Kunoh T, Kaneko Y, Harashima S
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Biochem Biophys Res Commun. 2000 Nov 19;278(2):344-8. doi: 10.1006/bbrc.2000.3800.
The IME1 gene is essential for initiation of meiosis in Saccharomyces cerevisiae. Transcription of IME1 is induced under starvation for nitrogen and glucose and in the presence of the MATa1 and MATalpha2 gene products. We have shown in our previous work that homeoprotein Yhp1 binds to a 28-bp region between nt -702 and -675 of the IME1 promoter in vivo and in vitro. We also revealed that the 28-bp region fused with a reporter gene harbored Yhp1-dependent URS (upstream repression sequence) activity and that the transcription of YHP1 was repressed by nonfermentable carbon source. In this study, we found, using a 5'-deletion series of the YHP1 promoter fused with a reporter gene, that the URS responsible for repression of YHP1 transcription with a nonfermentable carbon source is located at a region from nt -696 to -466 of the YHP1 promoter. We also identified and delimited a UAS (upstream activation sequence), which confers activation in both fermentable and nonfermentable carbon source media, to be from nt -356 to -306 of the YHP1 promoter. The UAS of the YHP1 promoter contained an MCE (Mcm1 control element) that is a target of the general transcription factor Mcm1 and is known to be involved in positive regulation by the two-component regulator Sln1. Consistent with this fact, the YHP1 transcription level was reduced in the Deltasln1 mutant, indicating that the two-component regulator Sln1 is involved in activation of YHP1 transcription.
IME1基因对于酿酒酵母减数分裂的起始至关重要。IME1的转录在氮源和葡萄糖饥饿条件下以及在MATa1和MATalpha2基因产物存在时被诱导。我们在先前的工作中表明,同源蛋白Yhp1在体内和体外均与IME1启动子的-702至-675核苷酸之间的28碱基对区域结合。我们还发现,与报告基因融合的28碱基对区域具有Yhp1依赖性的URS(上游抑制序列)活性,并且YHP1的转录受到不可发酵碳源的抑制。在本研究中,我们使用与报告基因融合的YHP1启动子的5'-缺失系列发现,负责在不可发酵碳源下抑制YHP1转录的URS位于YHP1启动子的-696至-466核苷酸区域。我们还鉴定并界定了一个UAS(上游激活序列),其在可发酵和不可发酵碳源培养基中均赋予激活作用,位于YHP1启动子的-356至-306核苷酸处。YHP1启动子的UAS包含一个MCE(Mcm1控制元件),它是一般转录因子Mcm1的靶点,并且已知参与双组分调节因子Sln1的正调控。与此事实一致,在Deltasln1突变体中YHP1转录水平降低,表明双组分调节因子Sln1参与YHP1转录的激活。