School of Biological Sciences and Technology, Chonnam National University, Gwangju 61186, Korea.
Laboratory Animal Resource Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Korea.
Int J Mol Sci. 2020 Sep 28;21(19):7148. doi: 10.3390/ijms21197148.
Bone morphogenetic protein 6 (BMP6) is a multifunctional growth factor involved in organ development and homeostasis. BMP6 controls expression of the liver hormone, hepcidin, and thereby plays a crucial role in regulating iron homeostasis. BMP6 gene transcriptional regulation in liver is largely unknown, but would be of great help to externally modulate iron load in pathologic conditions. Here, we describe a detailed molecular mechanism of hepatic BMP6 gene expression by an orphan nuclear receptor, estrogen-related receptor γ (ERRγ), in response to the pro-inflammatory cytokine interleukin 6 (IL-6). Recombinant IL-6 treatment increases hepatic ERRγ and BMP6 expression. Overexpression of ERRγ is sufficient to increase BMP6 gene expression in hepatocytes, suggesting that IL-6 is upstream of ERRγ. In line, knock-down of ERRγ in cell lines or a hepatocyte specific knock-out of ERRγ in mice significantly decreases IL-6 mediated BMP6 expression. Promoter studies show that ERRγ directly binds to the ERR response element (ERRE) in the mouse BMP6 gene promoter and positively regulates BMP6 gene transcription in IL-6 treatment conditions, which is further confirmed by ERRE mutated mBMP6-luciferase reporter assays. Finally, an inverse agonist of ERRγ, GSK5182, markedly inhibits IL-6 induced hepatic BMP6 expression in vitro and in vivo. Taken together, these results reveal a novel molecular mechanism on ERRγ mediated transcriptional regulation of hepatic BMP6 gene expression in response to IL-6.
骨形态发生蛋白 6(BMP6)是一种多功能生长因子,参与器官发育和稳态。BMP6 控制着肝脏激素铁调素的表达,因此在调节铁稳态方面起着至关重要的作用。BMP6 基因在肝脏中的转录调控很大程度上是未知的,但对于在病理条件下外部调节铁负荷将有很大帮助。在这里,我们描述了一种孤儿核受体雌激素相关受体γ(ERRγ)响应促炎细胞因子白细胞介素 6(IL-6)对肝脏 BMP6 基因表达的详细分子机制。重组 IL-6 处理增加了肝脏 ERRγ 和 BMP6 的表达。ERRγ 的过表达足以增加肝细胞中 BMP6 基因的表达,这表明 IL-6 是 ERRγ 的上游。与此一致,细胞系中 ERRγ 的敲低或小鼠肝脏特异性 ERRγ 的敲除显著降低了 IL-6 介导的 BMP6 表达。启动子研究表明,ERRγ 直接结合到小鼠 BMP6 基因启动子中的 ERR 反应元件(ERRE)上,并在 IL-6 处理条件下正向调节 BMP6 基因转录,这通过 ERRE 突变的 mBMP6-荧光素酶报告基因测定进一步得到证实。最后,ERRγ 的反向激动剂 GSK5182 显著抑制了体外和体内 IL-6 诱导的肝脏 BMP6 表达。总之,这些结果揭示了 ERRγ 介导的肝脏 BMP6 基因表达对 IL-6 反应的转录调控的新分子机制。