Ribonucleotide reductase and thymidine phosphorylation: two potential targets of azodicarbonamide.
Azodicarbonamide tested as an anti-HIV agent was reported to expulse zinc from viral zinc-cysteine factors and to inhibit calcium mobilization machinery. It has structural analogy with hydroxyurea that inhibits ribonucleotide reductase and could also act on this target. Azodicarbonamide was therefore tested for its capacity to modulate deoxyribonucleotides triphosphate pools alone or in combination with other agents in the lymphoblastic SUP-T1 cell line susceptible to HIV infection. The deoxyribonucleotides triphosphate were evaluated by an enzymatic assay using sequenase. Two hours exposure of SUP-T1 cells to 100 microM azodicarbonamide induced a 50% reduction of each deoxyribonucleotide triphosphate. Among other inhibitors of nucleotide metabolism (hydroxyurea, methotrexate and thymidine), hydroxyurea only reproduces the effect of azodicarbonamide. This suggests, but does not demonstrate directly, that azodicarbonamide inhibits ribonucleotide reductase activity. The combination of azodicarbonamide with each of these inhibitors affected particularly the dCTP pool. During this study it was also suggested that azodicarbonamide could interfere with thymidine phosphorylation. Thymidine phosphorylating activity was measured with 3H-thymidine as substrate. In acellular preparations, azodicarbonamide also non-competitively inhibits thymidine phosphorylating activity. This effect was not reproduced by hydroxyurea. Thus, in vitro azodicarbonamide decreases the intracellular pool of deoxyribonucleotide and thymidine phosphorylation.
Fagny
Christine
C
Department of Biochemistry and Nutrition, Faculty of Medicine, Université Libre de Bruxelles, Brussels, Belgium.
Vandevelde
Michel
M
Svoboda
Michal
M
Robberecht
Patrick
P
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001391:Azo Compounds / Q000494:pharmacology*; D003845:Deoxycytosine Nucleotides / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism; D004347:Drug Interactions; D006918:Hydroxyurea / Q000494:pharmacology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008727:Methotrexate / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D012264:Ribonucleotide Reductases / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism* / Q000494:pharmacology; D013942:Thymine Nucleotides / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D001391:Azo Compounds; D003845:Deoxycytosine Nucleotides; D003848:Deoxyguanine Nucleotides; D007166:Immunosuppressive Agents; D013942:Thymine Nucleotides; C024107:2'-deoxycytidine 5'-triphosphate; C004525:azodicarbonamide; C029603:deoxyguanosine triphosphate; D012264:Ribonucleotide Reductases; C024157:thymidine 5'-triphosphate; D013936:Thymidine; D006918:Hydroxyurea; D008727:Methotrexate