Functional Analysis and Regulation of the Divergent
            <i>spuABCDEFGH-spuI</i>
            Operons for Polyamine Uptake and Utilization in
            <i>Pseudomonas aeruginosa</i>
            PAO1

Lu, Chung‐Dar; Itoh, Yoshifumi; Nakada, Yuji; Jiang, Ying

doi:10.1128/jb.184.14.3765-3773.2002

Cited by 92 publications

(119 citation statements)

References 33 publications

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“…Besides PauB enzymes, genetic analysis from our previous studies (7,16) supported participation of a putative putrescine:pyruvate transaminase SpuC in PUT catabolism. As shown in Table 2, while PAO5008 with a lesion on spuC was partially defective in growth on PUT or CAD as the sole source of carbon or nitrogen in the liquid medium, growth on these two diamines was completely abolished in PAO5730 devoid of spuC and all four pauB genes.…”

Section: Resultsmentioning

confidence: 99%

“…Plasmids pGU101 and pGU102 were used to measure pauA2 and pauA1 promoter activities, respectively (7,16). Regulatory regions of pauA4 (PA2040) and pauA5 (PA3356) were amplified by PCR from the genomic DNA of P. aeruginosa PAO1 using the following primers: PpauA5F (5=-GGT GGA TCC GAG AAT CAA CGG CAG TAC TC-3=), PpauA5R (5=-GGT AAG CTT GAT GCA TTG CAG CAG CAC GCCA-3=), PpauA4F (5=-GGT GGA TCC TTG AAC GAT CTT GCT CTT CGT ATC-3=), and PpauA4R (5=-GGT AAG CTT GAA GGA ACA GGC TCG GCT CAG-3=).…”

Section: Methodsmentioning

confidence: 99%

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Molecular Characterization of PauR and Its Role in Control of Putrescine and Cadaverine Catabolism through the -Glutamylation Pathway in Pseudomonas aeruginosa PAO1

Chou

Peng

et al. 2013

Journal of Bacteriology

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bPseudomonas aeruginosa PAO1 grows on a variety of polyamines as the sole source of carbon and nitrogen. Catabolism of polyamines is mediated by the ␥-glutamylation pathway, which is complicated by the existence of multiple homologous enzymes with redundant specificities toward different polyamines for a more diverse metabolic capacity in this organism. Through a series of markerless gene knockout mutants and complementation tests, specific combinations of pauABCD (polyamine utilization) genes were deciphered for catabolism of different polyamines. Among six pauA genes, expression of pauA1, pauA2, pauA4, and pauA5 was found to be inducible by diamines putrescine (PUT) and cadaverine (CAD) but not by diaminopropane. Activation of these promoters was regulated by the PauR repressor, as evidenced by constitutively active promoters in the pauR mutant. The activities of these promoters were further enhanced by exogenous PUT or CAD in the mutant devoid of all six pauA genes. The recombinant PauR protein with a hexahistidine tag at its N terminus was purified, and specific bindings of PauR to the promoter regions of most pau operons were demonstrated by electromobility shift assays. Potential interactions of PUT and CAD with PauR were also suggested by chemical cross-linkage analysis with glutaraldehyde. In comparison, growth on PUT was more proficient than that on CAD, and this observed growth phenotype was reflected in a strong catabolite repression of pauA promoter activation by CAD but was completely absent as reflected by activation by PUT. In summary, this study clearly establishes the function of PauR in control of pau promoters in response to PUT and CAD for their catabolism through the ␥-glutamylation pathway.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Molecular Characterization of PauR and Its Role in Control of Putrescine and Cadaverine Catabolism through the -Glutamylation Pathway in Pseudomonas aeruginosa PAO1

Chou

Peng

et al. 2013

Journal of Bacteriology

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“…Polyamines (including putrescine and spermidine) are a group of ubiquitous polycations necessary for cell growth (Lu et al, 2002). In P. aeruginosa, putrescine can be synthesized from arginine through the arginine decarboxylase pathway (Patel et al, 2006).…”

Section: Upregulation Of N Assimilationmentioning

confidence: 99%

Analysis of the Pseudomonas putida CA-3 proteome during growth on styrene under nitrogen-limiting and non-limiting conditions

