Mutations affecting regulation of the anabolic argF and the catabolic aru genes in Pseudomonas aeruginosa PAO

Itoh, Yoshifumi; Matsui, Hideki

doi:10.1007/bf00292711

Cited by 10 publications

(27 citation statements)

References 35 publications

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“…Nutrient yeast broth (NYB) and minimal medium P (MMP) were used to cultivate P. aeruginosa as described previously (15). Luria-Bertani broth and M9 minimal medium (29) were used to grow E. coli.…”

Section: Methodsmentioning

confidence: 99%

“…Restriction digestions were carried out under the conditions recommended by the suppliers (Takara Shuzo, Kyoto, Japan; Toyobo Biochemicals, Osaka, Japan; Nippon Gene, Toyama, Japan; and New England BioLabs, Beverly, Mass. ), and other DNA techniques were described previously (15,16,29).…”

Section: Methodsmentioning

confidence: 99%

“…Previously, neither argD (ACOAT) nor aruC (SOAT) mutants had been isolated in P. aeruginosa, probably because of the existence of multiple transaminases having both ACOAT and SOAT activities. The molecular and enzymological properties of SADH, SGSD, and SGDS are largely unknown.In P. aeruginosa PAO1, at least two regulatory genes, which have been mapped at 1 and 66.5 min on the PAO1 chromosome map (13,15), modulate the induction of the aru genes and the concomitant repression of the anabolic argF and carAB genes, specifying anabolic ornithine carbamoyltransferase and carbamoylphosphate synthetase, respectively, in response to exogenous arginine (15,23). The regulatory mutations in both loci cause the inability to induce the Aru enzymes and the consequent Aru Ϫ Oru Ϫ phenotype (15,17,23).…”

mentioning

confidence: 99%

“…The regulatory mutations in both loci cause the inability to induce the Aru enzymes and the consequent Aru Ϫ Oru Ϫ phenotype (15,17,23). The regulatory…”

mentioning

confidence: 99%

“…The regulatory mutations in both loci cause the inability to induce the Aru enzymes and the consequent Aru Ϫ Oru Ϫ phenotype (15,17,23). The regulatory gene at 66.5 min appears to be within or very close to the aru cluster (13,15,17), and two types of regulatory mutations which control differently the expression of argF have been identified (15); one type (argR9906, aru-107, aru-180, and aru-292) results in constitutive expression of argF, and the other type (aru-311, aru-319, and aru-325) causes a fivefold reduction in argF expression under nonrepressive conditions but still represses argF expression in the presence of arginine. In order to study the enzymes of the AST pathway and the regulatory mechanisms underlying the inverse regulation of the arginine anabolic and catabolic genes, I have cloned and characterized the aru cluster and the adjacent argR gene.…”

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confidence: 99%

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Cloning and characterization of the aru genes encoding enzymes of the catabolic arginine succinyltransferase pathway in Pseudomonas aeruginosa

Itoh

1997

J Bacteriol

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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.The arginine succinyltransferase (AST) pathway consists of five arginine catabolic enzymes: arginine succinyltransferase (AST; encoded by aruA), N 2 -succinylarginine dihydrolase (SADH; aruB), N 2 -succinylornithine 5-aminotransferase (SOAT; aruC), N-succinylglutamate 5-semialdehyde dehydrogenase (SGSD; aruD), and N-succinylglutamate desuccinylase (SGDS; aruE) (for a review, see references 4 and 8; Fig. 1). The AST pathway was first established in Pseudomonas spp. and also occurs in Aeromonas formicans, Klebsiella aerogenes, Salmonella typhimurium, and Escherichia coli (17,27,34,35). The pathway is initiated with the succinyl-coenzyme Adependent succinylation of L-arginine and converts arginine to L-glutamate and succinate, with the concomitant liberation of ammonia and carbon dioxide (Fig. 1). In Pseudomonas aeruginosa, arginine and ornithine N 2 -succinyltransferase (AOST; aruF and aruG) catalyzes the succinylation of both L-arginine and L-ornithine (33). Ornithine succinyltransferase activity of AOST results in the formation of N 2 -succinylornithine (33, 34), which is an intermediate of the AST pathway (Fig. 1). Therefore, this pathway can serve as a common catabolic route for both arginine and ornithine utilization in P. aeruginosa (4, 8,17,34) (Fig. 1). The aru genes of P. aeruginosa are clustered in a locus origina...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

“…The regulatory mutations in both loci cause the inability to induce the Aru enzymes and the consequent Aru Ϫ Oru Ϫ phenotype (15,17,23). The regulatory…”

mentioning

confidence: 99%