Evaluation of thyroid hormone effects on liver P450 reductase translation

The expression of NADPH cytochrome P450 oxidoreductase (P450R) in rat liver is positively regulated by thyroid hormone (T3), at both the transcriptional and post-transcriptional levels. Here we investigate the effects of T3-induced hyperthyroidism on the regulation of P450R protein synthesis. T3 treatment of adult male rats led to a strong induction (up to ∼10-fold) of liver P450R mRNA but little or no change in P450R protein and activity. Investigation of this discrepancy revealed that the association of hepatic P450R mRNA with polysomes was not altered by T3 treatment, suggesting that the discoordinate changes in P450R mRNA and protein levels do not reflect decreased recruitment of T3-induced P450R mRNA into polysomes. Moreover, polysome size distribution analysis of P450R mRNA did not show any T3-dependent changes. When assayed in an in vitro translation system, T3-induced and uninduced P450R mRNAs were translated with similar efficiencies. Moreover, liver cell extract from T3-treated rats did not selectively inhibit in vitro translation of T3-induced P450R mRNA. Thus, neither structural changes in P450R mRNA nor trans-acting binding proteins in liver cytosol were found to control translation of P450R mRNA in response to T3 treatment. Taken together, these data suggest that P450R may in part be regulated at the level of protein stability in hyperthyroid rat liver.