The PaaX Repressor, a Link between Penicillin G Acylase and the Phenylacetyl-Coenzyme A Catabolon of
            <i>Escherichia coli</i>
            W

Galán, Beatriz; Garcı́a, José Luis; Prieto, María Auxiliadora

doi:10.1128/jb.186.7.2215-2220.2004

Cited by 23 publications

(20 citation statements)

References 37 publications

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“…Direct analysis of this putative role for the PaaX regulator is the subject of further research. The work described here expands the known PaaX regulon to include the styABCD operon and supports the notion that PaaX, by sensing the PA-CoA, is a master regulatory protein in the PA-CoA catabolon that adjusts the expression of different operons to that of the paa-encoded central pathway (17). PaaX-mediated repression of the P styA promoter acts in concert with the specific StyS-StyR regulatory system to provide a dual signal-responsive system that is functionally analogous to other genetic systems involved in the degradation of toxic compounds, such as the synergist effect of a pathway substrate and intermediate in benzoate catabolism (7,8).…”

Section: Fig 3 Paax Binding Sites Comparison Of Paax Binding Sitesmentioning

confidence: 60%

“…strain Y2 contains a putative binding site for PaaX. Initially, a 15-bp consensus sequence was proposed as the binding site for PaaX at different paa promoters (14,17), but this sequence was later extended to a 39-bp region with two perfect 6-bp inverted repeats (TGATTC and GAATCA) separated by 27 nucleotides (20) as shown in Fig. 3.…”

Section: Resultsmentioning

confidence: 99%

“…3). Given that PaaX operator sequences have also been found to function within promoters of non-paa genes (17,20), we also performed an in silico analysis of the promoter region of the styrene upper catabolic genes of Pseudomonas sp. strain Y2.…”

Section: Resultsmentioning

confidence: 99%

“…Controls made with MBP alone do not retain the probe (data not shown). In E. coli, PA-CoA induces the transcription of PaaX-dependent promoters through a mechanism that involves the direct binding of PA-CoA to PaaX and consequent release of this repressor (14,17,20). Therefore, we analyzed whether a similar process might control binding of PaaX to the P styA proximal binding site.…”

Section: Fig 3 Paax Binding Sites Comparison Of Paax Binding Sitesmentioning

confidence: 99%

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Coregulation by Phenylacetyl-Coenzyme A-Responsive PaaX Integrates Control of the Upper and Lower Pathways for Catabolism of Styrene by Pseudomonas sp. Strain Y2

et al. 2006

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The P styA promoter of Pseudomonas sp. strain Y2 controls expression of the styABCD genes, which are required for the conversion of styrene to phenylacetate, which is further catabolized by the products of two paa gene clusters. Two PaaX repressor proteins (PaaX1 and PaaX2) regulate transcription of the paa gene clusters of this strain. In silico analysis of the P styA promoter region revealed a sequence located just within styA that is similar to the reported PaaX binding sites of Escherichia coli and the proposed PaaX binding sites of the paa genes of Pseudomonas species. Here we show that protein extracts from some Pseudomonas strains that have paaX genes, but not from a paaX mutant strain, can bind and retard the migration of a P styA specific probe. Purified maltose-binding protein (MBP)-PaaX1 fusion protein specifically binds the P styA promoter proximal PaaX site, and this binding is eliminated by the addition of phenylacetyl-coenzyme A. The sequence protected by MBP-PaaX1 binding was defined by DNase I footprinting. Moreover, MBP-PaaX1 represses transcription from the P styA promoter in a phenylacetyl-coenzyme A-dependent manner in vitro. Finally, the inactivation of both paaX gene copies of Pseudomonas sp. strain Y2 leads to a higher level of transcription from the P styA promoter, while heterologous expression of the PaaX1 in E. coli greatly decreases transcription from the P styA promoter. These findings reveal a control mechanism that integrates regulation of styrene catabolism by coordinating the expression of the styrene upper catabolic operon to that of the paa-encoded central pathway and support a role for PaaX as a major regulatory protein in the phenylacetyl-coenzyme A catabolon through its response to the levels of this central metabolite.

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Section: Fig 3 Paax Binding Sites Comparison Of Paax Binding Sitesmentioning

confidence: 60%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Fig 3 Paax Binding Sites Comparison Of Paax Binding Sitesmentioning

confidence: 99%

See 2 more Smart Citations

Coregulation by Phenylacetyl-Coenzyme A-Responsive PaaX Integrates Control of the Upper and Lower Pathways for Catabolism of Styrene by Pseudomonas sp. Strain Y2

et al. 2006

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“…7E). However, the core sequence of the palindrome of the MeqR2 operator site, TGACGNNCGTcA, does not resemble the consensus sequence (WWTRTGATTCGY GWT) of the operators interacting with PaaX of E. coli (49). FadR appears to recognize the palindromic consensus sequence TGGN NNNNCCA, but in this sequence the two G/C base pairs of each half site are important specificity elements (48).…”

Section: Discussionmentioning

confidence: 99%

The PaaX-Type Repressor MeqR2 of Arthrobacter sp. Strain Rue61a, Involved in the Regulation of Quinaldine Catabolism, Binds to Its Own Promoter and to Catabolic Promoters and Specifically Responds to Anthraniloyl Coenzyme A

