Control of pyrimidine formation in Pseudomonas putida ATCC 17536

2002 ◽  
Vol 48 (12) ◽  
pp. 1076-1081 ◽  
Author(s):  
Manuel F Santiago ◽  
Thomas P West
Keyword(s):  
Carbon Source ◽  
Pseudomonas Putida ◽  
Orotic Acid ◽  
De Novo ◽  
Enzyme Synthesis ◽  
Pyrimidine Biosynthesis ◽  
Transcriptional Level ◽  
Orotate Phosphoribosyltransferase ◽  
Biosynthesis Regulation ◽  
Uracil Auxotrophs

The regulation of de novo pyrimidine biosynthesis in Pseudomonas putida ATCC 17536 by pyrimidines was explored. The pathway enzyme activities were higher in glucose-grown cells than in succinate-grown cells, indicating catabolite repression by succinate. In P. putida cells grown on succinate as a carbon source, only aspartate transcarbamoylase activity was greatly diminished by uracil supplementation. When glucose was the carbon source, orotic acid supplementation significantly decreased orotate phosphoribosyltransferase and orotidine 5'-monophosphate (OMP) decarboxylase activities. Uracil auxotrophs, deficient for dihydroorotase activity or with reduced phosphoribosyltransferase activity, were isolated. After pyrimidine limitation of both auxotrophs, the greatest derepression of enzyme activity was observed for OMP decarboxylase independent of carbon source. Orotic acid induced both phosphoribosyltransferase and decarboxylase activities in glucose-grown cells of the dihydroorotase-deficient strain. Regulation at the transcriptional level of de novo pyrimidine biosynthetic enzyme synthesis in P. putida ATCC 17536 was observed, which contrasts with previous observations.Key words: pyrimidine biosynthesis, regulation, auxotrophs, induction, Pseudomonas.

Regulation of pyrimidine formation in Pseudomonas lundensis

2009 ◽  
Vol 55 (3) ◽  
pp. 261-268 ◽  
Author(s):  
Thomas P. West
Keyword(s):  
Carbon Source ◽  
Orotic Acid ◽  
De Novo ◽  
Enzyme Synthesis ◽  
Chemical Mutagenesis ◽  
Aspartate Transcarbamoylase ◽  
Food Spoilage ◽  
Orotate Phosphoribosyltransferase ◽  
Selection Procedures ◽  
Monophosphate Decarboxylase

The regulation of pyrimidine formation in the food spoilage agent Pseudomonas lundensis ATCC 49968 by pyrimidines was examined. In P. lundensis cells grown on glucose as a carbon source, the enzymes aspartate transcarbamoylase, dihydroorotase, and orotidine 5′-monophosphate decarboxylase were induced by orotic acid. Pyrimidine auxotrophs containing reduced transcarbamoylase or orotate phosphoribosyltransferase activity were isolated using chemical mutagenesis and selection procedures. Independent of carbon source, the maximum derepression of enzyme activity was observed for orotidine 5′-monophosphate decarboxylase after pyrimidine limitation of either auxotroph. In the glucose-grown cells of the transcarbamoylase mutant strain, orotic acid induced dihydroorotase and decarboxylase activities. Aspartate transcarbamoylase activity in succinate-grown P. lundensis cells was highly regulated by pyrophosphate as well as by pyrimidine and purine ribonucleotides. It was concluded that pyrimidine formation in P. lundensis was controlled both at the level of de novo pyrimidine biosynthetic enzyme synthesis and at the level of transcarbamoylase activity.

Induction and repression of L-fucose dehydrogenase of Pullularia pullulans

1984 ◽  
Vol 30 (6) ◽  
pp. 753-757 ◽  
Author(s):  
P. F. Conter ◽  
M. F. Guimarães ◽  
L. A. Veiga
Keyword(s):  
Carbon Source ◽  
Sole Carbon Source ◽  
De Novo ◽  
Ring Structure ◽  
Enzyme Synthesis ◽  
De Novo Synthesis

The growth of Pullularia pullulans on L-fucose as the sole carbon source induces the synthesis of L-fucose dehydrogenase, a NAD-dependent enzyme that catalyzes the oxidation of L-fucose to L-fucono-δ-lactone, which spontaneously hydrolyzes to L-fuconic acid. The induction of the enzyme is inhibited by cycloheximide, suggesting de novo synthesis. D-Glucose, D-galactose, and glycerol at 0.5% concentration gave rise to 62, 54, 51, and 46% of repression of enzyme synthesis, respectively. No repression effect was detected with D-arabinose and L-rhamnose. L-Arabinose repressed only 20% of the enzyme synthesis. Among the sugars tested, only L-fucose and L-galactose were oxidized by the enzyme. L-Galactose, which shares a common ring structure with L-fucose, showed only 10% of the activity observed when L-fucose was used.

