Mukai Y, Harashima S, Oshima Y
Department of Biotechnology, Faculty of Engineering, Osaka University, Japan.
Mol Cell Biol. 1991 Jul;11(7):3773-9. doi: 10.1128/mcb.11.7.3773-3779.1991.
We have cloned a DNA fragment complementing the aar1 mutation defective in the a1-alpha 2 repression of the alpha 1 cistron and haploid-specific genes in Saccharomyces cerevisiae. Nucleotide sequence and mapping data indicated that the AAR1 gene is identical with TUP1, which is allelic to the SFL2, FLK1, CYC9, UMR7, AMM1, and AER2 genes, whose mutations are known to confer a variety of phenotypes, such as thymidine uptake, flocculation, insensitivity to glucose repression, a defect in UV-induced mutagenesis, and a defect in ARS plasmid maintenance. The TUP1/AER2 protein is known to have significant similarity with the beta subunits of G proteins in the C-terminal half, in two glutamine-rich domains in the N-terminal half, and in a central region rich in serine and threonine residues. Disruption of the chromosomal AAR1 gene in alpha and a/alpha cells conferred the nonmating phenotype, and the a/alpha diploids could not sporulate. The AAR1/TUP1 gene is transcribed into a 2.5-kb mRNA independently of the mating-type information of the cell. These observations and mRNA analysis of cell-type-specific genes indicated that the AAR1/TUP1 protein is also indispensable for a1-alpha 2 repression of RME1 and for alpha 2 repression of a-specific genes.
我们克隆了一个DNA片段,它能互补酿酒酵母中α1顺反子和单倍体特异性基因的α1-α2阻遏缺陷型aar1突变。核苷酸序列和定位数据表明,AAR1基因与TUP1相同,TUP1与SFL2、FLK1、CYC9、UMR7、AMM1和AER2基因等位,已知这些基因的突变会导致多种表型,如胸苷摄取、絮凝、对葡萄糖阻遏不敏感、紫外线诱导诱变缺陷以及ARS质粒维持缺陷。已知TUP1/AER2蛋白在C端的一半与G蛋白的β亚基有显著相似性,在N端的两个富含谷氨酰胺的结构域以及富含丝氨酸和苏氨酸残基的中央区域也有相似性。α细胞和a/α细胞中染色体AAR1基因的破坏导致了不交配表型,并且a/α二倍体不能形成孢子。AAR1/TUP1基因转录成一个2.5kb的mRNA,与细胞的交配型信息无关。这些观察结果以及对细胞类型特异性基因的mRNA分析表明,AAR1/TUP1蛋白对于RME1的α1-α2阻遏以及a特异性基因的α2阻遏也是必不可少的。