Mukai Y, Harashima S, Oshima Y
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Mol Cell Biol. 1993 Apr;13(4):2050-60. doi: 10.1128/mcb.13.4.2050-2060.1993.
Sterile mutants of Saccharomyces cerevisiae were isolated from alpha * cells having the a/alpha aar1-6 genotype (exhibiting alpha mating ability and weak a mating ability as a result of a defect in a1-alpha 2 repression). Among these sterile mutants, we found two ste5 mutants together with putative ste7, ste11, and ste12 mutants of the signal transduction pathway of mating pheromones. The amino acid sequence of the Ste5p protein predicted from the nucleotide sequence of a cloned STE5 DNA has a domain rich in acidic amino acids close to its C terminus, a cysteine-rich sequence, resembling part of a zinc finger structure, in its N-terminal half, and a possible target site of cyclic AMP-dependent protein kinase at its C terminus. Northern (RNA) blot analysis revealed that STE5 transcription is under a1-alpha 2-Aar1p repression. The MAT alpha 1 cistron has a single copy of the pheromone response element in its 5' upstream region, and its basal level of transcription was reduced in these ste mutant cells. However, expression of the MAT alpha 1 cistron was not enhanced appreciably by pheromone signals. One of the ste5 mutant alleles conferred a sterile phenotype to a/alpha aar1-6 cells but a mating ability to MATa cells.
从具有a/α aar1-6基因型(由于a1-α2抑制缺陷而表现出α交配能力和较弱的a交配能力)的α*细胞中分离出酿酒酵母的无菌突变体。在这些无菌突变体中,我们发现了两个ste5突变体以及交配信息素信号转导途径中假定的ste7、ste11和ste12突变体。从克隆的STE5 DNA的核苷酸序列预测的Ste5p蛋白的氨基酸序列在其C末端附近有一个富含酸性氨基酸的结构域,在其N端一半有一个富含半胱氨酸的序列,类似于锌指结构的一部分,并且在其C末端有一个可能的环磷酸腺苷依赖性蛋白激酶的靶位点。Northern(RNA)印迹分析表明,STE5转录受a1-α2-Aar1p抑制。MATα1顺反子在其5'上游区域有一个单一拷贝的信息素反应元件,并且在这些ste突变体细胞中其基础转录水平降低。然而,信息素信号并未明显增强MATα1顺反子的表达。其中一个ste5突变等位基因赋予a/α aar1-6细胞无菌表型,但赋予MATa细胞交配能力。