Arginine Analogues: Effect on Growth and on the First Two Enzymes of the Arginine Pathway in Pseudomonas aeruginosa

Leisinger, Thomas; O'sullivan, Christine Y.; Haas, D

doi:10.1099/00221287-84-2-253

Cited by 17 publications

(8 citation statements)

References 4 publications

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“…While inactivation of a gene required for arginine transport could result in insensitivity to exogenous arginine, this is not the case here, since P. aeruginosa, like P. putida (28), possesses a general basic amino acid transport system in addition to the specific arginine transport system. Rather, gel retardation experiments with various subclones localized, on a 1.3-kb fragment downstream of aotJ, a gene encoding a DNA-binding protein, that interacts with the control regions for the car and argF operons.…”

Section: Discussionmentioning

confidence: 81%

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Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1

Park

Abdelal

1997

J Bacteriol

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Gel retardation experiments indicated the presence in Pseudomonas aeruginosa cell extracts of an arginineinducible DNA-binding protein that interacts with the control regions for the car and argF operons, encoding carbamoylphosphate synthetase and anabolic ornithine carbamoyltransferase, respectively. Both enzymes are required for arginine biosynthesis. The use of a combination of transposon mutagenesis and arginine hydroxamate selection led to the isolation of a regulatory mutant that was impaired in the formation of the DNA-binding protein and in which the expression of an argF::lacZ fusion was not controlled by arginine. Experiments with various subclones led to the conclusion that the insertion affected the expression of an arginine regulatory gene, argR, that encodes a polypeptide with significant homology to the AraC/XylS family of regulatory proteins. Determination of the nucleotide sequence of the flanking regions showed that argR is the sixth and terminal gene of an operon for transport of arginine. The argR gene was inactivated by gene replacement, using a gentamicin cassette. Inactivation of argR abolished arginine control of the biosynthetic enzymes encoded by the car and argF operons. Furthermore, argR inactivation abolished the induction of several enzymes of the arginine succinyltransferase pathway, which is considered the major route for arginine catabolism under aerobic conditions. Consistent with this finding and unlike the parent strain, the argR::Gm derivative was unable to utilize arginine or ornithine as the sole carbon source. The combined data indicate a major role for ArgR in the control of arginine biosynthesis and aerobic catabolism.Arginine metabolism is of considerable significance in Pseudomonas aeruginosa, which can utilize this amino acid as a good source of carbon, nitrogen, and energy (16). The significance of arginine as a nutrient to P. aeruginosa is reflected in its being one of the strongest chemotactic attractants for this organism (10). This significance is also reflected in the presence of four catabolic pathways for the utilization of arginine ( Fig. 1) (16): the arginine deiminase pathway, the arginine succinyltransferase (AST) pathway; the arginine dehydrogenase pathway, and the arginine decarboxylase pathway.Recent work by Haas and coworkers has elucidated the role of anr (for anaerobic regulation) in induction of the arginine deiminase pathway by low oxygen tension (15). The arginine deiminase pathway functions to provide P. aeruginosa with energy in the absence of appropriate terminal electron acceptors (43). The anr gene of P. aeruginosa appears to be a close relative of fnr of Escherichia coli (49); in fact, the two genes can replace each other in heterologous systems (15). In contrast to the current understanding of the molecular basis for control of the arginine deiminase pathway, analogous information is lacking regarding the AST pathway (Fig. 1). This pathway, which converts arginine to glutamate, is considered the major route for catabolism in P. aeruginosa u...

