Palomino Aaron, Herrero Pilar, Moreno Fernando
Departamento de Bioquímica y Biología Molecular, Universidad de Oviedo, Campus del Cristo, Edificio Santiago Gascón, 33006 Oviedo, Spain.
Biochem J. 2005 Jun 1;388(Pt 2):697-703. doi: 10.1042/BJ20050160.
Expression of HXK2, a gene encoding a Saccharomyces cerevisiae bifunctional protein with catalytic and regulatory functions, is controlled by glucose availability, being activated in the presence of glucose and inhibited when the levels of the sugar are low. In the present study, we identified Rgt1 as a transcription factor that, together with the Med8 protein, is essential for repression of the HXK2 gene in the absence of glucose. Rgt1 represses HXK2 expression by binding specifically to the motif (CGGAAAA) located at -395 bp relative to the ATG translation start codon in the HXK2 promoter. Disruption of the RGT1 gene causes an 18-fold increase in the level of HXK2 transcript in the absence of glucose. Rgt1 binds to the RGT1 element of HXK2 promoter in a glucose-dependent manner, and the repression of target gene depends on binding of Rgt1 to DNA. The physiological significance of the connection between two glucose-signalling pathways, the Snf3/Rgt2 that causes glucose induction and the Mig1/Hxk2 that causes glucose repression, was also analysed.
HXK2是一种编码具有催化和调节功能的酿酒酵母双功能蛋白的基因,其表达受葡萄糖可用性的控制,在葡萄糖存在时被激活,而当糖水平较低时被抑制。在本研究中,我们鉴定出Rgt1作为一种转录因子,它与Med8蛋白一起,在缺乏葡萄糖时对HXK2基因的抑制至关重要。Rgt1通过特异性结合HXK2启动子中相对于ATG翻译起始密码子位于-395 bp处的基序(CGGAAAA)来抑制HXK2的表达。在缺乏葡萄糖的情况下,RGT1基因的破坏导致HXK2转录水平增加18倍。Rgt1以葡萄糖依赖的方式与HXK2启动子的RGT1元件结合,并且靶基因的抑制取决于Rgt1与DNA的结合。我们还分析了两条葡萄糖信号通路之间联系的生理意义,即引起葡萄糖诱导的Snf3/Rgt2和引起葡萄糖抑制的Mig1/Hxk2。