Rüth J, Hirt H, Schweyen R J
Institute of Microbiology and Genetics, University of Vienna, Austria.
Mol Gen Genet. 1992 Nov;235(2-3):365-72. doi: 10.1007/BF00279382.
The cauliflower mosaic virus 35S promoter confers strong gene expression in plants, animals and fission yeast, but not in budding yeast. On investigating this paradox, we found that in budding yeast the promoter acts through two domains. Whereas the upstream domain acts as a silencer, the downstream domain couples expression to the nutritional state of the cells via the RAS/cAMP pathway. Point mutations indicate that two boxes with similarity to the cAMP regulated element (CRE) of mammalian cells mediate this response. Gel retardation assays show that, in both yeast and plant protein extracts, factors bind to this promoter element. Therefore, transcriptional activation appears to be highly conserved at the level of transcription factors and specific DNA target elements in eukaryotes. This offers new ways to investigate gene regulation mechanisms of higher eukaryotes, which are not as amenable to genetic analysis as yeast.
花椰菜花叶病毒35S启动子在植物、动物和裂殖酵母中能赋予强基因表达,但在芽殖酵母中却不能。在研究这一矛盾现象时,我们发现,在芽殖酵母中该启动子通过两个结构域发挥作用。上游结构域起沉默子的作用,而下游结构域则通过RAS/环磷酸腺苷(cAMP)途径将表达与细胞的营养状态联系起来。点突变表明,两个与哺乳动物细胞的cAMP调节元件(CRE)相似的框介导了这种反应。凝胶阻滞试验表明,在酵母和植物蛋白提取物中,都有因子与该启动子元件结合。因此,转录激活在真核生物的转录因子和特定DNA靶元件水平上似乎高度保守。这为研究高等真核生物的基因调控机制提供了新方法,高等真核生物不像酵母那样易于进行遗传分析。