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Uracil salvage pathway in PC12 cells.

L Mascia1, G Turchi, V Bemi

  • 1Department of Physiology and Biochemistry, University of Pisa, Italy. lmascia@dfb.unipi.it

Biochimica Et Biophysica Acta
|November 18, 2000
PubMed
Summary

PC12 cells salvage uracil into nucleotides via a ribose 1-phosphate-dependent pathway, independent of pyrimidine base phosphoribosyl transferase. This finding is crucial for understanding chemotherapeutics

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Neuroscience

Background:

  • PC12 cells lack pyrimidine base phosphoribosyl transferase, necessitating alternative uracil salvage pathways.
  • Uracil salvage in these cells must occur independently of 5-phosphoribosyl 1-pyrophosphate.

Purpose of the Study:

  • To investigate the salvage anabolism of uracil in PC12 cells.
  • To elucidate the mechanism of uracil nucleotide synthesis in the absence of pyrimidine base phosphoribosyl transferase.
  • To determine the relevance of in vitro findings to intact PC12 cells.

Main Methods:

  • Incubation of PC12 cell extracts with ribose 1-phosphate, ATP, and uracil.
  • Analysis of intracellular metabolites in intact PC12 cells following uracil uptake.
  • Enzymatic assays to identify key enzymes in the salvage pathway.

Main Results:

  • PC12 cell extracts catalyze uracil nucleotide synthesis via a ribose 1-phosphate-dependent pathway involving uridine phosphorylase and subsequent uridine phosphorylation.
  • Intact PC12 cells readily uptake external uracil and salvage it into uracil nucleotides with uridine as an intermediate.
  • The identified salvage pathway is similar to that observed in rat liver and brain extracts.

Conclusions:

  • PC12 cells utilize a ribose 1-phosphate-dependent uracil salvage pathway, independent of 5-phosphoribosyl 1-pyrophosphate.
  • This pathway, involving uridine phosphorylase, is operative in both cell extracts and intact PC12 cells.
  • Understanding this pathway is critical for evaluating the impact of novel chemotherapeutics on pyrimidine metabolism in neurons.

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