Dolan J W, Kirkman C, Fields S
Department of Microbiology, State University of New York, Stony Brook 11794.
Proc Natl Acad Sci U S A. 1989 Aug;86(15):5703-7. doi: 10.1073/pnas.86.15.5703.
The STE12 gene product of Saccharomyces cerevisiae is required for the transcription of two sets of cell-type-specific genes: the a-specific genes (active only in a cells) and the alpha-specific genes (active only in alpha cells). We show that radiolabeled STE12 protein, prepared by in vitro transcription and translation, is capable of forming complexes with unlabeled DNA fragments from two a-specific genes. Wild-type yeast, but not a ste12 mutant, produce a factor that forms complexes with labeled DNA from these same two genes. We use assays with yeast extracts to localize the binding site for the STE12-dependent activity. This site corresponds to the sequence identified as the pheromone induction element, which is responsible for increased transcription of genes when cells are exposed to alpha-factor or a-factor. Thus the STE12 protein may be an ultimate effector in the signal transduction pathway triggered by pheromone.
酿酒酵母的STE12基因产物是两组细胞类型特异性基因转录所必需的：a特异性基因（仅在a细胞中活跃）和α特异性基因（仅在α细胞中活跃）。我们表明，通过体外转录和翻译制备的放射性标记的STE12蛋白能够与来自两个a特异性基因的未标记DNA片段形成复合物。野生型酵母而非ste12突变体产生一种因子，该因子能与来自这两个相同基因的标记DNA形成复合物。我们利用酵母提取物检测来定位STE12依赖性活性的结合位点。该位点对应于被鉴定为信息素诱导元件的序列，当细胞暴露于α因子或a因子时，该元件负责基因转录的增加。因此，STE12蛋白可能是信息素触发的信号转导途径中的最终效应物。
