Cycloheximide enhances ACTH-receptor mRNA through transcriptional and post-transcriptional mechanisms in bovine adrenocortical cells

Abstract

We have previously shown that ACTH is one of the few polypeptide hormones having a positive trophic effect, not only on the number, but also on the expression of its own receptors. In the present study, we investigated whether the constitutive and ACTH-induced expression of ACTH-receptor (ACTH-R) mRNA in bovine adrenocortical cells (BAC) requires new protein synthesis. The results show that cycloheximide alone, an inhibitor of protein synthesis, induced a time- and dose-dependent increase in the constitutive level of the ACTH-R major transcript of 3.6 kb in BAC. The maximal stimulation (5.17 ± 1.15 fold, n = 4) was obtained after 24 h treatment with 5 μg/ml cycloheximide. The effect of cycloheximide was specific and not directly related to translational arrest since other protein synthesis inhibitors acting through different mechanisms, emetine and puromycin, were unable to reproduce such an effect at concentrations inhibiting protein synthesis. The effect of cycloheximide involved an increase in the half-life and the transcription rate of the major transcript of ACTH-R (2- and 8.4-fold respectively). In addition, the results also demonstrated that neither the constitutive nor the ACTH-induced expression of ACTH-R require new protein synthesis.