Chambers A, Tsang J S, Stanway C, Kingsman A J, Kingsman S M
Department of Biochemistry, University of Oxford, United Kingdom.
Mol Cell Biol. 1989 Dec;9(12):5516-24. doi: 10.1128/mcb.9.12.5516-5524.1989.
The promoter of the yeast glycolytic gene encoding phosphoglycerate kinase (PGK) contains an upstream activation sequence between bases -538 and -402 upstream of the initiating ATG. The upstream activation sequence contains multiple functional elements, including an essential region called the activator core (AC) sequence and three copies of the pentamer 5'-CTTCC-3'. The AC sequence shows strong homology to the consensus binding sites for the yeast proteins RAP1 (GRF1) and TUF. We have demonstrated that the yeast protein which interacts with the AC sequence is the DNA-binding protein RAP1. Expression of the PGK gene is found to be regulated according to the carbon source in the growth medium. PGK mRNA levels are high in yeast cells grown in glucose medium but low in yeast cells grown in media containing carbon sources such as pyruvate and acetate. This carbon source regulation of transcription was found to be mediated, in part, via regulation of RAP1 binding to the AC sequence. The promoters of many other yeast glycolytic genes also contain consensus RAP1-binding sites and copies of the CTTCC pentamer. This suggests that RAP1 may be involved in transcriptional control of many other glycolytic genes in addition to the PGK gene.
编码磷酸甘油酸激酶(PGK)的酵母糖酵解基因的启动子在起始ATG上游-538至-402碱基之间含有一个上游激活序列。该上游激活序列包含多个功能元件,包括一个称为激活剂核心(AC)序列的必需区域和三个五聚体5'-CTTCC-3'的拷贝。AC序列与酵母蛋白RAP1(GRF1)和TUF的共有结合位点具有很强的同源性。我们已经证明,与AC序列相互作用的酵母蛋白是DNA结合蛋白RAP1。发现PGK基因的表达根据生长培养基中的碳源进行调节。在葡萄糖培养基中生长的酵母细胞中PGK mRNA水平较高,但在含有丙酮酸和乙酸盐等碳源的培养基中生长的酵母细胞中水平较低。发现这种转录的碳源调节部分是通过调节RAP1与AC序列的结合来介导的。许多其他酵母糖酵解基因的启动子也含有共有RAP1结合位点和CTTCC五聚体的拷贝。这表明RAP1可能除了参与PGK基因的转录控制外,还参与许多其他糖酵解基因的转录控制。