Analysis of the signal transduction of the formation of subcellular organelles, peroxisomes

亚细胞器、过氧化物酶体形成的信号转导分析

• 批准号: 09660081
• 负责人: UEDA Mitsuyoshi
• 金额: $ 2.11万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660081/
• 关键词: Subcellular organelle; Peroxisomes; Signal transduction; Yeast; Control of gene expression; Homologous recombination; Cascade; Oleic acid

Candida tropicalis is an asporogenic diploid yeast which is characterized by its ability to utilize n-alkanes as a sole carbon and energy source. In assimilating n-alkanes, C.tropicalis shows a change in its intracellular structure by proliferating a large number of peroxisomes. Peroxisomes are versatile, single-membrane-bound organelles occurring in most of eukaryotic cells. Concomitant with peroxisome proliferation, the activities of peroxisomal enzymes are induced in n-alkane-grown cells. On the other hand, they are synthesized at low rate or not at all in glucose-grown cells. This phenomenon is called as glucose repression. Isociterate lyase (ICL), a key enzyme of the glyoxylate cycle being localized in peroxisomes, is under the control of glucose repression. Promoters of Saccharomyces cerevisiae controlled by the glucose repression mechanism were used to search for cis-regulatory elements responsible for derepression of gene expression. Various trans-acting factors which are invol … More ved in the genetic regulation of the glucose repression in S.cerevisiae, were also identified. The fact that UPR-ICL-mediated transcription of C.tropicalis is still glucose-repressive in S.cerevisiae demonstrates that similar mechanisms are commonly existing in both of the yeast strains concerning the regulation of ICL gene. A mutant was isolated that failed to derepress the UPR-ICL-mediated gene expression in acetate medium, and the gene (FILl) that complemented this mutation was isolated. The fill null mutant in which FILl is disrupted could not grow on acetate or ethanol, and the derepression of the isocitrate lyase encoded by ICLl in S.cerevisiae was also defected. The amino acid sequence of Fillp (230 amino acids) showed similarity to ribosome recycling factors (RRFs) of prokaryotes. Compared to prokaryotic RRFs, Fil ip had an N-terminal 46 amino acid extension which was shown to be able to function as a mitochondrial targeting sequence. The subcellular fractionation of the DELTAfil 1 strain showed that protein constituents of the mitochondrial fraction of the DELTAfil 1 strain differed from those of the wild type strain, but resembled those of chloramphenicol-treated cells or rho。 cells. These results suggest that Fillp is necessary for protein synthesis in mitochondria of S.cerevisiae. The results indicate the presence of a communication pathway between mitochondria and the nucleus which represses expression of genes encoding the key enzymes of the glyoxylate cycle and gluconeogenic pathway when there is a deficiency in the mitochondrial respiratory chain. Less

Tropicalis念珠菌是一种生成二倍体酵母，其特征在于其利用N-烷烃作为唯一的碳和能源的能力。在吸收N-烷烃时，C.tropicalis通过增殖大量的过氧化物体来显示其细胞内结构的变化。在大多数真核细胞中，过氧化物体是通用的单膜结合细胞器。与过氧化物组增殖的伴随，在N-烷烃生长的细胞中诱导过氧酶的活性。另一方面，它们在葡萄糖生长的细胞中以低速率或根本不合成。该现象称为葡萄糖表达。等素裂解酶（ICL）是葡萄糖表达的控制，是乙氧基循环的关键酶。由葡萄糖表达机制控制的酿酒酵母的启动子用于搜索负责消除基因表达的顺式调节元件。还鉴定出了涉及的各种涉及的跨作用因子。在塞维西亚链球菌中，C.tropicalis的UP-ICL介导的转录仍然是葡萄糖抑制，这表明，与ICL基因调节有关的两个酵母菌菌株中通常存在类似的机制。分离出一个突变体，未能解除乙酸盐培养基中UPR-ICL介导的基因表达，而完成该突变的基因（填充）是分离的。填充填充的填充零突变体无法在乙酸盐或乙醇上生长，并且还违反了由ICLL在Cerevisiae中编码的异位酸酯裂解酶的消除。 FILLP（230个氨基酸）的氨基酸序列与原核生物的核糖体回收因子（RRF）相似。与原核生物RRF相比，FIL IP具有N末端46氨基酸延伸，该氨基酸扩展能够充当线粒体靶向序列。 Deltafil 1菌株的亚细胞分级表明，蛋白质构成了与野生型菌株不同的deltafil 1菌株的线粒体馏分，但类似于氯霉素治疗的细胞或Rho的蛋白质。细胞。这些结果表明，FILLP对于塞维西亚链球菌线粒体中的蛋白质合成是必需的。结果表明，线粒体和细胞核之间存在通信途径，反映了编码乙氧基化循环的关键酶和线粒体呼吸链缺乏时的基因表达。较少的

项目成果

M.Ueda et al.: "Gonetic evaluation of physiological functions of thiolase isozymes in n-alkane-assinilating yeast,Candida tropicalis" J.Bacteriol.180・3. 690-698 (1998)

M.Ueda等：“正烷烃同化酵母、热带假丝酵母中硫解酶同工酶的生理功能的Gonetic评价”J.Bacteriol.180·3(1998)。

M.Ueda et al.: "Genetic evaluation of Peroxisomal and cytosolic acetoacetyl-CoA thiolase isozymes in n-alkane-assimilating diploid yeast,Citropicalis" Cell Biochem.Biophys.(in press).

M.Ueda 等人：“正烷同化二倍体酵母 Citropicalis 中过氧化物酶体和胞质乙酰乙酰辅酶 A 硫解酶同工酶的遗传评价”Cell Biochem.Biophys.（出版中）。

M.Ueda et al.: "Analysis of carbon Source-regulated gene expression by the upstream region of the Candida tropicalis malate synthase gene" Biochim.Biophys.Acta. 1350. 80-88 (1997)

M.Ueda 等人：“热带假丝酵母苹果酸合酶基因上游区域碳源调节基因表达的分析”Biochim.Biophys.Acta。

M・Ueda et al.: "A regulatory factor,Fillp,involved in derepression of isocitrate lzase gene in Saccharomyces cerevisiae:a possible mitochondrial protein" Eur.J.Biochem.256・1. 212-220 (1998)

M. Ueda 等：“调节因子 Fillp，参与酿酒酵母中异柠檬酸酶基因的去抑制：一种可能的线粒体蛋白”Eur.J.Biochem.256·1（1998）。

M.Ueda et al.: "Sorting of Peroxisomal and Mitochondrial Carnitine Acetyltransferas Isotymes in the diploid yeast,Candida tropicalis" Cell Biochem.Biophys.(in press).

M.Ueda 等人：“二倍体酵母热带假丝酵母中过氧化物酶体和线粒体肉碱乙酰转移酶同种型的分类”Cell Biochem.Biophys.（出版中）。