Horecka J, Sprague G F
Institute of Molecular Biology, University of Oregon, Eugene 97403-1229, USA.
Genetics. 1996 Nov;144(3):905-21. doi: 10.1093/genetics/144.3.905.
In haploid Saccharomyces cerevisiae cells, mating pheromones activate a signal transduction pathway that leads to cell cycle arrest in the G1 phase and to transcription induction of genes that promote conjugation. To identify genes that link the signal transduction pathway and the cell cycle machinery, we developed a selection strategy to isolate yeast mutants specifically defective for G1 arrest. Several of these mutants identified previously known genes, including CLN3, FUS3, and FAR1. In addition, a new gene, FAR3, was identified and characterized. FAR3 encodes a novel protein of 204 amino acid residues that is dispensable for viability. Northern blot experiments indicated that FAR3 expression is constitutive with respect to cell type, pheromone treatment, and cell cycle position. As a first step toward elucidating the mechanism by which Far3 promotes pheromone-mediated G1 arrest, we performed genetic and molecular experiments to test the possibility that Far3 participates in one of the heretofore characterized mechanisms, namely Fus3/Far1-mediated inhibition of Cdc28-Cln kinase activity, G1 cyclin gene repression, and G1 cyclin protein turnover. Our data indicate that Far3 effects G1 arrest by a mechanism distinct from those previously known.
在单倍体酿酒酵母细胞中,交配信息素激活一条信号转导通路,该通路导致细胞周期在G1期停滞,并诱导促进接合的基因转录。为了鉴定连接信号转导通路和细胞周期机制的基因,我们开发了一种筛选策略,以分离出在G1期停滞方面存在特异性缺陷的酵母突变体。其中一些突变体鉴定出了先前已知的基因,包括CLN3、FUS3和FAR1。此外,还鉴定并表征了一个新基因FAR3。FAR3编码一种由204个氨基酸残基组成的新型蛋白质,该蛋白质对于细胞生存力是可有可无的。Northern印迹实验表明,FAR3的表达在细胞类型、信息素处理和细胞周期位置方面是组成型的。作为阐明Far3促进信息素介导的G1期停滞机制的第一步,我们进行了遗传和分子实验,以测试Far3是否参与了迄今为止已表征的机制之一,即Fus3/Far1介导的对Cdc28-Cln激酶活性的抑制、G1期细胞周期蛋白基因的抑制以及G1期细胞周期蛋白的周转。我们的数据表明,Far3通过一种不同于先前已知机制的方式影响G1期停滞。