Cloning and characterization of the aru genes encoding enzymes of the catabolic arginine succinyltransferase pathway in Pseudomonas aeruginosa
Cloning and characterization of the aru genes encoding enzymes of the catabolic arginine succinyltransferase pathway in Pseudomonas aeruginosa
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DOI：
10.1128/jb.179.23.7280-7290.1997
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发表时间：
1997-12-01
影响因子：
3.2
通讯作者：
Itoh, Y
中科院分区：
文献类型：
--
作者：
Itoh, Y
精氨酸琥珀酸基转移酶(AST)途径是铜绿假单胞菌在好氧条件下利用精氨酸和鸟氨酸(ARU)的主要途径。克隆了携带调节基因argR和ARU结构基因簇的铜绿假单胞菌PAO1染色体的26kb DNA片段。互补试验和核苷酸序列数据按顺序确定了argR、aruC、aruF、aruG、aruD、ARUB和Arue基因的位置。AruR、aruC、aruD、abb和arue基因分别编码ArgR调节蛋白、N-2-琥珀酰鸟氨酸5-氨基转移酶、N-琥珀酰谷氨酸5-半醛脱氢酶、N-2-琥珀精氨酸二水解酶和N-琥珀酸谷氨酸脱丁基酶，aruF和aruG基因编码精氨酸和鸟氨酸N-2-琥珀酰基转移酶的亚基(AruAI和AruAII)。此外，对位于ARU簇不同位置的转录ARU融合和极性插入突变的体内分析表明，存在三个由ArgR控制的转录单位。AruCFGDB基因似乎形成一个操纵子，从aruC上游的启动子转录而来，而Arue有自己的启动子。位于aruCFGDB操纵子上游的argR基因是编码精氨酸转运基因的另一个(AOT)操纵子的成员。在39.2~39.5min的时间内，将AST酶的氨基酸序列与位于染色体区域的AST基因所确定的同源蛋白的氨基酸序列进行比较(Kohara克隆327；GenBank/EMBL/DDJB登录号，D90818)。在这两种生物中，ARU和AST基因的总体组织结构相似，不同之处在于，大肠杆菌似乎只有一个AST基因。
The arginine succinyltransferase (AST) pathway is the major arginine and ornithine utilization (aru) pathway under aerobic conditions in Pseudomonas aeruginosa. A 26-kb DNA fragment of the P. aeruginosa PAO1 chromosome carrying the regulatory argR gene and the aru structural gene cluster was cloned. Complementation tests and nucleotide sequence data established the locations of the argR, aruC, aruF, aruG, aruD, aruB, and aruE genes, in that order. The aruR, aruC, aruD, aruB, and aruE genes specify the ArgR regulatory protein, N-2-succinylornithine 5-aminotransferase, N-succinylglutamate 5-semialdehyde dehydrogenase, N-2-succinylarginine dihydrolase, and N-succinylglutamate desuccinylase, respectively, and the aruF and aruG genes encode the subunits (AruAI and AruAII) of arginine and ornithine N-2-succinyltransferases. Furthermore, in vivo analysis of transcriptional aru fusions and of polar insertion mutations located at different sites in the aru cluster indicated the presence of three transcriptional units which are controlled by ArgR. The aruCFGDB genes appear to form an operon transcribed from a promoter upstream of aruC, whereas aruE has its own promoter. The argR gene, which is located upstream of the aruCFGDB operon, is a member of another (aot) operon coding for arginine transport genes. The deduced amino acid sequences of the AST enzymes were compared to those of homologous proteins of Escherichia coli specified by the ast genes lying in the chromosome region from 39.2 to 39.5 min (Kohara clone 327; GenBank/EMBL/DDJB accession no, D90818). The overall organization of the aru and ast genes in both organisms is similar, with the exception that E. coli appears to have a single AST gene.