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粟酒裂殖酵母编码精氨酸酶的car1基因的克隆与测序。精氨酸合成和分解代谢基因对精氨酸及相关代谢物的响应表达。

Cloning and sequencing of Schizosaccharomyces pombe car1 gene encoding arginase. Expression of the arginine anabolic and catabolic genes in response to arginine and related metabolites.

作者信息

Van Huffel C, Dubois E, Messenguy F

机构信息

Institut de Recherches du CERIA, Université Libre de Bruxelles, Belgium.

出版信息

Yeast. 1994 Jul;10(7):923-33. doi: 10.1002/yea.320100707.

Abstract

We report here the cloning and sequencing of the gene encoding arginase (car1) from Schizosaccharomyces pombe. Since no arginase-less strain exists in this organism, we cloned the gene by functional complementation of a car1 mutant strain from Saccharomyces cerevisiae. The S. pombe car1 gene encodes a 323 amino acids polypeptide sharing identity with arginases from different organisms. Measurements of arg3, arg11 and car1 mRNA under different growth conditions confirm the very weak repression by arginine of the two anabolic genes and show that the induction of arginase synthesis operates at a transcriptional level. The promoter of S. pombe car1 gene does not contain the 'arginine boxes' defined as the target of the ARGR-MCM1 proteins in the promoters of the arginine co-regulated genes in S. cerevisiae. The heterologous expression of S. pombe car1 gene in S. cerevisiae is independent of the ARGRII gene product (ArgRIIp/Arg81p). Determination of arginine, ornithine and citrulline intracellular concentrations shows the efficiency of the different controls operating in S. cerevisiae, and also indicates that in S. pombe enzyme compartmentation is not always sufficient to control the arginine metabolic flux.

摘要

我们在此报告粟酒裂殖酵母中编码精氨酸酶(car1)基因的克隆与测序。由于该生物体中不存在无精氨酸酶的菌株,我们通过对酿酒酵母的car1突变株进行功能互补来克隆该基因。粟酒裂殖酵母car1基因编码一个由323个氨基酸组成的多肽,与来自不同生物体的精氨酸酶具有同源性。在不同生长条件下对arg3、arg11和car1 mRNA的测量证实,这两个合成代谢基因受精氨酸的抑制作用非常微弱,并表明精氨酸酶合成的诱导作用在转录水平上发挥作用。粟酒裂殖酵母car1基因的启动子不包含在酿酒酵母中精氨酸共调控基因启动子中被定义为ARGR-MCM1蛋白作用靶点的“精氨酸框”。粟酒裂殖酵母car1基因在酿酒酵母中的异源表达不依赖于ARGRII基因产物(ArgRIIp/Arg81p)。对精氨酸、鸟氨酸和瓜氨酸细胞内浓度的测定显示了酿酒酵母中不同调控机制的效率,同时也表明在粟酒裂殖酵母中,酶的区室化并不总是足以控制精氨酸的代谢通量。

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