Caffeine-DNA interactions: biochemical investigations comprising DNA-repair enzymes and nucleic acid synthesis.
Chicken embryo cells were treated with caffeine (0.5-8.0 mM) alone or combined with various chemical and physical DNA-and/or chromatin-interactive agents. Analytical procedures comprised scheduled (SDS) and unscheduled (UDS) DNA synthesis, RNA synthesis (RNS), the activities of O6-alkylguanine-DNA alkyltransferase (AT) and poly (ADP-ribose) polymerase (PARP) as well as nucleoid sedimentation. Additional investigations were done in rat thymic and splenic cells. The effect of caffeine on DNase-I activity served as an in vitro-model system. When present in the PARP-, SDS-, UDS- and RNS-assays, caffeine inhibited the corresponding tracer (14C-NAD, dT-3H, 3H-U) incorporation in a dose-dependent manner. The AT activity was slightly stimulated. At concentrations of 0.06-0.3 mM, caffeine inhibited DNase-I activity by excess substrate. No specific effects of caffeine could be shown by nucleoid sedimentation. Besides the reduced permeability of the cells to nucleic acid precursors, the results obtained with the PARP- and DNase-I assays give evidence for the formation of a DNA-caffeine adduct as a prominent mechanism of cellular caffeine effects including DNA repair inhibition.
Tempel
K
K
Institut für Pharmakologie, Toxikologie und Pharmazie, Universität München, Germany.
von Zallinger
C
C
D036002:ADP Ribose Transferases / Q000037:antagonists & inhibitors; D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000378:metabolism; D001549:Benzamides / Q000494:pharmacology; D001921:Brain / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002110:Caffeine / Q000494:pharmacology*; D002470:Cell Survival; D002642:Chick Embryo; D004247:DNA / Q000096:biosynthesis* / Q000187:drug effects; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004622:Embryo, Mammalian; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004996:Ethidium / Q000494:pharmacology; D005260:Female; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D012313:RNA / Q000096:biosynthesis / Q000187:drug effects; D051381:Rats; D017208:Rats, Wistar; D013601:T-Lymphocytes / Q000187:drug effects / Q000378:metabolism
D001549:Benzamides; D004791:Enzyme Inhibitors; D002110:Caffeine; D012313:RNA; C025160:3-aminobenzamide; D004247:DNA; D019853:O(6)-Methylguanine-DNA Methyltransferase; D036002:ADP Ribose Transferases; D011065:Poly(ADP-ribose) Polymerases; D003850:Deoxyribonuclease I; D004996:Ethidium