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Are All DNA Binding and Transcription Regulation by an Activator Physiologically Relevant?

机译：激活剂在生理上是否与所有DNA结合和转录调控有关？

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Understanding how a regulatory protein occupies its sites in vivo is central to understanding gene regulation. Using the yeast Gal4 protein as a model for such studies, we have found 239 potential Gal4 binding sites in the yeast genome, 186 of which are in open reading frames (ORFs). This raises the questions of whether these sites are occupied by Gal4 and, if so, to what effect. We have analyzed theSaccharomyces cerevisiae ACC1 gene (encoding acetyl-coenzyme A carboxylase), which has three Gal4 binding sites in its ORF. The plasmid titration assay has demonstrated that Gal4 occupies these sites in the context of an active ACC1gene. We also find that the expression of the ACC1 is reduced fourfold in galactose medium and that this reduction is dependent on the Gal4 binding sites, suggesting that Gal4 bound to the ORF sites affects transcription of ACC1. Interestingly, removal of the Gal4 binding sites has no obvious effect on the growth in galactose under laboratory conditions. In addition, though the sequence of the ACC1 gene is highly conserved among yeast species, these Gal4 binding sites are not present in theKluyveromyces lactis ACC1 gene. We suggest that the occurrence of these sites may not be related to galactose regulation and a manifestation of the “noise” in the occurrence of Gal4 binding sites.
机译：了解调节蛋白如何在体内占据其位点是了解基因调节的关键。使用酵母Gal4蛋白作为此类研究的模型，我们在酵母基因组中发现了239个潜在的Gal4结合位点，其中186个处于开放阅读框（ORF）中。这就提出了一个问题，即这些位点是否被Gal4所占据，如果是的话，其作用是什么。我们分析了啤酒酵母ACC1 基因（编码乙酰辅酶A羧化酶），该基因在其ORF中具有三个Gal4结合位点。质粒滴定分析表明，在活跃的 ACC1 基因中，Gal4占据了这些位点。我们还发现， ACC1 的表达在半乳糖培养基中降低了四倍，并且这种降低取决于Gal4结合位点，这表明与ORF位点结合的Gal4影响了 ACC1 < / em>。有趣的是，在实验室条件下去除Gal4结合位点对半乳糖的生长没有明显影响。此外，尽管 ACC1 基因的序列在酵母菌种中高度保守，但这些Gal4结合位点在乳酸克鲁维酵母ACC1 基因中并不存在。我们建议这些位点的出现可能与半乳糖调节和Gal4结合位点的出现中的“噪音”的出现无关。

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