Effect of actinomycin D on iodide transport in FRTL-5 thyroid cells.
TSH-induced I- uptake in FRTL-5 thyroid cells requires new protein synthesis. During the induction of I- uptake, which takes nearly 60 h to reach its maximum, two waves of protein synthesis can be identified: one during the first 8 h and another after 24-30 h, each involving different proteins. Cycloheximide (CH) added during the first 10 h of a 48-h incubation with TSH completely inhibits the induction of I- uptake; 58% inhibition is observed with CH added at 24 h; no inhibition is observed when CH is added 36 h after TSH. Like CH, actinomycin D (ActD), added at the beginning of the 48-h period, inhibits TSH induction of I- uptake; partial inhibition (83%, 72%, and 28%) is observed when ActD is added at 1, 5, and 10 h, respectively. Treatment with ActD at 24 h (ActD-treated cells), however, paradoxically increases I- uptake (1.8- to 3.5-fold over the control value). The characteristics of I- uptake in ActD-treated cells are the same as those in untreated cells; both are Na+ dependent and can be inhibited in a comparable manner by anions. Kinetic measurements of I- transport indicate that ActD increases the rate of I- influx (2-fold or greater increase in maximum velocity without a significant change in Km), with only minor changes in I- release. Enhanced I- uptake in ActD-treated cells is inhibited by the simultaneous (24-h) administration of CH, indicating that protein synthesis is required for the late ActD effect. Despite an overall 2-fold decrease in protein synthesis in cells treated with ActD at 24 h, the synthesis of individual proteins maximally induced by TSH during the first 8 h is increased, whereas that of some proteins maximally synthesized after 24-30 h is markedly reduced. The present data indicate that TSH-induced I- uptake in FRTL-5 cells involves a regulatory action of TSH that operates at the mRNA level.
D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology*; D007454:Iodides / Q000378:metabolism*; D051381:Rats; D013961:Thyroid Gland / Q000166:cytology / Q000378:metabolism*; D013972:Thyrotropin / Q000494:pharmacology; D013997:Time Factors
D007454:Iodides; D003609:Dactinomycin; D013972:Thyrotropin; D003513:Cycloheximide