Nishizawa M, Suzuki Y, Nogi Y, Matsumoto K, Fukasawa T
Department of Microbiology, Keio University School of Medicine, Tokyo, Japan.
Proc Natl Acad Sci U S A. 1990 Jul;87(14):5373-7. doi: 10.1073/pnas.87.14.5373.
GAL11 was first identified as a gene required for full expression of some of the galactose-inducible genes in the yeast Saccharomyces cerevisiae. A null mutation within the GAL11 locus causes defects in mating, growth on nonfermentable carbon sources, and sporulation of gal11 homozygotes. The mating defect was observed only in MAT alpha gal11 strains. Northern hybridization analysis revealed that a gal11 mutation impaired transcription of alpha-specific genes (MF alpha 1 and STE3) but not of an a-specific gene (STE2). Furthermore, this mutation reduced expression of the MAT alpha locus, suggesting that a deficiency in MAT alpha 1 protein is responsible for the reduced expression of alpha-specific genes. Since general regulatory factor I (GRFI)/repressor/activator site binding protein 1 (RAP1)/translation upstream factor (TUF) is believed to be an activator of MAT alpha expression, we examined whether PYK1, which is known to be regulated by GRFI/RAP1/TUF, is also affected by the gal11 mutation. It was determined that the level of PYK1 message was significantly lowered by the mutation. The requirement for functional GAL11 in transcriptional activation was bypassed when either the upstream activating sequence of galactose-inducible genes or of PYK1 was placed very close to the TATA box, suggesting that one of the Gal11 protein functions is to mediate the activation signal of Gal4 and GRFI/RAP1/TUF, when the respective binding site is situated at the naturally occurring distance from the TATA box.
GAL11最初被鉴定为酿酒酵母中一些半乳糖诱导基因完全表达所必需的基因。GAL11基因座内的无效突变会导致交配缺陷、在非发酵碳源上生长以及gal11纯合子的孢子形成缺陷。仅在MATα gal11菌株中观察到交配缺陷。Northern杂交分析显示,gal11突变会损害α特异性基因(MFα1和STE3)的转录,但不会损害a特异性基因(STE2)的转录。此外,该突变降低了MATα基因座的表达,这表明MATα1蛋白的缺乏是α特异性基因表达降低的原因。由于一般调节因子I(GRFI)/阻遏物/激活物位点结合蛋白1(RAP1)/翻译上游因子(TUF)被认为是MATα表达的激活剂,我们研究了已知受GRFI/RAP1/TUF调节的PYK1是否也受gal11突变的影响。结果确定该突变显著降低了PYK1信使水平。当半乳糖诱导基因或PYK1的上游激活序列非常靠近TATA框时,转录激活中对功能性GAL11的需求被绕过,这表明当各自的结合位点位于距TATA框的自然距离时,Gal11蛋白的功能之一是介导Gal4和GRFI/RAP1/TUF的激活信号。