Amakasu H, Suzuki Y, Nishizawa M, Fukasawa T
Department of Microbiology, Keio University School of Medicine, Tokyo, Japan.
Genetics. 1993 Jul;134(3):675-83. doi: 10.1093/genetics/134.3.675.
Recessive mutations of GAL11 in Saccharomyces cerevisiae cause pleiotropic defects that include weak fermentation of galactose, alpha-specific sterility and slow growth on nonfermentable carbon sources. Recent experiments suggest that Gal11P functions as a "co-activator" that links transcriptional activators, such as Gal4p and Grf1p/Rap1p/Tuf1p, with the basic transcription machinery. In the present experiments we isolated a gene, SGE1, that suppresses gal11 for growth on ethidium bromide/galactose agar when the gene was present in two or more copies. The other gal11 phenotypes were not suppressed by SGE1 in the multiple-copy state. Multiple copies of SGE1 increased expression of galactose-inducible genes in gal11 yeast, presumably at the level of transcription. When SGE1 was disrupted in wild-type yeast, the expression of galactose-inducible genes decreased to 50-60% of the wild-type level, presumably due to effect on transcription. Complete DNA sequence analysis revealed that SGE1 encodes a predicted protein of 543 amino acids. SGE1-specific mRNA of 1.8 kilonucleotides was detected by Northern analysis along the direction of the open reading frame. The gene mapped near RAD56, at the right end of chromosome 16.
酿酒酵母中GAL11的隐性突变会导致多效性缺陷,包括半乳糖发酵能力弱、α特异性不育以及在非发酵性碳源上生长缓慢。最近的实验表明,Gal11P作为一种“共激活因子”,将转录激活因子,如Gal4p和Grf1p/Rap1p/Tuf1p,与基本转录机制联系起来。在本实验中,我们分离出一个基因SGE1,当该基因以两个或更多拷贝存在时,它能在溴化乙锭/半乳糖琼脂上抑制gal11的生长。SGE1在多拷贝状态下不会抑制其他gal11表型。SGE1的多个拷贝增加了gal11酵母中半乳糖诱导基因的表达,推测是在转录水平上。当SGE1在野生型酵母中被破坏时,半乳糖诱导基因的表达下降到野生型水平的50 - 60%,推测是由于对转录的影响。完整的DNA序列分析表明,SGE1编码一个预测的由543个氨基酸组成的蛋白质。通过Northern分析沿着开放阅读框的方向检测到1.8千核苷酸的SGE1特异性mRNA。该基因定位于16号染色体右端靠近RAD56的位置。