Wang Zhongyan, Burke Peter A
Department of Surgery, Boston University School of Medicine, Boston, MA 02118, USA.
J Mol Biol. 2007 Aug 10;371(2):323-35. doi: 10.1016/j.jmb.2007.05.049. Epub 2007 May 24.
Following injury, a large number of hepatic acute phase genes are rapidly modulated at the transcriptional level to restore metabolic homeostasis and limit tissue damage. Hepatocyte nuclear factor 4alpha (HNF-4alpha) is a liver-enriched transcription factor that controls embryonic liver development and regulates tissue-specific gene expression in adult liver cells. Many genes encoding acute phase proteins contain HNF-4alpha-binding sites in their promoter regions and are transcriptionally regulated by HNF-4alpha. Utilizing a cytokine induced acute phase response in HepG2 cells, we investigated the role of HNF-4alpha in regulating the transcription of three HNF-4alpha sensitive genes, alpha1-antitrypsin (alpha1-AT), transthyretin (TTR), and apolipoprotein B (ApoB) after injury. The transcriptional behavior of all three genes depends, in part, on the intracellular concentrations of HNF-4alpha. However, the unique mRNA expression patterns of alpha1-AT, TTR, and ApoB in response to cytokine treatment were abrogated in HepG2 cells with dramatically reduced HNF-4alpha protein concentrations. The mechanism by which HNF-4alpha mediates this injury response is through site-specific alterations in HNF-4alpha-binding abilities and transactivation potentials. Cytokine treatment phosphorylates HNF-4alpha, which directly affects HNF-4alpha activity. Our results demonstrate that HNF-4alpha is a crucial mediator in the regulation of alpha1-AT, TTR, and ApoB gene expression before and after injury, providing evidence of a novel role for HNF-4alpha in the control of the liver's acute phase response.
损伤后,大量肝脏急性期基因在转录水平迅速被调节,以恢复代谢稳态并限制组织损伤。肝细胞核因子4α(HNF-4α)是一种肝脏富集的转录因子,它控制胚胎肝脏发育并调节成年肝细胞中组织特异性基因的表达。许多编码急性期蛋白的基因在其启动子区域含有HNF-4α结合位点,并受HNF-4α转录调控。利用HepG2细胞中细胞因子诱导的急性期反应,我们研究了HNF-4α在损伤后调节三个HNF-4α敏感基因,即α1-抗胰蛋白酶(α1-AT)、转甲状腺素蛋白(TTR)和载脂蛋白B(ApoB)转录中的作用。这三个基因的转录行为部分取决于HNF-4α的细胞内浓度。然而,在HNF-4α蛋白浓度显著降低的HepG2细胞中,α1-AT、TTR和ApoB对细胞因子治疗的独特mRNA表达模式被消除。HNF-4α介导这种损伤反应的机制是通过HNF-4α结合能力和反式激活潜能的位点特异性改变。细胞因子治疗使HNF-4α磷酸化,这直接影响HNF-4α的活性。我们的结果表明,HNF-4α是损伤前后α1-AT、TTR和ApoB基因表达调控中的关键介质,为HNF-4α在肝脏急性期反应控制中的新作用提供了证据。