Biochemical phenotype of 5-fluorouracil-resistant murine T-lymphoblasts with genetically altered CTP synthetase activity

B. Aronow, T. Watts, J. Lassetter, W. Washtien, Buddy Ullman

Research output: Contribution to journalArticle

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Abstract

From wild type mouse lymphoma cells, a clone, (FURT-1A) was isolated by virtue of its resistance to 1 μM 5-fluorouracil and 10 μM thymidine. In comparative growth rate experiments, FURT-1A cells were also less sensitive than parental cells to the growth inhibitory effects of thymidine, deoxyguanosine, 5-fluorouridine, and arabinosylcytosine. The altered growth sensitivity of FURT-1A cells to cytotoxic nucleosides was directly related to their decreased ability to accumulate the corresponding triphosphate from exogenous nucleoside. FURT-1A cells contained elevated cytidylate nucleotide pools which prevented normal growth sensitivity and interfered with the salvage of nucleosides by inhibiting nucleoside kinase activities, by stimulating nucleotide dephosphorylating activities, and by overcoming certain allosteric inhibitions imposed on ribonucleotide reductase. Metabolic flux experiments with [3H]uridine in situ indicated that FURT-1A cells had a 2-fold enhanced rate of conversion of UTP to CTP. Kinetic analyses indicated that the CTP synthetase activity in extracts of FURT-1A cells was refractory to inhibition by CTP. The genetic loss of normal allosteric inhibition of the CTP synthetase activity in FURT-1A cells could account for the unusual phenotypic properties of these cells and conferred a high spontaneous mutator phenotype to cells possessing this mutation.

Original languageEnglish (US)
Pages (from-to)9035-9043
Number of pages9
JournalJournal of Biological Chemistry
Volume259
Issue number14
StatePublished - 1984
Externally publishedYes

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CTP synthetase
Nucleosides
Fluorouracil
Cytidine Triphosphate
nucleoside phosphotransferase
Phenotype
Thymidine
Nucleotides
Ribonucleotide Reductases
Salvaging
Uridine Triphosphate
Uridine
Cytarabine
Refractory materials
Experiments
Growth
Fluxes
Kinetics
Deoxyguanosine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Biochemical phenotype of 5-fluorouracil-resistant murine T-lymphoblasts with genetically altered CTP synthetase activity. / Aronow, B.; Watts, T.; Lassetter, J.; Washtien, W.; Ullman, Buddy.

In: Journal of Biological Chemistry, Vol. 259, No. 14, 1984, p. 9035-9043.

Research output: Contribution to journalArticle

Aronow, B, Watts, T, Lassetter, J, Washtien, W & Ullman, B 1984, 'Biochemical phenotype of 5-fluorouracil-resistant murine T-lymphoblasts with genetically altered CTP synthetase activity', Journal of Biological Chemistry, vol. 259, no. 14, pp. 9035-9043.
Aronow, B. ; Watts, T. ; Lassetter, J. ; Washtien, W. ; Ullman, Buddy. / Biochemical phenotype of 5-fluorouracil-resistant murine T-lymphoblasts with genetically altered CTP synthetase activity. In: Journal of Biological Chemistry. 1984 ; Vol. 259, No. 14. pp. 9035-9043.
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