Nucleotide sequencing and characterization of Pseudomonas putida catR: a positive regulator of the catBC operon is a member of the LysR family

Rothmel, Randi K.; Aldrich, Teri L.; Houghton, John E.; Coco, Wayne M.; Ornston, L N; Chakrabarty, Ananda M.

doi:10.1128/jb.172.2.922-931.1990

Cited by 131 publications

(94 citation statements)

References 52 publications

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“…Its deduced amino acid sequence shares high identity with LysR-type transcriptional regulators involved in benzoate or catechol catabolism such as ORF R# (55 %) of one of the two catechol gene clusters of Frateuria sp. ANA-18 (Murakami et al, 1999), catR of P. putida PRS2000 (50 %) (Houghton et al, 1995) and RB1 (48 %) (Rothmel et al, 1990), and benM (48 %) and catM (45 %) of ADP1 (Collier et al, 1998 ;RomeroArroyo et al, 1995). NK8 ORF4 was named cbeR because subsequent results showed that the gene is involved in (chloro)benzoate catabolism, and that it responds to 3CB, 4CB and benzoate, as well as cis,cismuconate.…”

Section: Cloning and Sequencing Of The Chlorobenzoate Dioxygenase Genesmentioning

confidence: 99%

The chlorobenzoate dioxygenase genes of Burkholderia sp. strain NK8 involved in the catabolism of chlorobenzoates

et al. 2001

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Burkholderia sp. NK8 grows abundantly on 3-chlorobenzoate (3CB), 4-chlorobenzoate (4CB) and benzoate. The genes encoding the oxidation of (chloro)benzoates (cbeABCD) and catechol (catA, catBC), the LysR-type regulatory gene cbeR and the gene cbeE with unknown function, all of which form a single cluster in NK8, were cloned and analysed. The protein sequence of chlorobenzoate 1,2-dioxygenase (CbeABC) is 50-65 % identical to the benzoate dioxygenase (BenABC) of Acinetobacter sp. ADP1, toluate dioxygenase (XylXYZ) of the TOL plasmid pWW0 and 2-halobenzoate dioxygenase (CbdABC) of Burkholderia cepacia 2CBS. Disruption of the cbeA gene resulted in the simultaneous loss of the ability to grow on benzoate and monochlorobenzoates, indicating the involvement of the cbeABCD genes in the degradation of these aromatics. The cbeABCD genes are preceded by catA, the gene for catechol dioxygenase. lacZ transcriptional fusion studies in Pseudomonas putida showed that catA and cbeA are co-expressed under the positive control of cbeR, a LysR-type transcriptional regulatory gene. The cbeA ::lacZ transcriptional fusion studies showed that the inducers of the genes are 3CB, 4CB, benzoate and probably cis,cis-muconate. On the other hand, 2-chlorobenzoate (2CB) did not activate the expression of the genes. The chlorobenzoate dioxygenase was able to transform 2CB, 3CB, 4CB and benzoate at considerable rates. 2CB yielded both catechol and 3-chlorocatechol (3CC), and 3CB gave rise to 4-chlorocatechol and 3CC as the major and minor intermediate products, respectively, indicating that the NK8 dioxygenase lacks absolute regiospecificity. The absence of growth of NK8 on 2CB, despite its considerable degradation activity against 2CB, is apparently due to the inability of CbeR to recognize 2CB as an inducer of the expression of the cbe genes.

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Section: Cloning and Sequencing Of The Chlorobenzoate Dioxygenase Genesmentioning

confidence: 99%

The chlorobenzoate dioxygenase genes of Burkholderia sp. strain NK8 involved in the catabolism of chlorobenzoates

et al. 2001

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“…1a) is introduced into a P. putida strain which is normally incapable of utilizing phenol, the bacteria acquire the ability to degrade phenol via the chromosomal cat pathway encoding catechol degradation in concert with the phenol to catechol or CCM conversion mediated by the pheBA operon (Kasak et al, 1993). Transcription of the chromosomal ortho pathway genes catBCA and of the plasmid-encoded pheBA operon is activated by the same positive regulator, the chromosomally encoded LysR family protein CatR (Kasak et al, 1993 ;Rothmel et al, 1990). Despite the different origins of the catBCA and the pheBA promoters, the CatR-dependent transcriptional activation mechanism of these promoters seems to be similar.…”

Section: Ipmentioning

confidence: 99%

“…Mutational analysis of the pheBA promoter (Franz & Chakrabarty, 1987) and tcbCDEF (van der Meer et al, 1991), have been characterized in bacteria capable of degrading chlorocatechols and are located on catabolic plasmids. Transcription of the catBCA, clcABD and tcbCDEF operons is controlled by the regulatory proteins CatR (Rothmel et al, 1990), ClcR (Coco et al, 1993) and TcbR (van der Meer et al, 1991), respectively. These activators belong to the LysR family of bacterial regulators and utilize similar mechanisms for activating transcription (McFall et al, 1997(McFall et al, , 1998.…”