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Section: Discussionmentioning

confidence: 81%

“…Arginine hydroxamate was successfully used by Mountain and Baumberg (33) for selection of arginine regulatory mutants of Bacillus subtilis. Since this arginine analog was reported to also inhibit the growth of P. aeruginosa (28), it appeared promising for use in the selection of mutants defective in arginine regulation in this organism.…”

Section: Resultsmentioning

confidence: 99%

Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1

Park

Abdelal

1997

J Bacteriol

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“…1). Inhibition of the first two enzymes of arginine biosynthesis by arginine, by arginine analogues and by several other effectors has been observed with purified enzyme preparations from P. aeruginosa Leisinger et al, 1974;Haas & Leisinger, 1975). In the same organism Isaac &rHolloway (1972) found up to 100-fold repression of the anabolic ornithine carbamoyltransferase ('enzyme 6, 1971).…”

Section: Introductionmentioning

confidence: 99%

Regulation of Enzyme Synthesis in the Arginine Biosynthetic Pathway of Pseudomonas aeruginosa

Voellmy¹,

Leisinger²

1978

Journal of General Microbiology

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In Pseudomonas aeruginosa the synthesis of only two out of eight arginine biosynthetic enzymes tested was regulated. Comparisons were made between the specific activities of these enzymes in bacteria grown on arginine or on its precursor, glutamate. N2-Acetylornithine 5-aminotransferase (ACOAT), an enzyme involved in both the biosynthesis and catabolism of arginine, was induced about 14-fold during growth of the organism on arginine as the only carbon and nitrogen source, and the anabolic ornithine carbamoyltransferase (aOTC), a strictly biosynthetic enzyme, was repressed 18-fold. Addition of various carbon sources to the arginine medium led to repression of ACOAT and to derepression of aOTC. Fructose, which supported only slow growth of P. arruginosa, had a weak regulatory effect on the synthesis of the two arginine enzymes while citrate, a good carbon source for this organism, had a strong effect. The repression of ACOAT by citrate was not relieved by adding cyclic AMP to the medium. Under a variety of growth conditions leading to different enzyme activities, a linear relationship between the reciprocal of the specific activity of ACOAT and the specific activity of aOTC was observed. This inverse regulation of the formation of the two enzymes suggested that a single regulatory system governs their synthesis. Such a view was supported by the isolation of citrate-resistant regulatory mutants which constitutively formed ACOAT at the induced level and aOTC at the repressed level.

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“…The sensitivity of P. aeruginosa to arginine analogs (Lindospicine and L-arginine hydroxamate) was J. BACTERIOL. also found to vary with the source of carbon supplied to support growth (7). In Neurospora crassa, a comparison of enzymological results obtained after growth on various carbon sources provided insight into the regulation of glycerol kinase (5).…”

Section: Resultsmentioning

confidence: 97%

Increased antimetabolite sensitivity with variation of carbon source during growth

Jensen¹,

Calhoun²

1978

J Bacteriol

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In Serratia marcescens, analogs of leucine (norleucine), methionine (a-methylmethionine), histidine (3-amino-1,2,4-triazolealanine), tyrosine (p-aminophenylalanine), and tryptophan (7-methylindole) are conditional inhibitors of growth; inhibition occurs during the metabolism of some carbon sources but not with others. A further increase in sensitivity to growth inhibition by these analogs can be accomplished through the use of particular combinations of carbon sources present in the inoculum and in the subsequent analog-containing culture medium. Variable sensitivity to analog-mediated inhibition of growth observed during growth on glucose, glycerol, fructose, or citrate correlated inversely with the intracellular pool sizes of the amino acids cognate to the analogs used. The abovecited results, in conjunction with previous results obtained with Pseudomonas aeruginosa and Bacillus subtilis, involve diverse biochemical pathways and suggest that nutritional manipulation to alter the pattern of carbon flow in microorganisms is a generally useful means to accomplish increased sensitivity to growth inhibition by metabolite analogs.

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Arginine Analogues: Effect on Growth and on the First Two Enzymes of the Arginine Pathway in Pseudomonas aeruginosa

Cited by 17 publications

References 4 publications

Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1

Cloning and characterization of argR, a gene that participates in regulation of arginine biosynthesis and catabolism in Pseudomonas aeruginosa PAO1

Regulation of Enzyme Synthesis in the Arginine Biosynthetic Pathway of Pseudomonas aeruginosa

Increased antimetabolite sensitivity with variation of carbon source during growth

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