Purine catabolism in Escherichia coli and function of xanthine dehydrogenase in purine salvage.
Escherichia coli is not known to utilize purines, other than adenine and adenosine, as nitrogen sources. We reinvestigated purine catabolism because a computer analysis suggested several potential sigma(54)-dependent promoters within a 23-gene cluster whose products have homology to purine catabolic enzymes. Our results did not provide conclusive evidence that the sigma(54)-dependent promoters are active. Nonetheless, our results suggest that some of the genes are metabolically significant. We found that even though several purines did not support growth as the sole nitrogen source, they did stimulate growth with aspartate as the nitrogen source. Cells produced (14)CO(2) from minimal medium containing [(14)C]adenine, which implies allantoin production. However, neither ammonia nor carbamoyl phosphate was produced, which implies that purine catabolism is incomplete and does not provide nitrogen during nitrogen-limited growth. We constructed strains with deletions of two genes whose products might catalyze the first reaction of purine catabolism. Deletion of one eliminated (14)CO(2) production from [(14)C]adenine, which implies that its product is necessary for xanthine dehydrogenase activity. We changed the name of this gene to xdhA. The xdhA mutant grew faster with aspartate as a nitrogen source. The mutant also exhibited sensitivity to adenine, which guanosine partially reversed. Adenine sensitivity has been previously associated with defective purine salvage resulting from impaired synthesis of guanine nucleotides from adenine. We propose that xanthine dehydrogenase contributes to this purine interconversion.
Xi
H
H
Department of Molecular and Cell Biology, The University of Texas at Dallas, Richardson, Texas 75083-0688, USA.
Schneider
B L
BL
Reitzer
L
L
D000225:Adenine / Q000494:pharmacology; D002245:Carbon Dioxide / Q000378:metabolism; D003470:Culture Media; D004268:DNA-Binding Proteins; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D009584:Nitrogen / Q000378:metabolism; D010641:Phenotype; D011401:Promoter Regions, Genetic; D011687:Purines / Q000378:metabolism*; D051877:RNA Polymerase Sigma 54; D012808:Sigma Factor / Q000378:metabolism; D014968:Xanthine Dehydrogenase / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D011687:Purines; D012808:Sigma Factor; C497751:rpoN protein, E coli; D002245:Carbon Dioxide; D014968:Xanthine Dehydrogenase; D012321:DNA-Directed RNA Polymerases; D051877:RNA Polymerase Sigma 54; D000225:Adenine; D009584:Nitrogen; C030985:purine