Polyamine regulation of the synthesis of thymidine kinase in bovine lymphocytes.
Concanavalin A-activated lymphocytes were made polyamine deficient by treatment with alpha-difluoromethylornithine and ethylglyoxal bis(guanylhydrazone). Thymidine kinase activity in polyamine-deficient cells was 17% of the level in normal cells. Thymidine kinase mRNA increased with time after concanavalin A activation and reached a maximum at 36 h after concanavalin A addition. The amount of thymidine kinase mRNA in polyamine-deficient cells was approximately 75% of that in normal cells. The transcription of thymidine kinase gene in isolated nuclei of polyamine-deficient cells was also 75% of that from normal cells. The turnover rate of thymidine kinase mRNA in both normal and polyamine-deficient cells was nearly equal. In normal cells, 95% of thymidine kinase mRNA was polysome associated, while in polyamine-deficient cells, 60% of the mRNA was polysome associated. In addition, the size of polysomes associated with thymidine kinase mRNA in polyamine-deficient cells was smaller than that in normal cells. Synthesis of thymidine kinase was stimulated approximately seven-fold by 0.3 mM spermidine in a rabbit reticulocyte polyamine-free protein synthetic system. The half-life of thymidine kinase activity in both normal and polyamine-deficient cells was nearly equal. Thymidine kinase activity was not influenced significantly by 0.3 mM spermidine. These combined results suggested that the synthesis of thymidine kinase was mainly regulated by polyamines at the level of translation.
Ito
K
K
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Igarashi
K
K
D012437:Adenosylmethionine Decarboxylase / Q000037:antagonists & inhibitors; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002478:Cells, Cultured; D003208:Concanavalin A; D003513:Cycloheximide / Q000494:pharmacology; D004257:DNA Polymerase II / Q000378:metabolism; D015342:DNA Probes; D004261:DNA Replication / Q000187:drug effects; D000518:Eflornithine / Q000494:pharmacology*; D007700:Kinetics; D008213:Lymphocyte Activation; D008214:Lymphocytes / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D008935:Mitoguazone / Q000031:analogs & derivatives* / Q000494:pharmacology; D008969:Molecular Sequence Data; D011073:Polyamines / Q000378:metabolism*; D011132:Polyribosomes / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics; D013095:Spermidine / Q000494:pharmacology*; D013937:Thymidine Kinase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D013940:Thymidylate Synthase / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*
D015342:DNA Probes; D011073:Polyamines; D012333:RNA, Messenger; D003208:Concanavalin A; C041127:ethylglyoxal bis(guanylhydrazone); D003513:Cycloheximide; D013940:Thymidylate Synthase; D013937:Thymidine Kinase; D004257:DNA Polymerase II; D012437:Adenosylmethionine Decarboxylase; D008935:Mitoguazone; D013095:Spermidine; D000518:Eflornithine