Pathways of pyrimidine salvage in Pseudomonas and former Pseudomonas: detection of recycling enzymes using high-performance liquid chromatography.

Pyrimidine salvage pathways are vital for all bacteria in that they share in the synthesis of RNA with the biosynthetic pathway in pyrimidine prototrophs, while supplying all pyrimidine requirements in pyrimidine auxotrophs. Salvage enzymes that constitute the pyrimidine salvage pathways were studied in 13 members of Pseudomonas and former pseudomonads. Because it has been established that all Pseudomonas lack the enzyme uridine/cytidine kinase (Udk) and all contain uracil phosphoribosyl transferase (Upp), these two enzymes were not included in this experimental work.

Regulation of the pyrimidine biosynthetic pathway in a pyrD knockout mutant of Pseudomonas aeruginosa.

In research to date, regulation of the pyrimidine biosynthetic pathway at the level of gene expression has not been shown for wild type Pseudomonas aeruginosa. No repression was observed when uracil was added to the growth medium nor was any derepression seen when Pyr(-) auxotrophs were limited for pyrimidines. Here we show that the addition of uracil to Pseudomonas minimal medium influenced the synthesis of pyrimidine enzymes, while starvation of a pyrimidine knockout mutant (pyrD) elicited derepression of the pyrimidine enzymes.

Pathways of pyrimidine salvage in Streptomyces.

Using 5-fluoropyrimidine analogues, high-performance liquid chromatography (HPLC), and the feeding of pyrimidine compounds to pyrimidine auxotrophs, the pathways for salvage of exogenous pyrimidine nucleosides and bases in Streptomyces were established. Selection for resistance to the analogues resulted in the isolation of strains of S. griseus lacking the following enzyme activities: uracil phosphoribosyltransferase (upp) and cytidine deaminase (cdd).

Pseudomonas aeruginosa dihydroorotases: a tale of three pyrCs.

Pseudomonas aeruginosa PAO1 was shown to contain three pyrC sequences. Two of these genes, designated pyrC (PA3527) and pyrC2 (PA5541), encode polypeptides with dihydroorotase (DHOase) activity, while the third, pyrC' (PA0401), encodes a DHOase-like polypeptide that lacks DHOase activity, but is necessary for the structure and function of ATCase. Both pyrC and pyrC2 were cloned and complemented an Escherichia coli pyrC mutant.