Pheromone-dependent phosphorylation of the yeast STE12 protein correlates with transcriptional activation.
Pheromone-dependent phosphorylation of the yeast STE12 protein correlates with transcriptional activation.
复制标题
酵母 STE12 蛋白的信息素依赖性磷酸化与转录激活相关。
DOI：
10.1101/gad.5.5.741
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发表时间：
1991
影响因子：
10.5
通讯作者：
Fields,S
中科院分区：
文献类型：
--
作者：
Song,D;Dolan,JW;Yuan,YL;Fields,S
酵母的单倍体 a 和 α 细胞分别通过增加许多基因的转录来响应信息素 α 和 a 因子，而这些基因的产物对于交配至关重要。 STE12 蛋白在此过程中通过与 DNA 序列结合来发挥作用，从而介导信息素响应基因转录的增加。我们在此表明，含有与 GAL4 DNA 结合结构域融合的 STE12 的杂合蛋白可以激活含有 GAL4 结合位点的报告基因的转录，但只有在用信息素处理细胞后才能激活。因此，当 STE12 单独与 DNA 结合时，就足以介导信息素诱导的转录。通过构建不同 STE12 区域与 GAL4 结构域的杂交体，我们将这种激活所需的 STE12 结构域映射到蛋白质的中央三分之一。经过 α 因子处理后，GAL4 与完整 STE12 序列的杂合体迅速磷酸化，其动力学与观察到的信息素响应基因的转录诱导一致。这种磷酸化所必需的 STE12 结构域与转录激活相关。我们提出，信息素响应基因的诱导是通过 STE12 的磷酸化介导的，从而改变其激活功能，但不改变其 DNA 结合能力。
Haploid a and alpha cells of yeast respond to the pheromones alpha- and a-factor, respectively, by increasing the transcription of many genes whose products are essential for mating. The STE12 protein acts in this process by binding to the DNA sequence that mediates the increased transcription of pheromone-responsive genes. We show here that a hybrid protein containing STE12 fused to the DNA-binding domain of GAL4 can activate transcription of a reporter gene containing GAL4-binding sites but only after treatment of cells with pheromone. Thus, STE12 alone, when bound to DNA, is sufficient to mediate pheromone-induced transcription. By constructing hybrids of different STE12 regions with the GAL4 domain, we map the domain of STE12 necessary for this activation to the central third of the protein. Upon alpha-factor treatment, the hybrid of GAL4 with the complete STE12 sequence is rapidly phosphorylated, with kinetics consistent with the observed transcriptional induction of pheromone-responsive genes. The domain of STE12 necessary for this phosphorylation correlates with that involved in transcriptional activation. We propose that induction of pheromone-responsive genes is mediated by phosphorylation of STE12 to alter its activation function but not its DNA-binding ability.