Shen H, Dowhan W
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston, Texas 77225, USA.
J Biol Chem. 1998 May 8;273(19):11638-42. doi: 10.1074/jbc.273.19.11638.
The PGS1 gene of Saccharomyces cerevisiae encodes phosphatidylglycerophosphate (PG-P) synthase. PG-P synthase activity is regulated by factors affecting mitochondrial development and through cross-pathway control by inositol. The molecular mechanism of this regulation was examined by using a reporter gene under control of the PGS1 gene promoter (PPGS1-lacZ). Gene expression subject to carbon source regulation was monitored both at steady-state level and during the switch between different carbon sources. Cells grown in a non-fermentable carbon source had beta-galactosidase levels 3-fold higher than those grown in glucose. A shift from glucose to lactate rapidly raised the level of gene expression, whereas a shift back to glucose had the opposite effect. In either a pgs1 null mutant or a rho mutant grown in glucose, PPGS1-lacZ expression was 30-50% of the level in wild type cells. Addition of inositol to the growth medium resulted in a 2-3-fold reduction in gene expression in wild type cells. In ino2 and ino4 mutants, gene expression was greatly reduced and was not subject to inositol regulation consistent with inositol repression being dependent on the INO2 and INO4 regulatory genes. PPGS1-lacZ expression was elevated in a cds1 null mutant in the presence or absence of inositol, indicating that the capacity to synthesize CDP-diacylglycerol affects gene expression. Lack of cardiolipin synthesis (cls1 null mutant) had no effect on reporter gene expression.
酿酒酵母的PGS1基因编码磷脂酰甘油磷酸(PG-P)合酶。PG-P合酶活性受影响线粒体发育的因素调控,并通过肌醇进行跨途径控制。利用PGS1基因启动子(PPGS1-lacZ)控制下的报告基因研究了这种调控的分子机制。在稳态水平以及不同碳源转换期间监测受碳源调控的基因表达。在非发酵碳源中生长的细胞的β-半乳糖苷酶水平比在葡萄糖中生长的细胞高3倍。从葡萄糖转换为乳酸会迅速提高基因表达水平,而转换回葡萄糖则有相反的效果。在葡萄糖中生长的pgs1缺失突变体或rho突变体中,PPGS1-lacZ表达是野生型细胞中水平的30-50%。向生长培养基中添加肌醇会导致野生型细胞中的基因表达降低2-3倍。在ino2和ino4突变体中,基因表达大大降低且不受肌醇调控,这与肌醇阻遏依赖于INO2和INO4调控基因一致。在存在或不存在肌醇的情况下,cds1缺失突变体中的PPGS1-lacZ表达均升高,表明合成CDP-二酰基甘油的能力会影响基因表达。缺乏心磷脂合成(cls1缺失突变体)对报告基因表达没有影响。