Role of ornithine decarboxylase on glycolytic enzyme induction during thymocyte proliferation.
Resting rat thymocytes were stimulated to cell cycle entry and proliferation by the addition of concanavalin A and interleukin 2 to the culture medium. Maximal rats of DNA and RNA synthesis and of glycolysis with a 20-fold increase in glucose uptake were observed 48 h after stimulation. Glycolytic enzyme levels increased 3-8-fold, also reaching their maxima 48 h after mitogenic stimulation. Actinomycin D (1.3-1.5 ng/ml) completely inhibited DNA and RNA synthesis in 24- and 48-h cultured cells but showed no inhibitory effect on glycolytic enzyme induction or on enhanced glycolysis. These results suggest the presence of sufficient mRNAs for the synthesis of glycolytic enzymes even though transcription is abolished by actinomycin D. Ornithine decarboxylase activity increased rapidly up to 41-fold within 6 h after stimulation with concanavalin A and interleukin 2. This rapid increase could not be prevented by actinomycin D; however, ornithine decarboxylase activity of thymocytes was affected after 24- and 48-h culture periods. Inhibition of ornithine decarboxylase by 5 mM difluoromethylornithine completely abolished glycolytic enzyme induction. This inhibitory effect could be reversed by exogeneous putrescine. The results obtained indicate the importance of early ornithine decarboxylase activation and polyamine biosynthesis for the induction of glycolytic enzymes in proliferating thymocytes.
D000818:Animals; D002453:Cell Cycle; D002478:Cells, Cultured; D000518:Eflornithine / Q000494:pharmacology; D004790:Enzyme Induction; D005260:Female; D006019:Glycolysis / Q000187:drug effects; D007700:Kinetics; D008213:Lymphocyte Activation; D008854:Microscopy, Electron; D009955:Ornithine Decarboxylase / Q000378:metabolism*; D011700:Putrescine / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013601:T-Lymphocytes / Q000166:cytology / Q000201:enzymology* / Q000276:immunology
D009955:Ornithine Decarboxylase; D011700:Putrescine; D000518:Eflornithine