CHARACTERIZATION OF THE YEAST ARG5,6 GENE - DETERMINATION OF THE NUCLEOTIDE-SEQUENCE, ANALYSIS OF THE CONTROL REGION AND OF ARG5,6 TRANSCRIPT
CHARACTERIZATION OF THE YEAST ARG5,6 GENE - DETERMINATION OF THE NUCLEOTIDE-SEQUENCE, ANALYSIS OF THE CONTROL REGION AND OF ARG5,6 TRANSCRIPT
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DOI：
10.1007/bf00273599
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发表时间：
1991-04-01
期刊：
影响因子：
--
通讯作者：
DUBOIS, E
中科院分区：
文献类型：
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作者：
BOONCHIRD, C;MESSENGUY, F;DUBOIS, E
在酿酒酵母中，ARG5,6 基因编码乙酰谷氨酰-P 还原酶和乙酰谷氨酸激酶，这两种精氨酸合成代谢酶位于线粒体中。 两种酶的合成均由精氨酸和三种调节蛋白（ARGRI、ARGII 和 ARGRIII）协调控制。 ARG5,6 基因是通过 arg5 突变株的互补来克隆的。 对含有约 3.2 kb 的 EcoRI 片段的亚克隆进行了测序，该片段补充了精氨酸的需要。 这个 3163 bp 的序列仅包含一个 2589 个核苷酸的长开放阅读框，编码 863 个氨基酸的蛋白质。 该蛋白质的大小与 Northern 杂交确定的独特转录物的长度一致。 各种调节条件下 ARG5,6 mRNA 的测量结果显示与酶水平没有相关性。 与其他精氨酸生物合成和分解代谢基因一样，精氨酸通过三种 ARGR 蛋白的调节涉及转录后控制机制。 通过体外诱变，我们在 ARG5,6 基因的 5' 非编码区创建了点突变和缺失，这使我们能够定义 ARGR 控制的主要目标。 具体调节涉及两个区域：一个位于假定的 TATA 元件和转录起始位点之间，第二个位于该位点和第一个 ATG 之间。
In Saccharomyces cerevisiae, the ARG5,6 gene encodes acetylglutamyl-P reductase and acetylglutamate kinase, two arginine anabolic enzymes which are localized in the mitochondria. The synthesis of both enzymes is co-ordinately controlled by arginine and by three regulatory proteins (ARGRI, ARGII, and ARGRIII). The ARG5,6 gene was cloned by complementation of an arg5 mutant strain. A subclone containing an EcoRI fragment of about 3.2 kb which complements the arginine requirement was sequenced. This 3163 bp sequence contains only one long open reading frame of 2589 nucleotides encoding a protein of 863 amino acids. The size of this protein is in agreement with the length of the unique transcript determined by Northern hybridization. The measurements of ARG5,6 mRNA under various regulatory conditions show no correlation with the enzyme levels. As in other arginine biosynthetic and catabolic genes, the regulation by arginine through the three ARGR proteins thus involves a post-transcriptional control mechanism. By in vitro mutagenesis we created point mutations and deletions in the 5' non-coding region of the ARG5,6 gene which allowed us to define the primary target of ARGR control. Specific regulation involves two regions: one located between the putative TATA element and the transcriptional initiation site and the second between this site and the first ATG.