Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1977 May 15;164(2):379–387. doi: 10.1042/bj1640379

Uridine kinase activities and pyrimidine nucleoside phosphorylation in fluoropyrimidine-sensitive and -resistant cell lines of the Novikoff hepatoma.

N Greenberg, D E Schumm, T E Webb
PMCID: PMC1164803  PMID: 195585

Abstract

Uridine kinase, the rate-limiting enzyme in the activation (phosphorylation) of uridine and the corresponding chemotherapeutic analogues, is present as two isoenzymes localized exclusively in the cytosol of rapidly growing neoplasms, including the S-37 sarcoma, EL-4 leukaemia, HeLa cells (a human carcinoma) and the Novikoff hepatoma. The activities of the isolated isoenzymes are markedly decreased when the concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations of ATP, phosphate or Mg2+ that are optimum in vitro are replaced by concentrations approximating to those found in vivo. Further, comparisons of the Km values of isolated uridine kinases with those for cellular uptake of pyrimidine nucleosides and their rate of intracellular phosphorylation suggest that nucleoside-transport systems play a rate-limiting role in nucleoside analogue activation and consequently that it is impossible to estimate the Km of uridine kinase in the intact cell. During the development of tumour-cell resistance to 5-fluorouracil or 5-fluorouridine in vivo there was an early differential increase in the activity of a low-affinity (high-Km) uridine kinase isoenzyme, as measured in cell extracts, and a 7-fold increase in the Km values for the uptake of both uridine and 5-fluorouridine into the intact resistant cells.

