We analysed the genome-wide regulatory properties of an artificial transcription activator in which the DNA-binding domain of the yeast transcription factor, Pdr1, was fused to the activation domain of Gal4 (Pdr1*GAD). This Pdr1*GAD chimera was put under the control of the inducible GAL1 promoter. DNA microarray analyses showed that all the target genes upregulated by the well-studied native gain-of-function Pdr1-3 mutant were similarly activated by the chimerical factor Pdr1*GAD upon galactose induction. Additionally, this kinetic approach led us not only to confirm previously published targets, but also to define a hierarchy among members of the Pdr1 regulon. Our observations prove, for the first time at the complete genome level, that the DNA-binding domain of Pdr1 is sufficient to guide its specificity. We propose that this approach could be useful for the study of new transcription factors identified in silico from sequenced organisms. Complete data are available at www.biologie.ens.fr/yeast-publi.html.

译文

我们分析了人工转录激活因子的全基因组调控特性,其中酵母转录因子Pdr1的DNA结合结构域与Gal4 (Pdr1 * GAD) 的激活结构域融合。将该Pdr1 * GAD嵌合体置于诱导型GAL1启动子的控制下。DNA微阵列分析表明,经过充分研究的天然功能获得Pdr1-3突变体上调的所有靶基因在半乳糖诱导后被嵌合因子Pdr1 * GAD类似地激活。此外,这种动力学方法不仅使我们确认了先前发布的目标,而且还定义了Pdr1正则子成员之间的层次结构。我们的观察首次在完整基因组水平上证明,Pdr1的DNA结合域足以指导其特异性。我们建议这种方法可用于研究从测序生物在计算机中鉴定的新转录因子。完整的数据可在www.biologie.ens.fr/yeast-publi.html获得。

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