Grishin A V, Rothenberg M, Downs M A, Blumer K J
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
Genetics. 1998 Jun;149(2):879-92. doi: 10.1093/genetics/149.2.879.
In the yeast Saccharomyces cerevisiae, mating pheromone response is initiated by activation of a G protein- and mitogen-activated protein (MAP) kinase-dependent signaling pathway and attenuated by several mechanisms that promote adaptation or desensitization. To identify genes whose products negatively regulate pheromone signaling, we screened for mutations that suppress the hyperadaptive phenotype of wild-type cells overexpressing signaling-defective G protein beta subunits. This identified recessive mutations in MOT3, which encodes a nuclear protein with two Cys2-His2 Zn fingers. MOT3 was found to be a dosage-dependent inhibitor of pheromone response and pheromone-induced gene expression and to require an intact signaling pathway to exert its effects. Several results suggested that Mot3 attenuates expression of pheromone-responsive genes by mechanisms distinct from those used by the negative transcriptional regulators Cdc36, Cdc39, and Mot2. First, a Mot3-lexA fusion functions as a transcriptional activator. Second, Mot3 is a dose-dependent activator of several genes unrelated to pheromone response, including CYC1, SUC2, and LEU2. Third, insertion of consensus Mot3 binding sites (C/A/T)AGG(T/C)A activates a promoter in a MOT3-dependent manner. These findings, and the fact that consensus binding sites are found in the 5' flanking regions of many yeast genes, suggest that Mot3 is a globally acting transcriptional regulator. We hypothesize that Mot3 regulates expression of factors that attenuate signaling by the pheromone response pathway.
在酿酒酵母中,交配信息素反应由G蛋白和丝裂原活化蛋白(MAP)激酶依赖性信号通路的激活引发,并通过多种促进适应性或脱敏作用的机制而减弱。为了鉴定其产物对信息素信号传导起负调控作用的基因,我们筛选了可抑制过表达信号缺陷型G蛋白β亚基的野生型细胞的超适应性表型的突变。这鉴定出了MOT3中的隐性突变,该基因编码一种具有两个Cys2-His2锌指的核蛋白。发现Mot3是信息素反应和信息素诱导基因表达的剂量依赖性抑制剂,并且需要完整的信号通路来发挥其作用。几个结果表明,Mot3通过与负转录调节因子Cdc36、Cdc39和Mot2不同的机制减弱信息素反应性基因的表达。首先,Mot3-lexA融合蛋白起转录激活剂的作用。其次,Mot3是几个与信息素反应无关的基因(包括CYC1、SUC2和LEU2)的剂量依赖性激活剂。第三,共有Mot3结合位点(C/A/T)AGG(T/C)A的插入以MOT3依赖性方式激活启动子。这些发现,以及在许多酵母基因的5'侧翼区域发现共有结合位点这一事实,表明Mot3是一种全局作用的转录调节因子。我们推测Mot3调节减弱信息素反应途径信号传导的因子的表达。