The molecular mechanism for hormone- and serum-dependent regulation of thyroglobulin (TG) gene expression was studied. A construct of rat TG promoter (−178 to −3) linked to a luciferase gene was transfected into TSH-, insulin- and serum-deprived FRTL-5 cells. Addition of TSH, insulin or serum augmented the luciferase activity. The endogenous TG mRNA level was also increased, indicating that the promoter used confers responsiveness of TG gene to these additives. The possible involvement of thyroid-transcription factors, TTF-1, TTF-2 and Pax-8, in the induction of TG gene transcription was studied using an electrophoretic mobility shift assay. Since the protein/DNA ratio in FRTL-5 cell extracts was significantly increased by these additives, binding activities of these factors per unit of DNA were examined. It was demonstrated that TSH, insulin or serum increased not only TTF-2 binding activity but also the binding activities of TTF-1 and Pax-8. However, the magnitude of the increase in TTF-1 and Pax-8 mRNA levels per unit of DNA was less than that of the binding activity. Taken together, our results suggest that TSH, insulin and serum increase the binding activities of TTF-1 and Pax-8 to the TG promoter presumably through the posttranslational modification of the factors, thereby enhancing TG gene transcription.
Journal of Endocrinology (1996) 150, 287–298
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