The Target for the
            <i>Pseudomonas putida</i>
            Crc Global Regulator in the Benzoate Degradation Pathway Is the BenR Transcriptional Regulator

Moreno, Renata; Rojo, Fernando

doi:10.1128/jb.01604-07

Cited by 90 publications

(112 citation statements)

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“…Second, Crc inhibits the translation of some key structural genes of the pathway. Most notably, Crc targets were found not only at the 5Ј-end of the regulated mRNAs, as observed for other Crc-regulated genes (31,32,33), but also at internal positions in the upper and meta polycistronic mRNAs. Fig.…”

Section: Discussionmentioning

confidence: 72%

“…The benzoate generated from toluene can be degraded through the pWW0-encoded meta pathway genes or through the chromosomally encoded benzoate ortho pathway because this "chromosomal" pathway is also induced in the presence of upper pathway substrates (15,47). However, Crc also exerts a strong repressing effect on expression of the chromosomal ortho pathway for benzoate (6,31).…”

Section: Discussionmentioning

confidence: 99%

“…4). Purified Crc can bind in vitro short RNA fragments containing a proper target; the specificity of this binding has been demonstrated by competition and mutagenesis assays (30,31,32). Band-shift assays were therefore used to analyze the ability of purified Crc to bind radioactively labeled run-off RNA fragments corresponding to the 5Ј-end of the mRNAs originating at promoters Pu, Pm, P S1 , and P R1 .…”

Section: Resultsmentioning

confidence: 99%

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The Crc Global Regulator Inhibits the Pseudomonas putida pWW0 Toluene/Xylene Assimilation Pathway by Repressing the Translation of Regulatory and Structural Genes

Moreno

Fonseca²,

Rojo

2010

Journal of Biological Chemistry

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In Pseudomonas putida, the expression of the pWW0 plasmid genes for the toluene/xylene assimilation pathway (the TOL pathway) is subject to complex regulation in response to environmental and physiological signals. This includes strong inhibition via catabolite repression, elicited by the carbon sources that the cells prefer to hydrocarbons. The Crc protein, a global regulator that controls carbon flow in pseudomonads, has an important role in this inhibition. Crc is a translational repressor that regulates the TOL genes, but how it does this has remained unknown. This study reports that Crc binds to sites located at the translation initiation regions of the mRNAs coding for XylR and XylS, two specific transcription activators of the TOL genes. Unexpectedly, eight additional Crc binding sites were found overlapping the translation initiation sites of genes coding for several enzymes of the pathway, all encoded within two polycistronic mRNAs. Evidence is provided supporting the idea that these sites are functional. This implies that Crc can differentially modulate the expression of particular genes within polycistronic mRNAs. It is proposed that Crc controls TOL genes in two ways. First, Crc inhibits the translation of the XylR and XylS regulators, thereby reducing the transcription of all TOL pathway genes. Second, Crc inhibits the translation of specific structural genes of the pathway, acting mainly on proteins involved in the first steps of toluene assimilation. This ensures a rapid inhibitory response that reduces the expression of the toluene/ xylene degradation proteins when preferred carbon sources become available.

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Section: Discussionmentioning

confidence: 72%

Section: Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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The Crc Global Regulator Inhibits the Pseudomonas putida pWW0 Toluene/Xylene Assimilation Pathway by Repressing the Translation of Regulatory and Structural Genes

Moreno

Fonseca²,

Rojo

2010

Journal of Biological Chemistry

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“…The levels and activity of Crc vary depending on growth conditions [231] and it has also been reported to inhibit the expression of many other catabolic pathways for several non-preferred compounds in Pseudomonads [230,[232][233][234][235][236][237].…”

Section: Catabolite Repression Controlmentioning

confidence: 99%

Monooxygenases involved in the n-alkanes metabolism by Rhodococcus sp. BCP1: molecular characterization and expression of alkB gene

Cappelletti

Fedi

Honda

et al. 2010

Journal of Biotechnology

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Discussion 107 Summary 113Chapter 5 -Analysis of the alkB gene expression Summary 157Chapter 6 -Proteomic analysis of n-alkanes growth of Rhodococcus sp. BCP1 Chapter 1 -General Introduction PrefaceThe quality of life on Earth is tightly related with the overall quality of the environment [1]. During the last century it was commonly believed that the abundance of land and resources were unlimited and it was rarely acknowledged that the production, use, and disposal of hazardous substances had environmental and health effects [1][2][3]. Owing to this, human activity has deliberately or inadvertently released into waters and soil a variety of toxic compounds as refrigerants, paints, solvents, herbicides and pesticides that can cause considerable environmental pollution and human health problems as a result of their persistence, toxicity, and transformation into hazardous metabolites [4,5]. Very soon, however, it became clear how the carelessness and negligence in handling the industrial wastes and gas emissions had caused dramatic effect on the nature and on human health [1].Nowadays, the concern about the global warming and the climate changing induced the government regulatory agencies to establish procedures for assessing the environmental impact and health hazards of chemicals and for controlling/limiting their utilization.International efforts have been applied to remedy many contaminated sites all over the world, either as a response to the risk of adverse health or environmental effects caused by contamination or to enable the site to be redeveloped for use [5]. In response to public and government concern and because of the intriguing research problems presented, environmental scientists, biologists and chemists have been giving increased attention to identifying and determining the behavior and fate of organic compounds in natural ecosystems [5].Bioremediation procedures offer an economical and effective possibility to degrade or render innocuous toxic pollutants using natural biological activity [1]. By definition, bioremediation implies the use of living organisms, primarily microorganisms, to degrade the Since contaminant compounds are transformed by living organisms through reactions that take place as a part of their metabolic processes, environmental biotechnology focuses on the development of techniques to optimize environmental parameters to allow these metabolic pathways to proceed faster and, therefore, to improve the bio-degradation [1]. As a result, the removal of the contaminated soil can be necessary to let the biodegradation proceed in controlled sites (such as bioreactors). These ex situ approaches provide optimal conditions out one single step of the entire bio-degradation process. Hydrocarbons in the environmentHydrocarbons are energy-rich organic compounds consisting of carbon and hydrogen atoms and they can be saturated (only single C-H bounds) or unsaturated (one ore more double or triple C-H bounds). Alkanes are saturated hydrocarbons with the general formula C n H 2n+2 ; they can b...