et al. 2009

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Pseudomonas putida CA-3 is a styrene-degrading bacterium capable of accumulating mediumchain-length polyhydroxyalkanoate (mclPHA) when exposed to limiting concentrations of a nitrogen source in the growth medium. Using shotgun proteomics we analysed global proteome expression in P. putida CA-3 supplied with styrene as the sole carbon and energy source under N-limiting (condition permissive for mclPHA synthesis) and non-limiting (condition non-permissive for mclPHA accumulation) growth conditions in order to provide insight into the molecular response of P. putida CA-3 to limitation of nitrogen when grown on styrene. A total of 1761 proteins were identified with high confidence and the detected proteins could be assigned to functional groups including styrene degradation, energy, nucleotide metabolism, protein synthesis, transport, stress response and motility. Proteins involved in the upper and lower styrene degradation pathway were expressed throughout the 48 h growth period under both nitrogen limitation and excess. Proteins involved in polyhydroxyalkanoate (PHA) biosynthesis, nitrogen assimilation and amino acid transport, and outer membrane proteins were upregulated under nitrogen limitation. PHA accumulation and biosynthesis were only expressed under nitrogen limitation. Nitrogen assimilation proteins were detected on average at twofold higher amounts under nitrogen limitation. Expression of the branched-chain amino acid ABC transporter was up to 16-fold higher under nitrogen-limiting conditions. Branched chain amino acid uptake by nitrogenlimited cultures was also higher than that by non-limited cultures. Outer membrane lipoproteins were expressed at twofold higher levels under nitrogen limitation. This was confirmed by Western blotting (immunochemical detection) of cells grown under nitrogen limitation. Our study provides the first global description of protein expression changes during growth of any organism on styrene and accumulating mclPHA (nitrogen-limited growth). INTRODUCTIONPseudomonas putida CA-3 has been reported to utilize styrene as a sole source of carbon and energy and to accumulate the biodegradable polymer medium-chainlength polyhydroxyalkanoate (mclPHA) when nitrogen (N) becomes limited in the growth medium (O'Connor et al., 1996;. These two abilities have been employed in the two-step chemo-biotechnological conversion of polystyrene, a non-biodegradable polymer, to mclPHA at laboratory scale (Ward et al., 2006). The process has been improved through the controlled feeding of N to the growth medium, which results in a twofold increase in biomass and a 1.4-fold increase in mclPHA accumulation (Goff et al., 2007).Molecular investigations of the styrene degradation pathway in this organism have been reported (Mooney et al., 2006b;O'Connor et al., 2001). Styrene metabolism in P. putida CA-3 proceeds via initial side chain oxidation and involves an upper pathway converting styrene to phenylacetic acid (PA) (O'Connor et al., 1995), and a lower pathway initiated via activation of PA to phenylace...

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“…For the ADC pathway, although the presence of arginine-inducible ADC activity in the conversion of arginine into agmatine has been reported in an early study (29), the corresponding gene has not been identified. Recent studies in this laboratory have reported the characterization of the aguBA and spuC genes, which encode enzymes for the conversion of agmatine into 4-aminobutyrate of the ADC pathway (26,34). Exogenous agmatine but not arginine induced these genes (14,29).…”

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confidence: 99%

Transcriptome Analysis of the ArgR Regulon inPseudomonas aeruginosa

Yang

2004

J Bacteriol

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Arginine metabolism in pseudomonads with multiple catabolic pathways for its utilization as carbon and nitrogen sources is of particular interest as the model system to study control of metabolic integration. We performed transcriptome analyses to identify genes controlled by the arginine regulatory protein ArgR and to better understand arginine metabolic pathways of P. aeruginosa. We compared gene expression in wild-type strain PAO1 with that in argR mutant strain PAO501 grown in glutamate minimal medium in the presence and absence of arginine. Ten putative transcriptional units of 28 genes were inducible by ArgR and arginine, including all known ArgR-regulated operons under aerobic conditions. The newly identified genes include the putative adcAB operon, which encodes a catabolic arginine decarboxylase and an antiporter protein, and PA0328, which encodes a hypothetical fusion protein of a peptidase and a type IV autotransporter. Also identified as members of the arginine network are the following solute transport systems: PA1971 (braZ) for branched-chain amino acids permease; PA2042 for a putative sodium:serine symporter; PA3934, which belongs to the family of small oligopeptide transporters; and PA5152-5155, which encodes components of an ABC transporter for a putative opine uptake system. The effect of arginine on the expression of these genes was confirmed by lacZ fusion studies and by DNA binding studies with purified ArgR. Only five transcriptional units of nine genes were qualified as repressible by ArgR and arginine, with three operons (argF, carAB, and argG) in arginine biosynthesis and two operons (gltBD and gdhA) in glutamate biosynthesis. These results indicate that ArgR is important in control of arginine and glutamate metabolism and that arginine and ArgR may have a redundant effect in inducing the uptake systems of certain compounds.Pseudomonas aeruginosa possesses four different catabolic pathways for utilization of arginine (11): the arginine deiminase (ADI) pathway, the arginine succinyltransferase (AST) pathway, the arginine decarboxylase (ADC) pathway, and the arginine dehydrogenase (ADH) pathway (Fig. 1). Under aerobic conditions, Haas and coworkers have established that arginine utilization occurs mainly through the AST pathway, which converts arginine to glutamate (20,43). Recent studies in the laboratories of Lu and Abdelal have shown that the aru operon, which encodes the AST pathway, and the gdhB gene, which encodes a catabolic glutamate dehydrogenase, are inducible by arginine and that this effect is mediated by ArgR (18,24). In P. aeruginosa, ArgR, the arginine-responsive regulator protein, is autoinduced from the aot-argR operon for arginine uptake and regulation (35). The ArgR protein of P. aeruginosa belongs to the AraC/XylS family of transcriptional regulators (7) and is thus quite different in structure and function from the ArgR proteins of enteric bacteria and Bacillus subtilis (3,4,6,23,25), which have a high degree of similarity in their three-dimensional structures and DNA...

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Functional Analysis and Regulation of the Divergent spuABCDEFGH-spuI Operons for Polyamine Uptake and Utilization in Pseudomonas aeruginosa PAO1

Cited by 92 publications

References 33 publications

Molecular Characterization of PauR and Its Role in Control of Putrescine and Cadaverine Catabolism through the -Glutamylation Pathway in Pseudomonas aeruginosa PAO1

Molecular Characterization of PauR and Its Role in Control of Putrescine and Cadaverine Catabolism through the -Glutamylation Pathway in Pseudomonas aeruginosa PAO1

Analysis of the Pseudomonas putida CA-3 proteome during growth on styrene under nitrogen-limiting and non-limiting conditions

Transcriptome Analysis of the ArgR Regulon inPseudomonas aeruginosa

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