Liganded T3 receptor β2 inhibits the positive feedback autoregulation of the gene for GATA2, a transcription factor critical for thyrotropin production


Autoři: Naoko Hirahara aff001;  Hiroko Misawa Nakamura aff002;  Shigekazu Sasaki aff002;  Akio Matsushita aff002;  Kenji Ohba aff003;  Go Kuroda aff002;  Yuki Sakai aff002;  Shinsuke Shinkai aff002;  Hiroshi Haeno aff004;  Takuhiro Nishio aff005;  Shuichi Yoshida aff005;  Yutaka Oki aff006;  Takafumi Suda aff002
Působiště autorů: Division of Endocrinology and Metabolism, Department of Internal medicine, Japanese Red Cross Shizuoka Hospital, Shizuoka, Shizuoka, Japan aff001;  Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan aff002;  Medical Education Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan aff003;  Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, The University of Tokyo Kashiwa, Kashiwa, Chiba, Japan aff004;  Department of Integrated Human Sciences, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan aff005;  Department of Family and Community Medicine, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan aff006
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0227646

Souhrn

The serum concentration of thyrotropin (thyroid stimulating hormone, TSH) is drastically reduced by small increase in the levels of thyroid hormones (T3 and its prohormone, T4); however, the mechanism underlying this relationship is unknown. TSH consists of the chorionic gonadotropin α (CGA) and the β chain (TSHβ). The expression of both peptides is induced by the transcription factor GATA2, a determinant of the thyrotroph and gonadotroph differentiation in the pituitary. We previously reported that the liganded T3 receptor (TR) inhibits transactivation activity of GATA2 via a tethering mechanism and proposed that this mechanism, but not binding of TR with a negative T3-responsive element, is the basis for the T3-dependent inhibition of the TSHβ and CGA genes. Multiple GATA-responsive elements (GATA-REs) also exist within the GATA2 gene itself and mediate the positive feedback autoregulation of this gene. To elucidate the effect of T3 on this non-linear regulation, we fused the GATA-REs at -3.9 kb or +9.5 kb of the GATA2 gene with the chloramphenicol acetyltransferase reporter gene harbored in its 1S-promoter. These constructs were co-transfected with the expression plasmids for GATA2 and the pituitary specific TR, TRβ2, into kidney-derived CV1 cells. We found that liganded TRβ2 represses the GATA2-induced transactivation of these reporter genes. Multi-dimensional input function theory revealed that liganded TRβ2 functions as a classical transcriptional repressor. Then, we investigated the effect of T3 on the endogenous expression of GATA2 protein and mRNA in the gonadotroph-derived LβT2 cells. In this cell line, T3 reduced GATA2 protein independently of the ubiquitin proteasome system. GATA2 mRNA was drastically suppressed by T3, the concentration of which corresponds to moderate hypothyroidism and euthyroidism. These results suggest that liganded TRβ2 inhibits the positive feedback autoregulation of the GATA2 gene; moreover this mechanism plays an important role in the potent reduction of TSH production by T3.

Klíčová slova:

DNA transcription – Gene expression – Gene regulation – Messenger RNA – Pituitary gland – Plasmid construction – Thyroid-stimulating hormone – Transcription factors


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