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“…This carbon catabolite repression (CCR) mechanism of PAA metabolism in P. putida is not yet fully understood. An RNA-binding global regulator in P. putida, Crc (Moreno & Rojo, 2008), appears to be involved in CCR of the degradation pathways for several aromatic compounds, such as benzoate and 4-hydroxybenzoate, but not PAA (Morales et al, 2004). In E. coli, the cAMP receptor protein Crp is involved in CCR of PAA degradation (Ferrández et al, 2000;Kimata et al, 1997;Notley-McRobb et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Intracellular 2-keto-3-deoxy-6-phosphogluconate is the signal for carbon catabolite repression of phenylacetic acid metabolism in Pseudomonas putida KT2440

et al. 2009

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The growth pattern of Pseudomonas putida KT2440 in the presence of glucose and phenylacetic acid (PAA), where the sugar is used in preference to the aromatic compound, suggests that there is carbon catabolite repression (CCR) of PAA metabolism by glucose or gluconate. Furthermore, CCR is regulated at the transcriptional level. However, this CCR phenomenon does not occur in PAA-amended minimal medium containing fructose, pyruvate or succinate. We previously identified 2-keto-3-deoxy-6-phosphogluconate (KDPG) as an inducer of glucose metabolism, and this has led to this investigation into the role of KDPG as a signal compound for CCR. Two mutant strains, the edd mutant (non-KDPG producer) and the eda mutant (KDPG overproducer), grew in the presence of PAA but not in the presence of glucose. The edd mutant utilized PAA even in the presence of glucose, indicating that CCR had been abolished. This observation has additional support from the finding that there is high phenylacetyl-CoA ligase activity in the edd mutant, even in the presence of glucose+PAA, but not in wild-type cells under the same conditions. Unlike the edd mutant, the eda mutant did not grow in the presence of glucose+PAA. Interestingly, there was no uptake and/or metabolism of PAA in the eda mutant cells under the same conditions. Targeted disruption of PaaX, a repressor of the PAA operon, had no effect on CCR of PAA metabolism in the presence of glucose, suggesting that there is another transcriptional repression system associated with the KDPG signal. This is the first study to demonstrate that KDPG is the true CCR signal of PAA metabolism in P. putida KT2440. INTRODUCTIONIn Pseudomonas putida, the phenylacetic acid (PAA) catabolic gene clusters consist of 15 genes that are organized into five contiguous operons (del Peso-Santos et al., 2008; Di Gennaro et al., 2007; Jiménez et al., 2002;Luengo et al., 2001;Olivera et al., 1998). The PAA pathway ( Fig. 1) is a major route of a complex functional unit that catalyses the transformation of structurally related compounds such as styrene, tropic acid, ethylbenzene and 2-phenylethylalanine into phenylacetyl-CoA (PA-CoA), a common intermediate (Luengo et al., 2001). It has been reported that PA-CoA, and not PAA, is the true inducer of the PAA catabolic pathway and that the PaaX repressor controls PAA-catabolic gene expression (Ferrández et al., 2000;García et al., 2000). Binding of PaaX to its cognate DNA binding sites is abolished when the inducer, PA-CoA, interacts with the PaaX protein (Bartolomé-Martín et al., 2004; del Peso-Santos et al., 2006; Ferrández et al., 2000;García et al., 2000). Interestingly, along with the StyR/StyS system, PaaX also regulates the styrene degradation pathway by binding to the promoter region of the styrene upper pathway in Pseudomonas sp. strain Y2 (del PesoSantos et al., 2006). PaaX is also involved in the transcriptional regulation of the pac gene, which encodes penicillin G acylase in Escherichia coli W (Galán et al., 2004;Kim et al., 2004). PaaX is involved in the co...

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The Target for the Pseudomonas putida Crc Global Regulator in the Benzoate Degradation Pathway Is the BenR Transcriptional Regulator

Cited by 90 publications

References 46 publications

The Crc Global Regulator Inhibits the Pseudomonas putida pWW0 Toluene/Xylene Assimilation Pathway by Repressing the Translation of Regulatory and Structural Genes

The Crc Global Regulator Inhibits the Pseudomonas putida pWW0 Toluene/Xylene Assimilation Pathway by Repressing the Translation of Regulatory and Structural Genes

Monooxygenases involved in the n-alkanes metabolism by Rhodococcus sp. BCP1: molecular characterization and expression of alkB gene

Intracellular 2-keto-3-deoxy-6-phosphogluconate is the signal for carbon catabolite repression of phenylacetic acid metabolism in Pseudomonas putida KT2440

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