McKay L M, Carpenter B, Roberts S G
Division of Gene Expression, Department of Biochemistry, Wellcome Trust Building, University of Dundee, Dundee DD1 5EH, UK.
Nucleic Acids Res. 1999 Aug 1;27(15):3064-70. doi: 10.1093/nar/27.15.3064.
Even-skipped (Eve) is a transcriptional repressor involved in segment formation in Drosophila melano-gaster. In order to gain further insights into the mechanism of action of Eve we tested whether it would function as a transcriptional repressor in mammalian cells. We found that Eve was indeed a potent repressor in two different mammalian cell types and at several promoters. In vitro transcription assays confirmed that Eve directly represses transcription initiation when specifically targeted to a promoter. We also found that, unlike the case with transcriptional activators, Eve does not repress transcription synergistically. Analysis of the effect of Eve on preinitiation complex assembly in a crude HeLa cell nuclear extract demonstrated that the Eve repression domain functions by preventing the assembly of TFIID with the promoter. Our data support the hypothesis that Eve contains an active repression domain that functions specifically to prevent preinitiation complex formation.
偶数跳格基因（Eve）是一种转录抑制因子，参与黑腹果蝇的体节形成。为了进一步深入了解Eve的作用机制，我们测试了它在哺乳动物细胞中是否会作为转录抑制因子发挥作用。我们发现Eve在两种不同的哺乳动物细胞类型以及多个启动子处确实是一种有效的抑制因子。体外转录分析证实，当Eve特异性靶向启动子时，它会直接抑制转录起始。我们还发现，与转录激活因子的情况不同，Eve不会协同抑制转录。对Eve在粗制的HeLa细胞核提取物中对起始前复合物组装的影响进行分析表明，Eve抑制结构域通过阻止TFIID与启动子组装来发挥作用。我们的数据支持这样一种假设，即Eve含有一个活性抑制结构域，其功能是专门阻止起始前复合物的形成。
