Thyroid hormone induces beta1-adrenergic receptor gene transcription through a direct repeat separated by five...

Transcription of the rat gene for the beta1-adrenergic receptor (beta1-AR) is stimulated by thyroid hormone (T3) in ventricular myocytes. To identify the domains involved in the regulation of beta1-AR gene transcription by T3, three kb of 5'-flanking sequence of the rat beta1-AR gene were ligated to a luciferase reporter gene and transiently transfected into ventricular myocytes. By generating deletions in the rat beta1-AR promoter, a region between -125 and -100 was found to mediate a three-fold induction by T3. This element was able to confer T3 responsiveness to a neutral promoter driving the luciferase reporter gene. Through site directed mutagenesis of this region, it was determined that the T3 responsive element (TRE) was organized as a direct repeat separated by five nucleotides in which the 5'-most AGGTCG half-site was between nucleotides -105 to -102 and the 3'-most AGGTCA half-site between nucleotides -116 and -113. Both the thyroid hormone receptor isoforms alpha and beta bound to the oligomer representing the sequences between -125 and -100 most efficiently as heterodimers with the retinoid X receptor. This TRE is unusual in that it is a direct repeat separated by five nucleotides which is located 3' to the transcriptional start site.