Activation of deoxycytidine kinase by inhibition of DNA synthesis in human lymphocytes.
Deoxycytidine kinase (dCK, EC.2.7.1.74) is a key enzyme in the intracellular metabolism of 2-chlorodeoxyadenosine, 1-beta-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.
Csapó
Z
Z
Department of Medical Chemistry, Molecular Biology and Pathobiochemistry, Semmelweis University, H-1444 8, Budapest, Hungary.
Sasvári-Székely
M
M
Spasokoukotskaja
T
T
Talianidis
I
I
Eriksson
S
S
Staub
M
M
D016590:Aphidicolin / Q000494:pharmacology*; D004247:DNA / Q000096:biosynthesis / Q000187:drug effects*; D003842:Deoxycytidine Kinase / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D018922:HL-60 Cells; D006801:Humans; D008214:Lymphocytes / Q000187:drug effects* / Q000201:enzymology; D019384:Nucleic Acid Synthesis Inhibitors; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D010766:Phosphorylation
D004791:Enzyme Inhibitors; D019384:Nucleic Acid Synthesis Inhibitors; D016590:Aphidicolin; D004247:DNA; D003842:Deoxycytidine Kinase; D010749:Phosphoprotein Phosphatases