Kos W, Kal A J, van Wilpe S, Tabak H F
E.C. Slater Institute, Academic Medical Center, University of Amsterdam, The Netherlands.
Biochim Biophys Acta. 1995 Oct 17;1264(1):79-86. doi: 10.1016/0167-4781(95)00127-3.
In Saccharomyces cerevisiae induction of the FOX3 gene, encoding peroxisomal 3-oxoacyl-CoA thiolase, by growth on oleate as sole carbon source, is exerted via the cis-acting DNA element designated oleate response element (ORE) (Einerhand et al. (1991) Eur. J. Biochem. 200, 113-122). The transcription factor(s) binding to this upstream activation site (UAS) are still unknown, however. Induction of another peroxisomal enzyme, citrate synthase (CIT2) is dependent on the products of two genes called RTG1 and RTG2 (Liao and Butow (1993) Cell 72, 61-71). In the present study we have investigated whether RTG1 controls other genes coding for peroxisomal proteins, and whether such control takes place via the ORE. A number of genes coding for a variety of peroxisomal proteins such as: thiolase and catalase (peroxisomal matrix proteins), PAS3p (a peroxisomal membrane protein) and PAS10p (a protein involved in the import of peroxisomal proteins) were studied in their response to RTG1. Although the RTG1 and 2 products proved to be required for the increase in number and volume of peroxisomes upon induction by oleate, the single promoter output of the chosen set of genes remained practically unchanged in a rtg1 mutant strain. In addition gel retardation experiments indicated that RTG1 does not bind to the ORE. The behavior of genes coding for the various proteins also varied during repression, derepression and induction, indicating that probably a number of proteins are involved in tuning the output of each gene to cellular demand.
在酿酒酵母中,以油酸作为唯一碳源生长时,编码过氧化物酶体3-氧代酰基辅酶A硫解酶的FOX3基因的诱导是通过称为油酸反应元件(ORE)的顺式作用DNA元件实现的(艾纳汉德等人,(1991年)《欧洲生物化学杂志》200,113 - 122)。然而,与这个上游激活位点(UAS)结合的转录因子仍然未知。另一种过氧化物酶体酶柠檬酸合酶(CIT2)的诱导依赖于两个名为RTG1和RTG2的基因的产物(廖和布托,(1993年)《细胞》72,61 - 71)。在本研究中,我们调查了RTG1是否控制其他编码过氧化物酶体蛋白的基因,以及这种控制是否通过ORE发生。研究了许多编码各种过氧化物酶体蛋白的基因,如:硫解酶和过氧化氢酶(过氧化物酶体基质蛋白)、PAS3p(一种过氧化物酶体膜蛋白)和PAS10p(一种参与过氧化物酶体蛋白导入的蛋白)对RTG1的反应。尽管RTG1和2的产物被证明是油酸诱导后过氧化物酶体数量和体积增加所必需的,但在rtg1突变株中,所选基因集的单个启动子输出实际上保持不变。此外,凝胶阻滞实验表明RTG1不与ORE结合。编码各种蛋白质的基因在阻遏、去阻遏和诱导过程中的行为也有所不同,这表明可能有许多蛋白质参与将每个基因的输出调整到细胞需求。