Histone deacetylase inhibitors stimulate mitochondrial HMG-CoA synthase gene expression via a promoter proximal Sp1 site
Histone deacetylase inhibitors stimulate mitochondrial HMG-CoA synthase gene expression via a promoter proximal Sp1 site
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DOI：
10.1093/nar/gkg262
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发表时间：
2003-03-15
影响因子：
14.9
通讯作者：
Marrero, PF
中科院分区：
文献类型：
--
作者：
Camarero, N;Nadal, A;Marrero, PF
线粒体HMG-CoA合酶在结肠中的表达与该组织中存在的丁酸水平相关。我们在这里报告丁酸对线粒体HMG-CoA合酶基因表达的影响是在体内转录水平上发挥作用的，并且一种特异性组蛋白去乙酰化酶抑制剂-丁酸他汀A（TSA）也诱导了来自结肠癌的人细胞系中该基因的转录活性和mRNA表达。使用染色质免疫沉淀试验，我们表明，组蛋白脱乙酰酶1（HDAC 1）与内源性线粒体HMG-CoA合成酶启动子和TSA诱导与H4组蛋白的超乙酰化与5'侧翼区的基因。HDAC 1活性的过表达一致地导致线粒体HMG-CoA合酶启动子低乙酰化并降低其转录活性。丁酸盐和TSA的作用映射到基因近端启动子中存在的单个Sp1位点，该位点能够结合Sp1和Sp3蛋白。有趣的是，Sp1和Sp3蛋白与Sp1位点的结合亲和力与启动子的TSA响应性相关。使用单杂交系统（GAL 4-Sp1和GAL 4-Sp3），我们表明，这两种蛋白质可以介导TSA在CaCo-2细胞采用不同的机制。
The expression of mitochondrial HMG-CoA synthase in the colon has been correlated with the levels of butyrate present in this tissue. We report here that the effect of butyrate on mitochondrial HMG-CoA synthase gene expression is exerted in vivo at the transcriptional level, and that trichostatin A (TSA), a specific histone deacetylase inhibitor, also induces transcriptional activity and mRNA expression of the gene in human cell lines derived from colon carcinoma. Using chromatin immunoprecipitation assays, we show that histone deacetylase 1 (HDAC1) is associated with the endogenous mitochondrial HMG-CoA synthase promoter and that TSA induction correlates with hyperacetylation of H4 histone associated with the 5' flanking region of the gene. Overexpression of HDAC1 activity leads consistently to mitochondrial HMG-CoA synthase promoter hypoacetylation and reduces its transcriptional activity. The effect of butyrate and TSA maps to a single Sp1 site present in the proximal promoter of the gene, which is able to bind Sp1 and Sp3 proteins. Interestingly, the binding affinity of Sp1 and Sp3 proteins to the Sp1 site correlates with the TSA responsiveness of the promoter. Using a one-hybrid system (GAL4-Sp1 and GAL4-Sp3), we show that both proteins can mediate responsiveness to TSA in CaCo-2 cells employing distinct mechanisms.