Achstetter T
Institut für Botanik, Universität Regensburg, Federal Republic of Germany.
Mol Cell Biol. 1989 Oct;9(10):4507-14. doi: 10.1128/mcb.9.10.4507-4514.1989.
Production of the mating pheromone alpha-factor was examined in Saccharomyces cerevisiae MAT alpha cells that had been exposed to the mating pheromone a-factor. A 2-h treatment with a-factor caused a significant increase in alpha-factor concentration in the medium as demonstrated by a halo assay. MF alpha 1 is one of the two genes coding for a precursor of alpha-factor. A Northern (RNA) analysis of total RNA from a-factor-treated MAT alpha cells revealed a rapid two- to threefold increase in MF alpha 1 transcript levels, reaching maximum within 60 min of exposure to the pheromone. Pheromone induction did not require ongoing protein synthesis. a-Factor-induced MF alpha 1 expression was quantitated by analysis of an MF alpha 1::SUC2 fusion gene whose product was assayed for invertase activity. Expression of the MF alpha 1::SUC2 gene in MAT alpha cells responded to the a-factor signal like the chromosomal version of MF alpha 1. Maturation of the alpha-factor precursor involves three proteolytic activities which are encoded by the KEX1, KEX2, and STE13 genes, respectively. Two of these genes, namely, KEX2 and STE13, were examined for pheromone-induced expression. Only the STE13 gene exhibited pheromone induction at the transcriptional level.
在暴露于交配信息素a-因子的酿酒酵母MATα细胞中检测了交配信息素α-因子的产生。用a-因子处理2小时导致培养基中α-因子浓度显著增加,这通过晕圈试验得以证明。MFα1是编码α-因子前体的两个基因之一。对经a-因子处理的MATα细胞的总RNA进行的Northern(RNA)分析显示,MFα1转录水平迅速增加两到三倍,在暴露于信息素后60分钟内达到最大值。信息素诱导不需要持续的蛋白质合成。通过分析MFα1::SUC2融合基因来定量a-因子诱导的MFα1表达,其产物通过检测转化酶活性来测定。MATα细胞中MFα1::SUC2基因的表达对a-因子信号的反应与MFα1的染色体版本相似。α-因子前体的成熟涉及三种蛋白水解活性,分别由KEX1、KEX2和STE13基因编码。检测了其中两个基因,即KEX2和STE13的信息素诱导表达。只有STE13基因在转录水平上表现出信息素诱导。