SATURATION MUTAGENESIS OF THE YEAST HIS3 REGULATORY SITE - REQUIREMENTS FOR TRANSCRIPTIONAL INDUCTION AND FOR BINDING BY GCN4 ACTIVATOR PROTEIN
SATURATION MUTAGENESIS OF THE YEAST HIS3 REGULATORY SITE - REQUIREMENTS FOR TRANSCRIPTIONAL INDUCTION AND FOR BINDING BY GCN4 ACTIVATOR PROTEIN
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DOI：
10.1126/science.3532321
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发表时间：
1986-10-24
期刊：
影响因子：
56.9
通讯作者：
STRUHL, K
中科院分区：
文献类型：
--
作者：
HILL, DE;HOPE, IA;STRUHL, K
酵母his3和其他氨基酸生物合成基因的表达在氨基酸饥饿条件下被诱导。这种反应的协调是由一种称为 GCN4 的正调控蛋白介导的，该蛋白特异性结合所有共调控基因上游的调控位点并刺激其转录。 his3 调控位点的核苷酸序列要求是通过分析通过克隆寡核苷酸的新方法获得的大量点突变来确定的。九个碱基对序列 ATGACTCTT 内的几乎所有单碱基对突变均显着降低体内 his3 诱导和体外 GCN4 结合，而该区域之外的变化影响最小。一种突变会产生与 15 个共调控基因的共有序列最相似的序列，从而提高了 GCN4 蛋白的诱导水平和亲和力。最佳序列 ATGACTCAT 的回文性质表明 GCN4 蛋白作为二聚体与可能重叠的相邻半位点结合。
Expression of the yeast his3 and other amino acid biosynthetic genes is induced during conditions of amino acid starvation. The coordination of this response is mediated by a positive regulatory protein called GCN4, which binds specifically to regulatory sites upstream of all coregulated genes and stimulates their transcription. The nucleotide sequence requirements of the his3 regulatory site were determined by analysis of numerous point mutations obtained by a novel method of cloning oligonucleotides. Almost all single base pair mutations within the nine base pair sequence ATGACTCTT significantly reduce his3 induction in vivo and GCN4 binding in vitro, whereas changes outside this region have minimal effects. One mutation, which generates a sequence that most closely resembles the consensus for 15 coregulated genes, increases both the level of induction and the affinity for GCN4 protein. The palindromic nature of the optimal sequence, ATGACTCAT, suggests that GCN4 protein binds as a dimer to adjacent half-sites that possibly overlap.