Nikawa Jun-Ichi, Kamiuto Junko
Department of Bioscience and Bioinformatics, Faculty of Computer Science and Systems Engineering, Kyushu Institute of Technology, Iizuka, Fukuoka 820-8502, Japan.
J Biosci Bioeng. 2004;97(6):369-73. doi: 10.1016/S1389-1723(04)70220-0.
In Saccharomyces cerevisiae, the expression of several genes encoding enzymes involved in lipid metabolism is regulated by inositol and choline. The transcriptional heterodimeric complex composed of the gene products of INO2 and INO4 binds to a conserved cis-acting upstream activating sequence designated as the inositol-choline responsive element (ICRE), and activates the expression of these genes. In the presence of inositol and choline, the expression of these genes is downregulated and a functional OPI1 gene product is necessary for this repression. The promoter region of OPI1 contains one copy of ICRE, and here we analyzed the involvement of ICRE in the inositol-choline-mediated gene regulation of OPI1. Deletion analysis of the OPI1 promoter region, disruption of ICRE in it, and its activity in ino2- and ino4-disrupted strains showed that ICRE is essential for the expression of OPI1, and that the expression of OPI1 is dependent on the INO2 and INO4 gene products. Disruption of OPI1 resulted in the derepressed expression of OPI1 itself and no response to inositol-choline, showing that OPI1 is regulated in the same manner as the phospholipid biosynthetic genes. These results revealed the regulatory circuit of the expression of the positive regulatory gene INO2 and the negative regulatory gene OPI1.
在酿酒酵母中,几个编码参与脂质代谢的酶的基因的表达受肌醇和胆碱的调控。由INO2和INO4的基因产物组成的转录异二聚体复合物与一个保守的顺式作用上游激活序列(称为肌醇 - 胆碱反应元件,ICRE)结合,并激活这些基因的表达。在有肌醇和胆碱存在的情况下,这些基因的表达下调,并且功能性的OPI1基因产物对于这种抑制是必需的。OPI1的启动子区域包含一个ICRE拷贝,在此我们分析了ICRE在肌醇 - 胆碱介导的OPI1基因调控中的作用。对OPI1启动子区域的缺失分析、其中ICRE的破坏以及其在ino2和ino4缺失菌株中的活性表明,ICRE对于OPI1的表达是必需的,并且OPI1的表达依赖于INO2和INO4的基因产物。OPI1的破坏导致OPI1自身的去抑制表达,并且对肌醇 - 胆碱无反应,表明OPI1与磷脂生物合成基因以相同的方式被调控。这些结果揭示了正调控基因INO2和负调控基因OPI1表达的调控回路。