Niewerth¹,

Parschat²,

Rauschenberg³

et al. 2012

Journal of Bacteriology

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The genes coding for quinaldine catabolism in Arthrobacter sp. strain Rue61a are clustered on the linear plasmid pAL1 in two upper pathway operons (meqABC and meqDEF) coding for quinaldine conversion to anthranilate and a lower pathway operon encoding anthranilate degradation via coenzyme A (CoA) thioester intermediates. The meqR2 gene, located immediately downstream of the catabolic genes, codes for a PaaX-type transcriptional repressor. MeqR2, purified as recombinant fusion protein, forms a dimer in solution and shows specific and cooperative binding to promoter DNA in vitro. DNA fragments recognized by MeqR2 contained a highly conserved palindromic motif, 5=-TGACGNNCGTcA-3=, which is located at positions ؊35 to ؊24 of the two promoters that control the upper pathway operons, at positions ؉4 to ؉15 of the promoter of the lower pathway genes and at positions ؉53 to ؉64 of the meqR2 promoter. Disruption of the palindrome abolished MeqR2 binding. The dissociation constants (K D ) of MeqR2-DNA complexes as deduced from electrophoretic mobility shift assays were very similar for the four promoters tested (23 nM to 28 nM). Anthraniloyl-CoA was identified as the specific effector of MeqR2, which impairs MeqR2-DNA complex formation in vitro. A binding stoichiometry of one effector molecule per MeqR2 monomer and a K D of 22 nM were determined for the effector-protein complex by isothermal titration calorimetry (ITC). Quantitative reverse transcriptase PCR analyses suggested that MeqR2 is a potent regulator of the meqDEF operon; however, additional regulatory systems have a major impact on transcriptional control of the catabolic operons and of meqR2.A rthrobacter sp. strain Rue61a, an isolate from sludge of the wastewater treatment plant of a coal tar refinery, is able to utilize quinaldine (2-methylquinoline) as a source of carbon and energy (1, 2). Methylquinolines and related N-heteroaromatic compounds are constituents of coal tar and shale oil. Quinolines are more water soluble than their homocyclic naphthalene analogs, and hence they are more readily transported to subsoil and groundwater if entering the environment, e.g., from abandoned coal processing facilities or from wood-creosoting activities. Many quinoline derivatives are considered toxic and/or mutagenic and thus are of environmental concern. However, quinaldine is used in aquaculture for anesthesia of fish (3). Interestingly, quinaldine is also a possible chemical signal in the urine of the male red fox (4) and the male ferret (5) and a component of the anal sac secretion of skunks (6). Quinoline derivatives of natural origin moreover include a plethora of alkaloids from microbial, animal, and plant sources, especially from the Rutaceae (7). A number of bacterial isolates, mainly aerobes from soil, with the ability to degrade quinoline derivatives have been described in the literature (reviewed in reference 8).Quinaldine degradation by Arthrobacter sp. Rue61a is initiated by two hydroxylation steps, catalyzed by the molybdenum enzyme quinaldine 4-oxidase...

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The penicillin G acylase production by B. megaterium is amino acid consumption dependent

Pinotti

Souza

Giordano

et al. 2006

Biotech & Bioengineering

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Aiming at to enhance the production of penicillin G acylase (PGA) by Bacillus megaterium, we have performed flasks experiments using different medium composition. Using 51 g/L of casein hydrolyzed with Alcalase and 2.7 g/L of phenylacetic acid (PhAc), the following carbon substrates were tested, individually and combined: glucose, glycerol, and lactose (present in cheese whey). Glycerol and glucose showed to be effective nutrients for the microorganism growth but delayed the PGA production. Cheese whey always increased enzyme production and cell mass. However, lactose (present in cheese whey) was not a significant carbon source for B. megaterium. PhAc, amino acids, and small peptides present in the hydrolyzed casein were the actual carbon sources for enzyme production. Replacement of hydrolyzed casein by free amino acids, 10.0 g/L, led to a significant increase in enzyme production (app. 150%), with a preferential consumption of alanine, aspartic acid, glycine, serine, arginine, threonine, lysine, and glutamic acid. A decrease of the enzyme production was observed when 20.0 g/L of amino acids were used. Using the single omission technique, it was shown that none of the 18 tested amino acids was essential for enzyme production. The use of a medium containing eight of the preferentially consumed amino acids lead to similar enzyme production level obtained when using 18 amino acids. PhAc, up to 2.7 g/L, did not inhibit enzyme production, even if added at the beginning of the cultivation.

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The PaaX Repressor, a Link between Penicillin G Acylase and the Phenylacetyl-Coenzyme A Catabolon of Escherichia coli W

Cited by 23 publications

References 37 publications

Coregulation by Phenylacetyl-Coenzyme A-Responsive PaaX Integrates Control of the Upper and Lower Pathways for Catabolism of Styrene by Pseudomonas sp. Strain Y2

Coregulation by Phenylacetyl-Coenzyme A-Responsive PaaX Integrates Control of the Upper and Lower Pathways for Catabolism of Styrene by Pseudomonas sp. Strain Y2

The PaaX-Type Repressor MeqR2 of Arthrobacter sp. Strain Rue61a, Involved in the Regulation of Quinaldine Catabolism, Binds to Its Own Promoter and to Catabolic Promoters and Specifically Responds to Anthraniloyl Coenzyme A

The penicillin G acylase production by B. megaterium is amino acid consumption dependent

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