Abnormalities in uridine homeostatic regulation and pyrimidine nucleotide metabolism as a consequence of the deletion of the uridine phosphorylase gene.
We report in the present study the critical role of uridine phosphorylase (UPase) in uridine homeostatic regulation and pyrimidine nucleotide metabolism, employing newly developed UPase-/- mice. Our data demonstrate that the abrogation of UPase activity led to greater than a 6-fold increase in uridine concentrations in plasma, a 5-6-fold increase in lung and gut, and a 2-3-fold increase in liver and kidney, as compared with wild type mice. Urine uridine levels increased 24-fold normal in UPase-/- mice. Uridine half-life and the plasma retention of pharmacological doses of uridine were significantly prolonged. Further, in these UPase-/- mice, abnormal uridine metabolism led to disorders of various nucleotide metabolisms. In the liver, gut, kidney, and lung of UPase-/- mice, total uridine ribonucleotide concentrations increased 2-3 times as compared with control mice. Cytidine ribonucleotides and adenosine and guanosine ribonucleotides also increased, although to a lesser extent, in these organs. Most significant deoxyribonucleotide changes were present in the gut and lung of UPase-/- mice. In these tissues, dTTP concentration increased more than 4-fold normal, and dCTP, dGTP, and dATP concentrations rose 1-2 times normal. In kidney, dTTP concentration increased 2-fold normal, and dCTP and dGTP concentrations rose less than 1-fold normal. In addition, the accumulated uridine in plasma and tissues efficiently reduced 5-fluorouracil host toxicity and altered the anesthetic effect of pentobarbital. These data indicate that UPase is a critical enzyme in the regulation of uridine homeostasis and pyrimidine nucleotide metabolism, and 5-fluorouracil activity.
Cao
Deliang
D
Cancer Institute, Southern Illinois University School of Medicine, Springfield, Illinois 62702, USA.
Leffert
Janine J
JJ
McCabe
James
J
Kim
Bradford
B
Pizzorno
Giuseppe
G
D000241:Adenosine / Q000737:chemistry; D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology; D015139:Blotting, Southern; D003562:Cytidine / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000737:chemistry; D005472:Fluorouracil / Q000494:pharmacology; D017353:Gene Deletion; D006151:Guanosine / Q000737:chemistry; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D009711:Nucleotides / Q000737:chemistry; D010424:Pentobarbital / Q000737:chemistry / Q000494:pharmacology; D011743:Pyrimidines / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012265:Ribonucleotides / Q000737:chemistry; D013942:Thymine Nucleotides / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution; D014529:Uridine / Q000737:chemistry* / Q000378:metabolism; D014543:Uridine Phosphorylase / Q000378:metabolism*
D000964:Antimetabolites, Antineoplastic; D003854:Deoxyribonucleotides; D009711:Nucleotides; D011743:Pyrimidines; D012333:RNA, Messenger; D012265:Ribonucleotides; D013942:Thymine Nucleotides; D006151:Guanosine; D003562:Cytidine; D014543:Uridine Phosphorylase; D004259:DNA-Directed DNA Polymerase; D010424:Pentobarbital; D000241:Adenosine; C030986:pyrimidine; C024157:thymidine 5'-triphosphate; D005472:Fluorouracil; D014529:Uridine