Kunoh T, Kaneko Y, Harashima S
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Yeast. 2000 Mar 30;16(5):439-49. doi: 10.1002/(SICI)1097-0061(20000330)16:5<439::AID-YEA536>3.0.CO;2-M.
The IME1 gene is essential for initiation of meiosis in the yeast Saccharomyces cerevisiae. Transcription of IME1 is detected under conditions of starvation for nitrogen and glucose, and in the presence of the MATa1 and MATalpha2 gene products. In our previous work, we have shown that there are two elements acting as TUP1-dependent upstream repression sequence (URS) and tup1 mutation-dependent upstream activation sequence (UAS) between nt -915 and -621 of the IME1 promoter under nutritional conditions. The region from -915 to -621 has also been reported to harbour meiotic URS and UAS when a/alpha cells were transferred to sporulation conditions. To identify proteins that are able to bind to the region, we screened a cDNA library fused with the Gal4-activation domain by means of the one-hybrid system. We identified a previously unknown gene (YDR451c), which we designated YHP1, encoding a homeodomain protein of the Drosophila antennapedia type. The region for binding of Yhp1 was delimited to the 28 bp region between nt -702 and -675 of the IME1 promoter in vivo and in vitro, and the 28 bp region harboured a URS activity in a Yhp1-dependent manner under nutrient growth conditions. Although a yhp1 single-disruption mutation did not give rise to a scorable phenotype under nutritional and sporulation conditions, the level of the YHP1 transcript was significantly lower in the cells grown in acetate medium (presporulation medium) and sporulation medium than those grown in glucose medium, and the reduction of YHP1 transcription in acetate medium coincided with an increment of the IME1 transcript. We suggest that the homeoprotein Yhp1 that binds directly to the 28 bp region of the IME1 promoter is a new repressor acting under glucose growth conditions.
IME1基因对于酿酒酵母中减数分裂的起始至关重要。IME1的转录在氮和葡萄糖饥饿条件下以及存在MATa1和MATalpha2基因产物时被检测到。在我们之前的工作中,我们已经表明,在营养条件下,IME1启动子的-915至-621核苷酸之间存在两个元件,分别作为TUP1依赖性上游抑制序列(URS)和tup1突变依赖性上游激活序列(UAS)。当a/α细胞转移到孢子形成条件下时,-915至-621区域也被报道含有减数分裂URS和UAS。为了鉴定能够结合该区域的蛋白质,我们通过单杂交系统筛选了与Gal4激活域融合的cDNA文库。我们鉴定出一个以前未知的基因(YDR451c),我们将其命名为YHP1,它编码果蝇触角足类型的同源域蛋白。在体内和体外,Yhp1的结合区域被限定在IME1启动子的-702至-675核苷酸之间的28 bp区域,并且在营养生长条件下,该28 bp区域以Yhp1依赖性方式具有URS活性。尽管yhp1单破坏突变在营养和孢子形成条件下没有产生可评分的表型,但在乙酸盐培养基(孢子形成前培养基)和孢子形成培养基中生长的细胞中YHP1转录本的水平明显低于在葡萄糖培养基中生长的细胞,并且乙酸盐培养基中YHP1转录的减少与IME1转录本的增加相一致。我们认为直接结合IME1启动子28 bp区域的同源蛋白Yhp1是在葡萄糖生长条件下起作用的一种新的阻遏物。