Theodoris G, Bisson L F
Department of Enology and Viticulture, University of California, One Shields Avenue, Davis, CA 95616-8749, USA.
FEMS Microbiol Lett. 2001 Apr 1;197(1):73-7. doi: 10.1111/j.1574-6968.2001.tb10585.x.
Mutations in the yeast SNF3 gene affect glucose sensing and snf3 mutants show defective growth on glucose. DNA sequence dependent suppressing elements (DDSEs) are regions located in the promoters of yeast glucose transporter (HXT) genes that when present in high copy suppress the snf3 growth defect. Here we provide evidence that the multicopy DDSE suppression is due to the titration of the Rgt1p transcriptional repressor. The DDSE region from HXT4 was found to function as a UAS sequence rendering a UAS(gal)-less LacZ gene fused to the GAL1 promoter responsive to glucose induction. Expression mediated by the UAS(DDSE) was dependent upon the presence of Snf3p. Expression was elevated to a high level in an rgt1 mutant in the absence of Snf3p suggesting that this DDSE region contains binding sites for the Rgt1p transcriptional repressor/activator. The UAS(DDSE) led to expression in a grr1 mutant background, which confers a defect in inactivation of Rgt1p, as predicted from the model. The presence of tandem repeats of the putative Rgt1p binding site gave results similar to those of the DDSE, suggesting that loss of repression is due to the presence of Rgt1p footprint in the multicopy DDSE.
酵母SNF3基因的突变会影响葡萄糖感应,且snf3突变体在葡萄糖上生长存在缺陷。DNA序列依赖性抑制元件(DDSEs)是位于酵母葡萄糖转运蛋白(HXT)基因启动子中的区域,当以高拷贝形式存在时可抑制snf3的生长缺陷。在此我们提供证据表明多拷贝DDSE抑制是由于Rgt1p转录阻遏物的滴定作用。发现来自HXT4的DDSE区域可作为一个上游激活序列(UAS)发挥作用,使与GAL1启动子融合的无UAS(gal)的LacZ基因对葡萄糖诱导产生反应。由UAS(DDSE)介导的表达依赖于Snf3p的存在。在无Snf3p的情况下,在rgt1突变体中表达升高至高水平,这表明该DDSE区域含有Rgt1p转录阻遏物/激活物的结合位点。正如模型所预测的,UAS(DDSE)在grr1突变体背景中导致表达,该突变体在Rgt1p失活方面存在缺陷。推测的Rgt1p结合位点的串联重复序列的存在产生了与DDSE相似的结果,表明抑制作用的丧失是由于多拷贝DDSE中存在Rgt1p足迹。