DNA sequence of the tryptophan synthase genes of Pseudomonas putida

Crawford, Irving P.; Eberly, Lee

doi:10.1016/0300-9084(89)90183-1

Cited by 22 publications

(13 citation statements)

References 34 publications

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“…The strongest evidence that trpGDC functions as a single operon is the inactivational result discussed above, but the virtual absence of intercistronic spaces, with the obligatory inclusion of the Shine-Dalgarno sequences in codons near the end of the preceding cistron, is quite characteristic of operon structure in this organism. A very similar juxtaposition of trpB and trpA is seen in the genes of the tryptophan synthase operons of P. aeruginosa (16) and P. putida (8). Similarly, codon usage favoring codons ending in G or C is seen for all the trp genes sequenced in these pseudomonads and is presumably characteristic of moderately expressed Pseudomonas genes in general.…”

Section: Discussionmentioning

confidence: 68%

DNA sequences and characterization of four early genes of the tryptophan pathway in Pseudomonas aeruginosa

et al. 1990

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Two pairs of related but easily distinguishable genes for the two subunits of anthranilate synthase have been identified in Pseudomonas aeruginosa. These were cloned, sequenced, inactivated in vitro by insertion of an antibiotic resistance cassette, and returned to the P. aeruginosa chromosome, replacing the wild-type gene. Gene replacement implicated only one of the pairs in tryptophan biosynthesis. This report describes the cloning and sequencing of the tryptophan-related gene pair, designated trpE and trpG, and presents experiments implicating their gene products in tryptophan production. DNA sequence analysis as well as growth and enzyme assays of insertionally inactivated strains indicated that trpG is the first gene in a three-gene operon that also includes trpD and trpC. Complementation of Trp auxotrophs by R-prime plasmids (T. Shinomiya, S. Shiga, and M. Kageyama, Mol. Gen. Genet., 189:382-389, 1983) We began these studies by insertionally inactivating the first 1-subunit gene cloned (7)

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Section: Discussionmentioning

confidence: 68%

DNA sequences and characterization of four early genes of the tryptophan pathway in Pseudomonas aeruginosa

et al. 1990

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“…With few exceptions, G or C was preferred in the third position for amino acids with multiple codons. As previously noted for other Pseudomonas genes (8,17,58), GGG (glycine) and CCC (proline) were avoided. Also, the use of leucine and arginine codons with C in the first position was favored.…”

Section: Resultsmentioning

confidence: 97%

“…ThepeaIJ region has a G+C content of 63%, consistent with the overall G+C content of P. putida, 62.5% (39). Codon usage is typical of Pseudomonas species (8,58). With few exceptions, G or C was preferred in the third position for amino acids with multiple codons.…”

Section: Resultsmentioning

confidence: 99%

Characterization of the genes encoding beta-ketoadipate: succinyl-coenzyme A transferase in Pseudomonas putida

Parales

Harwood

1992

J Bacteriol

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beta-Ketoadipate:succinyl-coenzyme A transferase (beta-ketoadipate:succinyl-CoA transferase) (EC 2.8.3.6) carries out the penultimate step in the conversion of benzoate and 4-hydroxybenzoate to tricarboxylic acid cycle intermediates in bacteria utilizing the beta-ketoadipate pathway. This report describes the characterization of a DNA fragment from Pseudomonas putida that encodes this enzyme. The fragment complemented mutants defective in the synthesis of the CoA transferase, and two proteins of sizes appropriate to encode the two nonidentical subunits of the enzyme were produced in Escherichia coli when the fragment was placed under the control of a phage T7 promoter. DNA sequence analysis revealed two open reading frames, designated pcaI and pcaJ, that were separated by 8 bp, suggesting that they may comprise an operon. A comparison of the deduced amino acid sequence of the P. putida CoA transferase genes with the sequences of two other bacterial CoA transferases and that of succinyl-CoA:3-ketoacid CoA transferase from pig heart suggests that the homodimeric structure of the mammalian enzyme may have resulted from a gene fusion of the bacterial alpha and beta subunit genes during evolution. Conserved functional groups important to the catalytic activity of CoA transferases were also identified.

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“…In P. aeruginosa and Pseudomonas putida, expression of the trpBA genes, encoding the two subunits of tryptophan synthase, is positively regulated by the substrate, indoleglycerol phosphate (InGP), and by the product of the trpI gene (8,10,11,30). This is in contrast to the situation in E. coli, in which the synthesis of the trpA and trpB products is not inducible but instead is directly repressed by tryptophan (51).…”

mentioning

confidence: 95%

Up-promoter mutations in the trpBA operon of Pseudomonas aeruginosa

1991

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In Pseudomonas aeruginosa, the operon encoding tryptophan synthase (trpBA) is positively regulated by the TrpI protein and an intermediate in tryptophan biosynthesis, indoleglycerol phosphate (InGP). A gene fusion in which the trpBA promoter directs expression of the Pseudomonas putida xylE gene was constructed. By using a P. putida Fl todE mutant carrying this fusion on a plasmid, three cis-acting mutations that increased xylE expression enough to allow the todE strain to grow on toluene were isolated. The level of xylE transcript from the trpBA promoter was increased in all three mutants. All three mutations are base substitutions located in the -10 region of the trpBA promoter; two of these mutations make the promoter sequence more like the Escherichia coli RNA polymerase cr70 promoter consensus sequence. The activities of the wild-type and mutant trpBA promoters, as monitored by xylE expression, were assayed in P. putida PpGl and in E. coli. The up-regulatory phenotypes of the mutants were maintained in the heterologous backgrounds, as was trpl and InGP dependence. These results indicate that the P. aeruginosa trpBA promoter has the key characteristics of a typical E. coli positively regulated promoter. The results also show that the P. aeruginosa and P. putida trpI activator gene products are functionally interchangeable.

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DNA sequence of the tryptophan synthase genes of Pseudomonas putida

Cited by 22 publications

References 34 publications

DNA sequences and characterization of four early genes of the tryptophan pathway in Pseudomonas aeruginosa

DNA sequences and characterization of four early genes of the tryptophan pathway in Pseudomonas aeruginosa

Characterization of the genes encoding beta-ketoadipate: succinyl-coenzyme A transferase in Pseudomonas putida

Up-promoter mutations in the trpBA operon of Pseudomonas aeruginosa

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