Thyroid hormone inhibition of human thyrotropin beta-subunit gene expression is mediated by a cis-acting element located in the first exon.
Thyroid hormone regulation of the human thyrotropin beta-subunit gene (TSH beta) was examined in a human embryonal cell line (293). Transient expression studies were performed with chimeric plasmids containing the reporter gene, chloramphenicol acetyltransferase. Sequences in the first exon between +9 and +37 base pairs (bp) enhanced gene expression from the human TSH beta promoter in the absence of thyroid hormone as well as mediated a concentration-dependent triiodothyronine (L-T3) decrease in gene expression. Thyroid hormone inhibition of expression was also conferred to the herpes simplex virus thymidine kinase promoter by inserting +3 to +37 bp of the human TSH beta gene downstream from the start of transcription. Primer extension analysis of RNA from transfected cell cultures revealed accurate transcription initiation in only those constructs which contained sequences between +9 and +37 bp. Moreover, RNA analysis confirmed that L-T3 inhibition of chloramphenicol acetyltransferase activity from chimeric pTSH beta CAT constructs occurred at a pretranslational level. In addition, a nuclear thyroid hormone receptor, c-erbA-beta, bound to this region in an avidin-biotin DNA binding assay. These data suggest that L-T3, bound to its receptor, may inhibit human TSH beta expression by interfering with an element that functions to enhance gene expression.
Molecular, Cellular, and Nutritional Endocrinology Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.
F E
FE
Wondisford
E A
EA
Farr
S
S
Radovick
H J
HJ
Steinfelder
J M
JM
Moates
J H
JH
McClaskey
B D
BD
Weintraub
D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D005091:Exons; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D010957:Plasmids; D011988:Receptors, Thyroid Hormone / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000187:drug effects*; D013963:Thyroid Hormones / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013972:Thyrotropin / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics
D011988:Receptors, Thyroid Hormone; D013963:Thyroid Hormones; D014157:Transcription Factors; D013972:Thyrotropin; D015500:Chloramphenicol O-Acetyltransferase