title: GATA2 mediates thyrotropin-releasing hormone-induced transcriptional activation of the thyrotropin β gene.
abstract: Thyrotropin-releasing hormone (TRH) activates not only the secretion of thyrotropin (TSH) but also the transcription of TSHβ and α-glycoprotein (αGSU) subunit genes. TSHβ expression is maintained by two transcription factors, Pit1 and GATA2, and is negatively regulated by thyroid hormone (T3). Our prior studies suggest that the main activator of the TSHβ gene is GATA2, not Pit1 or unliganded T3 receptor (TR). In previous studies on the mechanism of TRH-induced activation of the TSHβ gene, the involvements of Pit1 and TR have been investigated, but the role of GATA2 has not been clarified. Using kidney-derived CV1 cells and pituitary-derived GH3 and TαT1 cells, we demonstrate here that TRH signaling enhances GATA2-dependent activation of the TSHβ promoter and that TRH-induced activity is abolished by amino acid substitution in the GATA2-Zn finger domain or mutation of GATA-responsive element in the TSHβ gene. In CV1 cells transfected with TRH receptor expression plasmid, GATA2-dependent transactivation of αGSU and endothelin-1 promoters was enhanced by TRH. In the gel shift assay, TRH signal potentiated the DNA-binding capacity of GATA2. While inhibition by T3 is dominant over TRH-induced activation, unliganded TR or the putative negative T3-responsive element are not required for TRH-induced stimulation. Studies using GH3 cells showed that TRH-induced activity of the TSHβ promoter depends on protein kinase C but not the mitogen-activated protein kinase, suggesting that the signaling pathway is different from that in the prolactin gene. These results indicate that GATA2 is the principal mediator of the TRH signaling pathway in TSHβ expression.
authors.affiliation: Second Division, Department of Internal Medicine, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Ohba
authors.forename: Shigekazu
authors.initials: S
authors.lastname: Sasaki
authors.forename: Akio
authors.initials: A
authors.lastname: Matsushita
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Iwaki
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Matsunaga
authors.forename: Shingo
authors.initials: S
authors.lastname: Suzuki
authors.forename: Keiko
authors.initials: K
authors.lastname: Ishizuka
authors.forename: Hiroko
authors.initials: H
authors.lastname: Misawa
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Oki
authors.forename: Hirotoshi
authors.initials: H
authors.lastname: Nakamura
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D050989:GATA2 Transcription Factor / Q000502:physiology*; D006801:Humans; D009154:Mutation; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D037322:Thyrotropin, beta Subunit / Q000235:genetics*; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism / Q000502:physiology*; D015533:Transcriptional Activation / Q000502:physiology*
chemical_list: D017931:DNA Primers; D050989:GATA2 Transcription Factor; C494711:GATA2 protein, human; D037322:Thyrotropin, beta Subunit; D013973:Thyrotropin-Releasing Hormone