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Zinc is thought to be required as a structural component of the thyroid hormone (triiodothyronine, T3) receptor (TR). However, we have previously demonstrated that use of diethylenetriaminepentaacetic acid (DTPA) to restrict zinc availability to cultured cells actually potentiates rather than inhibits thyroid hormone action. In this article, the mechanisms underlying these effects of DTPA have been investigated. Treatment of GH3 rat pituitary tumor cells with DTPA in the presence of T3 resulted in twofold greater concentrations of growth hormone (GH) mRNA. Addition of actinomycin D to inhibit transcription showed that GH mRNA was actually less stable in the presence of DTPA, eliminating mRNA stabilization as a possible mechanism underlying this effect. Cycloheximide was able to block the induction by DTPA, showing a requirement for protein synthesis. Transient transfection of a GH promoter/luciferase reporter construct into GH3 cells revealed an inhibitory effect of DTPA on luciferase activity. However, when cells were stably transfected with the same construct, a T3-dependent stimulation of luciferase activity by DTPA was observed, mimicking the effects seen with the endogenous mRNA. Thus, the GH promoter does mediate the effects of DTPA, but stable integration into chromosomal material is required.