这是第一份证明Sp3参与5-LO启动子活性调节的报告。总之，我们表明，TsA增加5-LO启动子活性的增强招聘的Sp1和Sp3的5-LO启动子。(c)2007 Elsevier B. V.保留所有权利。
The historic deacetylase inhibitor trichostatin A (TsA) potently induces 5-lipoxygenase (5-LO) promoter activity in reporter gene assays as well as 5-LO mRNA expression. We identified two proximal Sp1/Sp3 binding sites in the 5-LO gene promoter mediating the TsA effect in both 5-LO-negative HeLa cells and in 5-LO expressing Mono Mac 6 (MM6) cells, the tandem GC-boxes, by contrast, were not important for the TsA effect. TsA neither altered the protein expression levels of Sp1/Sp3 nor of the historic deacetylases HDAC1/2, nor did it apparently change the protein complex formation by these factors. Also, treatment of cells with TsA did not change the binding affinity of Sp1/Sp3 in cell extracts, as tested by DAPA analysis using probes containing the proximal GC boxes. However, in the living cell TsA induced Sp1, Sp3 and RNA polymerase 11 recruitment to the 5-LO promoter without changing the acetylation status of historic protein H4. Cotransfection studies suggest that both Sp1 and Sp3 can mediate the TsA effect. This is the first report demonstrating that Sp3 is involved in the regulation of 5-LO promoter activity. In summary, we show that TsA increases 5-LO promoter activity by the enhanced recruitment of Sp1 and Sp3 to the 5-LO promoter. (c) 2007 Elsevier B.V. All rights reserved.