Section: Introductionmentioning

confidence: 99%

Critical nucleotides in the interaction of CatR with the pheBA promoter: conservation of the CatR-mediated regulation mechanisms between the pheBA and catBCA operons

et al. 2000

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The promoter of the plasmid-borne pheBA genes encoding enzymes for phenol degradation resembles the catBCA promoter and is activated by CatR, the regulator of the chromosomally encoded catechol-degradative catBCA genes in Pseudomonas putida. In this study, site-directed mutagenesis of the pheBA promoter region was performed. The interrupted inverted repeat sequence of the CatR recognition binding site (RBS) of the pheBA promoter is highly homologous to that of the catBCA promoter. However, the RBS was shown not to be the sole important feature for high-affinity binding of CatR to this site. Mutagenesis of the activation binding site (ABS) of CatR, which overlaps the N35 hexamer sequence TTGGAT of the promoter, revealed that the two G nucleotides in this sequence are important for promoter activity but not for CatR binding. All other substitutions made in the ABS negatively affected both the promoter activity and CatR binding. The spacer sequence of the pheBA and catBCA promoters between the N10 and N35 hexamers is 19 bp, which is longer than optimal. However, reducing the spacer region of the pheBA promoter was not sufficient for CatR-independent promoter activation. An internal binding site (IBS) for CatR is located downstream of the transcriptional start site of the catBCA genes and it negatively regulates the operon. A similar IBS was identified in the case of the pheBA operon and tested for its functionality. The results indicate a conservation of CatR-mediated regulation mechanisms between the pheBA promoter and the catBCA promoter. This universal mechanism of CatR-mediated transcriptional activation could be of great importance in enabling catechol-degrading bacteria to expand their substrate range via horizontal transfer of the phenol degradative genes.

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“…The catBCA operon encodes three enzymes of the ortho-pathway required for benzoate catabolism, namely muconate lactonizing enzyme I, muconolactone isomerase and catechol 1,2-dioxygenase, respectively (Houghton et al, 1995). The induction of this operon, which is σ&%-independent, requires a LysR family transcriptional activator, CatR, and an inducer molecule, cis,cis-muconate (CCM), an intermediate of the ortho-pathway (Rothmel et al, 1990(Rothmel et al, , 1991. The pheB and pheA genes originating from plasmid DNA of Pseudomonas sp.…”

Section: Abbreviationsmentioning

confidence: 99%

Growth medium composition-determined regulatory mechanisms are superimposed on CatR-mediated transcription from the pheBA and catBCA promoters in Pseudomonas putida

2001

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Expression of the phenol degradation pathway in Pseudomonas putida strain PaW85 requires coordinated transcription of the plasmid-borne pheBA operon encoding catechol 1,2-dioxygenase and phenol monooxygenase, respectively, and the chromosomally encoded catechol degradation catBCA operon. Transcriptional activation from the pheBA and catBCA promoters is regulated by CatR and the catechol degradation pathway intermediate cis,cis-muconate. Here it is shown that physiological control mechanisms are superimposed on this regulatory system. Transcriptional activation from the pheBA and catBCA promoters is growth-phase-regulated in P. putida cells grown on rich medium (LB medium). CatR-mediated transcription from these promoters is silenced on rich medium until the transition from exponential to stationary phase. A slight positive effect (threefold) of stationary-phase-specific sigma factor σ S on transcription from the pheBA promoter was observed. Expression of the catBCA promoter was not influenced by the activity of this sigma factor. In contrast to rich growth medium, transcription from the pheBA and catBCA promoters in minimal medium containing a mixture of glucose and sodium benzoate was rapidly induced in exponential culture. It was shown that the presence of amino acids in the culture medium causes exponential silencing of the pheBA and catBCA promoters. The possibility that a hypothetical repressor protein could be involved in physiological control of transcription from the pheBA and catBCA promoters is discussed.

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Nucleotide sequencing and characterization of Pseudomonas putida catR: a positive regulator of the catBC operon is a member of the LysR family

Cited by 131 publications

References 52 publications

The chlorobenzoate dioxygenase genes of Burkholderia sp. strain NK8 involved in the catabolism of chlorobenzoates

The chlorobenzoate dioxygenase genes of Burkholderia sp. strain NK8 involved in the catabolism of chlorobenzoates

Critical nucleotides in the interaction of CatR with the pheBA promoter: conservation of the CatR-mediated regulation mechanisms between the pheBA and catBCA operons

Growth medium composition-determined regulatory mechanisms are superimposed on CatR-mediated transcription from the pheBA and catBCA promoters in Pseudomonas putida

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