Functional domains of the yeast STE12 protein, a pheromone-responsive transcriptional activator.
Functional domains of the yeast STE12 protein, a pheromone-responsive transcriptional activator.
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酵母 STE12 蛋白的功能域，一种信息素响应转录激活剂。
DOI：
10.1128/mcb.13.6.3765-3772.1993
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发表时间：
1993
影响因子：
5.3
通讯作者：
Fields,S
中科院分区：
文献类型：
--
作者：
Kirkman-Correia,C;Stroke,IL;Fields,S
酵母Saccharomyces goulae的信息素反应途径是细胞类型特异性基因转录的基础水平以及信息素处理后观察到的诱导水平所必需的。STE 12蛋白与指定为信息素反应元件的DNA序列结合，并且是信息素诱导的信号的靶标。我们构建了STE 12的6-核苷酸接头插入突变体、内部缺失突变体和羧基末端截短突变体，并测定了它们恢复aste 12 Δ菌株交配和转录活性的能力。这些突变体蛋白质中的两个保留了介导基础转录的能力，但在信息素处理后显示很少或没有诱导转录。产生这些蛋白质的细胞不能交配，正式证明了通过增加基因表达来响应信息素的能力对于交配过程是必不可少的。由于STE 12的不同结构域似乎是基础与诱导转录所需的，我们建议，信息素诱导的信号很可能靶向的蛋白质的残基不同于基础信号靶向的那些残基，因为响应途径的组成性活性。我们对突变型STE 12蛋白的分析还表明，只有DNA结合结构域对接头插入引起的微小变化敏感。此外，我们表明，虽然STE 12与转录因子MCM 1形成复合物所必需的羧基末端序列对于交配不是必需的，但这些序列是最佳转录活性所必需的。
The pheromone response pathway of the yeastSaccharomyces cerevisiaeis necessary for the basal level of transcription of cell-type-specific genes, as well as the induced level observed after pheromone treatment. The STE12 protein binds to the DNA sequence designated the pheromone response element and is a target of the pheromone-induced signal. We generated 6-nucleotide linker insertion mutants, internal-deletion mutants, and carboxy-terminal truncation mutants of STE12 and assayed them for their ability to restore mating and transcriptional activity to aste12Δ strain. Two of these mutant proteins retain the capacity to mediate basal transcription but show little or no induced transcription upon pheromone treatment. Cells producing these proteins cannot mate, formally demonstrating that the ability to respond to pheromone by increasing gene expression is essential for the mating process. Since distinct domains of STE12 appear to be required for basal versus induced transcription, we suggest that the pheromone-induced signal is likely to target residues of the protein different from those targeted by the basal signal because of the constitutive activity of the response pathway. Our analysis of mutant STE12 proteins also indicates that only the DNA-binding domain is sensitive to the small changes caused by the linker insertions. In addition, we show that, while the carboxy-terminal sequences necessary for STE12 to form a complex with the transcription factor MCM1 are not essential for mating, these sequences are required for optimal transcriptional activity.