Early effects of thyrotropin on ribonucleic acid transcription in the thyroid.

Previous studies on the effects of TSH on the incorporation of labeled precursors into RNA have demonstrated an increase in the percentage of label in the phosphorylated nucleotide precursors of RNA. To avoid this nonspecific effect of TSH, we chose to measure RNA transcription in a system which uses exogenous phosphorylated precursors. Isolated nuclei were prepared from the thyroids of dogs which had been injected 90 min earlier with TSH. TSH caused a mean increase of 168% above control in total transcriptional activity, 193% in polymerase II-mediated (alpha-amanitin-sensitive) activity, and 155% in RNA polymerase I- and III-mediated (alpha-amanitin-resistant) activity. To determine whether part of this early effect of TSH was due to increased activity of RNA polymerase, incubations were also carried out in the presence of actinomycin D, which blocks transcription of endogenous template but does not inhibit transcription of added synthetic template poly deoxyadenylic-deoxythymidylic acid (dA.dT). Under these conditions, about half of the TSH-induced increase in the transcription of endogenous template could still be detected. Thus part of the early effects of TSH on RNA synthesis appears to be mediated through increased polymerase activity as well as enhanced template activity.