Fetal fuels. V. Ketone bodies inhibit pyrimidine biosynthesis in fetal rat brain

1982 ◽  
Vol 243 (3) ◽  
pp. E234-E239
Author(s):  
S. Bhasin ◽  
G. E. Shambaugh
Keyword(s):  
Rat Brain ◽  
Orotic Acid ◽  
Ketone Body ◽  
De Novo ◽  
Ketone Bodies ◽  
Pyrimidine Biosynthesis ◽  
De Novo Biosynthesis ◽  
Fetal Rat ◽  
Beta Hydroxybutyrate ◽  
Fetal Rat Brain

Ketonemic states complicating late pregnancy are accompanied by lower brain weights in the newborn. Potential mechanisms whereby ketone bodies might inhibit cell proliferation were therefore examined in the fetal rat brain slice by measuring their impact on the de novo pathway for pyrimidine biosynthesis. DL-beta-hydroxybutyrate (10.8 mM) and acetoacetate (5.4 mM) were both found to diminish the incorporation of NaH14CO3 into [14C]UMP by 30%. This effect was similar in fetal tissues from fed and 48-h starved mothers. Graded concentrations of DL-beta-hydroxybutyrate (1.4-43.2 mM) resulted in a progressive inhibition that could not be explained either by isotope dilution consequent to ketone body oxidation or by a generalized inhibition of protein synthesis. The inhibition was not reversed with 10 mM glutamine, the principal nitrogen substrate for de novo biosynthesis of pyrimidines. When the conversion of orotic acid into UMP was blocked with 6-azauridine, DL-beta-hydroxybutyrate (10.8 mM) inhibited the incorporation of NaH14CO3 into orotic acid by 28%. By contrast, maximally inhibitory concentrations of this ketone body (43.2 mM) had no effect on the incorporation of [6-14C]orotic acid into [14C]UMP. Is is concluded that ketone bodies inhibit the de novo biosynthesis of pyrimidines in fetal brain slices and that they do so at a site proximal to orotic acid formation.

Pyrimidine nucleotide synthesis inPseudomonascitronellolis

2004 ◽  
Vol 50 (6) ◽  
pp. 455-459 ◽  
Author(s):  
Thomas P West
Keyword(s):  
Enzyme Activities ◽  
De Novo ◽  
Pyrimidine Biosynthesis ◽  
Aspartate Transcarbamoylase ◽  
Decarboxylase Activity ◽  
Wild Type ◽  
Nucleotide Synthesis ◽  
Pyrimidine Nucleotide ◽  
Mutant Cells ◽  
Biosynthesis Regulation

Pyrimidine biosynthesis was active in Pseudomonas citronellolis ATCC 13674 and appeared to be regulated by pyrimidines. When wild-type cells were grown on succinate in the presence of uracil, the de novo enzyme activities were depressed while only four enzyme activities were depressed in the glucose-grown cells. On either carbon source, orotic acid-grown cells had diminished aspartate transcarbamoylase, dihydroorotase or OMP decarboxylase activity. Pyrimidine limitation of glucose-grown pyrimidine auxotrophic cells resulted in de novo enzyme activities, except for transcarbamoyolase activity, that were elevated by more than 5-fold compared to their activities in uracil-grown cells. Since pyrimidine limitation of succinate-grown mutant cells produced less enzyme derepression, catabolite repression appeared to be a factor. At the level of enzyme activity, aspartate transcarbamoylase activity in P. citronellolis was strongly inhibited by all effectors tested. Compared to the regulation of pyrimidine biosynthesis in taxonomically-related species, pyrimidine biosynthesis in P. citronellolis appeared more highly regulated.Key words: pyrimidine biosynthesis, regulation, Pseudomonas citronellolis, auxotroph, aspartate transcarbamoylase, inhibition.

