Distinct and Histone-Specific Modifications Mediate Positive versus Negative Transcriptional Regulation of TSHα Promoter

Background Hormonally-regulated histone modifications that govern positive versus negative transcription of target genes are poorly characterized despite their importance for normal and pathological endocrine function. There have been only a few studies examining chromatin modifications on target gene promoters by nuclear hormone receptors. Moreover, these studies have focused on positively-regulated target genes. TSHα, a heterodimer partner for thyrotropin (TSH), is secreted by the pituitary gland. T 3 negatively regulates TSHα gene expression via thyroid hormone receptors (TRs) which belong to the nuclear hormone receptor superfamily, whereas thyrotropin releasing hormone (TRH) positively regulates via the TRH receptor, a G protein-coupled receptor. Methodology/Principal Findings We studied regulation of the TSHα gene by cAMP and T 3 using chromatin immunoprecipitation (ChIP) assays in stably-transfected rat pituitary cells containing the human TSHα promoter. Interestingly, cAMP selectively increased histone H4 acetylation whereas, as previously reported, T 3 induced histone H3 acetylation. In particular, cAMP increased H4K5 and H4K8 acetylation and decreased H4K20 trimethylation, modifications associated with transcriptional activation. T 3 increased H3K9 and H3K18 acetylation and H3K4 trimethylation; however, it also decreased H3K27 acetylation and increased H3K27 trimethylation which are associated with transcriptional repression. Of note, cAMP recruited pCREB, CBP/p300, and PCAF to the promoter whereas T 3 caused dissociation of NCoR/SMRT and HDAC3. Overexpression of a dominant negative mutant thyroid hormone receptor (TR) from a patient with resistance to thyroid hormone (RTH) led to less T 3 -dependent negative regulation and partially blocked histone H3 modifications of the TSHα promoter. Conclusions/Significance Our findings show that non-overlapping and specific histone modifications determine positive versus negative transcriptional regulation, and integrate opposing hormonal and intracellular signals at the TSHα promoter. A mutant TR from a patient with RTH exerted dominant negative activity by blocking the histone modifications induced by T 3 on the TSHα promoter and likely contributes to the inappropriate TSH production observed in RTH.