Activation of deoxycytidine kinase by inhibition of DNA synthesis in human lymphocytes

Z. Csapó, M. Sasvári, T. Spasokoukotskaja, Iannis Talianidis, Staffan Eriksson, M. Staub

Research output: Article

41 Citations (Scopus)

Abstract

Deoxycytidine kinase (dCK, EC.2.7.1.74) is a key enzyme in the intracellular metabolism of 2-chlorodeoxyadenosine, 1β-D-arabinofuranosylcytosine, difluorodeoxycytidine, and other drugs used in chemotherapy of different leukaemias and solid tumours. Recently, stimulation of dCK activity was shown by these analogues and by other genotoxic agents such as etoposide and NaF, all of which cause severe inhibition of DNA synthesis in cell cultures. Here we describe that direct inhibition of DNA polymerases by aphidicolin stimulated dCK activity in normal lymphocytes and acute myeloid leukaemic cells, as well as in HL 60 promyelocytic cell cultures. Increased dCK activity was not due to new protein synthesis under our conditions, as measured by immunoblotting. Partial purification by diethylaminoethyl-Sephadex chromatography revealed that the activated form of dCK survived purification procedure. Moreover, it was possible to inactivate purified dCK preparations by recombinant protein phosphatase with Ser/Thr/Tyr dephosphorylating activity. These data suggest that the activation of dCK may be due to phosphorylation, and that deoxynucleoside salvage is promoted during inhibition of DNA synthesis in human lymphocytes.

Original languageEnglish
Pages (from-to)191-197
Number of pages7
JournalBiochemical Pharmacology
Volume61
Issue number2
DOIs
Publication statusPublished - jan. 15 2001

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Deoxycytidine Kinase
Lymphocytes
Cell Culture Techniques
Chemical activation
Cladribine
Aphidicolin
Cell culture
Purification
HL-60 Cells
Phosphoprotein Phosphatases
DNA
Cytarabine
Etoposide
Myeloid Cells
DNA-Directed DNA Polymerase
Recombinant Proteins
Immunoblotting
Chromatography
Salvaging
Leukemia

ASJC Scopus subject areas

  • Pharmacology

Cite this

Activation of deoxycytidine kinase by inhibition of DNA synthesis in human lymphocytes. / Csapó, Z.; Sasvári, M.; Spasokoukotskaja, T.; Talianidis, Iannis; Eriksson, Staffan; Staub, M.

In: Biochemical Pharmacology, Vol. 61, No. 2, 15.01.2001, p. 191-197.

Research output: Article

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