De novo production of geranic acid with Pseudomonas putida

2014 ◽  
Vol 31 ◽  
pp. S72-S73
Author(s):  
Jens Schrader ◽  
Jia Mi ◽  
Daniela Becher ◽  
Patrice Lubuta ◽  
Markus Buchhaupt ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
De Novo ◽  
Geranic Acid

Transcriptional regulation of the human CAD gene during myeloid differentiation

1987 ◽  
Vol 7 (5) ◽  
pp. 1961-1966
Author(s):  
G N Rao ◽  
E S Buford ◽  
J N Davidson
Keyword(s):  
Gene Expression ◽  
De Novo ◽  
Morphological Changes ◽  
Transcriptional Level ◽  
Carbamoyl Phosphate ◽  
Aspartate Transcarbamylase ◽  
Trans Retinoic Acid ◽  
Beta Actin ◽  
Cad Gene ◽  
Cad Protein

CAD codes for a trifunctional protein involved in the catalysis of the first three enzymatic activities in the de novo pyrimidine biosynthetic pathway, namely, carbamoyl-phosphate synthetase II (EC 6.3.5.5), aspartate transcarbamylase (EC 2.1.3.2), and dihydroorotase (EC 3.5.2.3). CAD regulation was studied in the human promyelocyte leukemic line HL-60 as it differentiated into monocytic or granulocytic lineages after induction by 12-O-tetradecanoylphorbol-13-acetate or trans-retinoic acid and dibutyryl cyclic AMP, respectively. Within 12 h of induction of HL-60 cells with either inducer, total cellular levels of CAD RNA essentially disappeared. On the other hand, no apparent decreases in beta-actin RNA levels were seen even 48 h after HL-60 cells were induced, as compared with untreated cells. With nuclear runoff assays, it was clearly shown that the inactivation of CAD gene expression during the induction of HL-60 cells with either inducer was at the transcriptional level. The nuclear runoff experiments also demonstrated that the CAD gene expression was shut down in less than 4 h after induction, well before morphological changes were observed in these cells. At the enzymatic level, the activity of aspartate transcarbamylase, one of the three enzymes encoded by the CAD gene, decreased by about half within 24 h of induction, suggesting a CAD protein half-life of 24 h in differentiating HL-60 cells. Nevertheless, this means that significant levels of aspartate transcarbamylase activity remained even after the cells have stopped proliferating. From the RNA data, it is clear that CAD gene expression is rapidly turned off as promyelocytes begin to terminally differentiate into macrophages and granulocytes. We suspect that the inactivation of the CAD gene in induced HL-60 cells is a consequence of the differentiating cells leaving the cell cycle and becoming nonproliferating.

scholarly journals Original Chemical Series of Pyrimidine Biosynthesis Inhibitors That Boost the Antiviral Interferon Response

2017 ◽  
Vol 61 (10) ◽  
Author(s):  
Marianne Lucas-Hourani ◽  
Daniel Dauzonne ◽  
Hélène Munier-Lehmann ◽  
Samira Khiar ◽  
Sébastien Nisole ◽  
...  
Keyword(s):  
Metabolic Pathway ◽  
Defense Mechanisms ◽  
De Novo ◽  
Pyrimidine Biosynthesis ◽  
Interferon Response ◽  
Type I ◽  
Innate Defense ◽  
New Class ◽  
Biosynthesis Inhibitors ◽  
Tightly Coupled

ABSTRACT De novo pyrimidine biosynthesis is a key metabolic pathway involved in multiple biosynthetic processes. Here, we identified an original series of 3-(1H-indol-3-yl)-2,3-dihydro-4H-furo[3,2-c]chromen-4-one derivatives as a new class of pyrimidine biosynthesis inhibitors formed by two edge-fused polycyclic moieties. We show that identified compounds exhibit broad-spectrum antiviral activity and immunostimulatory properties, in line with recent reports linking de novo pyrimidine biosynthesis with innate defense mechanisms against viruses. Most importantly, we establish that pyrimidine deprivation can amplify the production of both type I and type III interferons by cells stimulated with retinoic acid-inducible gene 1 (RIG-I) ligands. Altogether, our results further expand the current panel of pyrimidine biosynthesis inhibitors and illustrate how the production of antiviral interferons is tightly coupled to this metabolic pathway. Functional and structural similarities between this new chemical series and dicoumarol, which was reported before to inhibit pyrimidine biosynthesis at the dihydroorotate dehydrogenase (DHODH) step, are discussed.

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