Transcriptional regulation of the MDR1 gene by histone acetyltransferase and deacetylase is mediated by NF-Y
Transcriptional regulation of the MDR1 gene by histone acetyltransferase and deacetylase is mediated by NF-Y
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DOI：
10.1128/mcb.18.7.4377
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发表时间：
1998-07-01
影响因子：
5.3
通讯作者：
Scotto, KW
中科院分区：
文献类型：
--
作者：
Jin, SK;Scotto, KW
最近的研究表明，组蛋白修饰酶组蛋白乙酰转移酶（HAT）和组蛋白去乙酰化酶（HDAC）分别参与转录激活和抑制。然而，很少有人知道的内源性基因，这些酶的调节或如何实现特异性。在本报告中，我们证明，HAT和HDAC活动调节的P-糖蛋白编码基因，MDR 1的转录。将人结肠癌SW 620细胞在100 ng/ml的HDAC特异性抑制剂阿司他丁A（TSA）中孵育，可使MDR 1 mRNA的稳态水平增加20倍。此外，TSA处理用野生型MDR 1启动子/荧光素酶构建体转染的细胞导致启动子活性的10至15倍诱导。缺失和点突变分析确定，一个倒置的CCAAT盒是必不可少的fdr这种激活。与此观察结果一致，p300/CREB结合蛋白相关因子（P/CAF），一种具有内在HAT活性的转录共激活因子的过表达激活了野生型MDR 1启动子，但不激活CCAAT盒中含有突变的启动子; P/CAF HAT结构域的缺失废除了激活。凝胶位移和supershift分析确定NF-Y的CCAAT盒结合蛋白在这些细胞中，和显性负NF-Y表达载体的共转染减少了TSA的MDR 1启动子的激活。此外，NF-YA和P/CAF在体外显示出相互作用。这是第一个报告的天然启动子，是调制。通过HAT和HDAC活性，其中已经鉴定了介导这种调节的转录因子。
Recent studies have shown that the histone-modifying enzymes histone acetyltransferase (HAT) and histone deacetylase (HDAC) are involved in transcriptional activation and repression, respectively. However, little is known about the endogenous genes that are regulated by these enzymes or how specificity is achieved. In the present report, we demonstrate that HAT and HDAC activities modulate transcription of the P-glycoprotein-encoding gene, MDR1. Incubation of human colon carcinoma SW620 cells in 100-ng/ml trichostatin A (TSA), a specific HDAC inhibitor, increased the steady-state level of MDR1 mRNA 20-fold. Furthermore, TSA treatment of cells transfected with a wild-type MDR1 promoter/luciferase construct resulted in a 10- to 15-fold induction of promoter activity. Deletion and point mutation analysis determined that an inverted CCAAT box was essential fdr this activation. Consistent with this observation, overexpression of p300/CREB binding protein-associated factor (P/CAF), a transcriptional coactivator with intrinsic HAT activity, activated the wild-type MDR1 promoter but not a promoter containing a mutation in the CCAAT box; deletion of the P/CAF HAT domain abolished activation. Gel shift and supershift analyses identified NF-Y as the CCAAT-box binding protein in these cells, and cotransfection of a dominant negative NF-Y expression vector decreased the activation of the MDR1 promoter by TSA. Moreover, NF-YA and P/CAF were shown to interact in vitro. This is the first report of a natural promoter that is modulated. by HAT and HDAC activities in which the transcription factor mediating this regulation has been identified.