Tornow J, Santangelo G M
Biology Department, Portland State University, OR 97201-0751.
Gene. 1990 May 31;90(1):79-85. doi: 10.1016/0378-1119(90)90441-s.
The glycolytic form of alcohol dehydrogenase (ADHI) is encoded by the ADH1 gene of Saccharomyces cerevisiae. We found that efficient expression of the ADH1 gene requires a sequence between bp -635 and -615 with respect to the +1 mRNA start point; removal of this sequence reduced ADH1 mRNA levels 25-fold but did not affect carbon-source regulation. DNaseI footprinting analysis of the ADH1 promoter revealed the specific protection of a perfect match to UASRPG at -630 to -615. UASRPG is thought to be responsible for activation of transcription, via binding of the translation upstream factor (TUF), of genes encoding components of the translational apparatus. In band retardation assays, the promoters for the elongation factor 1 alpha-encoding genes (TEF1 and TEF2) competed for binding of the protein to the copy of UASRPG in the ADH1 promoter. We conclude that TUF is probably involved in activation of the bulk of ADH1 transcription. Further, we propose that TUF has a role in the activation of many or most glycolytic genes. If so, it is essential for efficient expression of a wide variety of functionally disparate products that are required by yeast cells for rapid growth.
酒精脱氢酶(ADHI)的糖酵解形式由酿酒酵母的ADH1基因编码。我们发现,ADH1基因的高效表达需要相对于+1 mRNA起始点在bp -635至-615之间的一段序列;去除该序列会使ADH1 mRNA水平降低25倍,但不影响碳源调控。对ADH1启动子的DNaseI足迹分析显示,在-630至-615处与UASRPG完全匹配的序列受到特异性保护。UASRPG被认为通过翻译上游因子(TUF)的结合来激活编码翻译装置组件的基因的转录。在凝胶阻滞试验中,编码延伸因子1α的基因(TEF1和TEF2)的启动子竞争该蛋白与ADH1启动子中UASRPG拷贝的结合。我们得出结论,TUF可能参与了大部分ADH1转录的激活。此外,我们提出TUF在许多或大多数糖酵解基因的激活中起作用。如果是这样,它对于酵母细胞快速生长所需的多种功能不同的产物的高效表达至关重要。