Control of the pyrimidine biosynthetic pathway in Pseudomonas pseudoalcaligenes.
The five de novo enzyme activities unique to the pyrimidine biosynthetic pathway were found to be present in Pseudomonas pseudoalcaligenes ATCC 17440. A mutant strain with 31-fold reduced orotate phosphoribosyltransferase (encoded by pyrE) activity was isolated that exhibited a pyrimidine requirement for uracil or cytosine. Uptake of the nucleosides uridine or cytidine by wild-type or mutant cells was not detectable; explaining the inability of the mutant strain to utilize either nucleoside to satisfy its pyrimidine requirement. When the wild-type strain was grown in the presence of uracil, the activities of the five de novo enzymes were depressed. Pyrimidine limitation of the mutant strain led to the increase in aspartate transcarbamoylase and dihydroorotate dehydrogenase activities by more than 3-fold, and dihydroorotase and orotidine 5'-monophosphate decarboxylase activities about 1.5-fold, as compared to growth with excess uracil. It appeared that the syntheses of the de novo enzymes were regulated by pyrimidines. In vitro regulation of aspartate transcarbamoylase activity in P. pseudoalcaligenes ATCC 17440 was investigated using saturating substrate concentrations; transcarbamoylase activity was inhibited by Pi, PPi, uridine ribonucleotides, ADP, ATP, GDP, GTP, CDP, and CTP.
West
T P
TP
Department of Chemistry, South Dakota State University, Brookings 57007.
D001221:Aspartate Carbamoyltransferase / Q000378:metabolism*; D004080:Dihydroorotase / Q000378:metabolism; D000090722:Dihydroorotate Dehydrogenase; D009962:Orotate Phosphoribosyltransferase / Q000378:metabolism; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D044925:Oxidoreductases Acting on CH-CH Group Donors; D011549:Pseudomonas / Q000201:enzymology / Q000378:metabolism*; D011743:Pyrimidines / Q000096:biosynthesis*; D013997:Time Factors
D000090722:Dihydroorotate Dehydrogenase; D011743:Pyrimidines; D010088:Oxidoreductases; D044925:Oxidoreductases Acting on CH-CH Group Donors; D001221:Aspartate Carbamoyltransferase; D009962:Orotate Phosphoribosyltransferase; D004080:Dihydroorotase; D009964:Orotidine-5'-Phosphate Decarboxylase; C030986:pyrimidine