Full text

PDF

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ANDERSON E. P., BROCKMAN R. W. FEEDBACK INHIBITION OF URIDINE KINASE BY CYTIDINE TRIPHOSPHATE AND URIDINE TRIPHOSPHATE. Biochim Biophys Acta. 1964 Nov 15;91:380–386. doi: 10.1016/0926-6550(64)90067-2. [DOI] [PubMed] [Google Scholar]
  2. Almersjö O., Brandberg A., Gustavsson B. Concentration of biologically active 5-fluorouracil in general circulation during continuous portal infusion in man: a preliminary report. Cancer Lett. 1975 Nov;1(2):113–117. doi: 10.1016/s0304-3835(75)95831-0. [DOI] [PubMed] [Google Scholar]
  3. FISCHER G. A. Detective transport of amethopterin (methotrexate) as a mechanism of resistance to the antimetabolite in L5178Y leukemic cells. Biochem Pharmacol. 1962 Dec;11:1233–1234. doi: 10.1016/0006-2952(62)90200-9. [DOI] [PubMed] [Google Scholar]
  4. Freed J. J., Mezger-Freed L. Origin of thymidine kinase deficient (TK-) haploid frog cells via an intermediate thymidine transport deficient (TT-) phenotype. J Cell Physiol. 1973 Oct;82(2):199–212. doi: 10.1002/jcp.1040820208. [DOI] [PubMed] [Google Scholar]
  5. HEIDELBERGER C., GHOBAR A., BAKER R. K., MUKHERJEE K. L. Studies on fluorinated pyrimidines. X. In vivo studies on tumor resistance. Cancer Res. 1960 Jul;20:897–902. [PubMed] [Google Scholar]
  6. Jackson V., Chalkley R. The binding of estradiol-17 beta to the bovine endometrial nuclear membrane. J Biol Chem. 1974 Mar 10;249(5):1615–1626. [PubMed] [Google Scholar]
  7. Keefer R. C., MacNamara D. J., Schumm D. E., Billmire D. F., Webb T. E. Early temporal changes in the uridine kinase isozyme profile of the Novikoff hepatoma in response to 5-azacytidine treatment. Biochem Pharmacol. 1975 Jul 15;24(13-14):1287–1290. doi: 10.1016/0006-2952(75)90338-x. [DOI] [PubMed] [Google Scholar]
  8. Keefer R. C., Morris H. P., Webb T. E. 5-azacytidine-modified patterns of uridine kinase activities in normal and neoplastic tissues. Cancer Res. 1974 Sep;34(9):2260–2265. [PubMed] [Google Scholar]
  9. Klubes P., Cerna I. Effects of 5-fluorouracil on drug-metabolizing enzymes in the rat. Cancer Res. 1974 May;34(5):927–931. [PubMed] [Google Scholar]
  10. Krystal G., Scholefield P. G. The partial purification and properties of uridine kinase from Ehrlich ascites tumor cells. Can J Biochem. 1973 Apr;51(4):379–389. doi: 10.1139/o73-044. [DOI] [PubMed] [Google Scholar]
  11. Krystal G., Webb T. E. Multiple forms of uridine kinase in normal and neoplastic rat liver. Biochem J. 1971 Oct;124(5):943–947. doi: 10.1042/bj1240943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lee T., Karon M., Monparler R. L. Cellular phosphorylation of 1-beta-D-arabinofuranosylcytosine 5-azacytidine with intact fibrosarcoma and leukemic cells. Cancer Res. 1975 Sep;35(9):2506–2510. [PubMed] [Google Scholar]
  13. Levinson C. Steady-state distribution of phosphate across the membrane of the Ehrlich ascites tumor cell. Biochim Biophys Acta. 1970 Apr 21;203(2):317–325. doi: 10.1016/0005-2736(70)90146-x. [DOI] [PubMed] [Google Scholar]
  14. Li C. C., Hochstadt J. Membrane-associated enzymes involved in nucleoside processing by plasma membrane vesicles isolated from L929 cells grown in defined medium. J Biol Chem. 1976 Feb 25;251(4):1181–1187. [PubMed] [Google Scholar]
  15. Plagemann P. G. Nucleotide pools of Novikoff rat hepatoma cells growing in suspension culture. I. Kinetics of incorporation of nucleosides into nucleotide pools and pool sizes during growth cycle. J Cell Physiol. 1971 Apr;77(2):213–240. doi: 10.1002/jcp.1040770212. [DOI] [PubMed] [Google Scholar]
  16. REICHARD P., SKOLD O. Enzymes of uracil metabolism in the Ehrlich ascites tumour and mammalian liver. Biochim Biophys Acta. 1958 May;28(2):376–385. doi: 10.1016/0006-3002(58)90485-2. [DOI] [PubMed] [Google Scholar]
  17. REICHARD P., SKOLD O., KLEIN G. Possible enzymic mechanism for the development of resistance against fluorouracil in ascites tumours. Nature. 1959 Apr 4;183(4666):939–941. doi: 10.1038/183939a0. [DOI] [PubMed] [Google Scholar]
  18. REICHARD P., SKOLD O., KLEIN G., REVESZ L., MAGNUSSON P. H. Studies on resistance against 5-fluorouracil. I. Enzymes of the uracil pathway during development of resistance. Cancer Res. 1962 Feb;22:235–243. [PubMed] [Google Scholar]
  19. SKOELD O. STUDIES ON RESISTANCE AGAINST 5-FLUOROURACIL. IV. EVIDENCE FOR AN ALTERED URIDINE KINASE IN RESISTANT CELLS. Biochim Biophys Acta. 1963 Sep 17;76:160–162. doi: 10.1016/0006-3002(63)90025-8. [DOI] [PubMed] [Google Scholar]
  20. Stambrook P. J., Sisken J. E., Ebert J. D. Uridine uptake and its intracellular phosphorylation during the cell cycle. J Cell Physiol. 1973 Oct;82(2):267–275. doi: 10.1002/jcp.1040820215. [DOI] [PubMed] [Google Scholar]
  21. Wilkinson D. S., Tlsty T. D., Hanas R. J. The inhibition of ribosomal RNA synthesis and maturation in Novikoff hepatoma cells by 5-fluorouridine. Cancer Res. 1975 Nov;35(11 Pt 1):3014–3020. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES