Department of Pharmacology, College of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee 38163.
Department of Pharmacology, College of Medicine, University of Tennessee Health Science Center, Memphis, Tennessee 38163; Department of Veterans Affairs Medical Center, Memphis, Tennessee 38163.
J Biol Chem. 2013 Jun 7;288(23):16321-16333. doi: 10.1074/jbc.M112.445569. Epub 2013 Apr 29.
Secretory phospholipase A2 group IIa (PLA2g2a) is associated with inflammation, hyperlipidemia, and atherogenesis. Transcription of the PLA2g2a gene is induced by multiple cytokines. Here, we report the surprising observation that thyroid hormone (T3) inhibited PLA2g2a gene expression in human and rat hepatocytes as well as in rat liver. Moreover, T3 reduced the cytokine-mediated induction of PLA2g2a, suggesting that the thyroid status may modulate aspects of the inflammatory response. In an effort to dissect the mechanism of repression by T3, we cloned the PLA2g2a gene and identified a negative T3 response element in the promoter. This T3 receptor (TRβ)-binding site differed considerably from consensus T3 stimulatory elements. Using in vitro and in vivo binding assays, we found that TRβ bound directly to the PLA2g2a promoter as a heterodimer with the retinoid X receptor. Knockdown of nuclear corepressor or silencing mediator for retinoid and thyroid receptors by siRNA blocked the T3 inhibition of PLA2g2a. Using chromatin immunoprecipitation assays, we showed that nuclear corepressor and silencing mediator for retinoid and thyroid receptors were associated with the PLA2g2a gene in the presence of T3. In contrast with the established role of T3 to promote coactivator association with TRβ, our experiments demonstrate a novel inverse recruitment mechanism in which liganded TRβ recruits corepressors to inhibit PLA2g2a expression.
分泌型磷脂酶 A2 组 IIa(PLA2g2a)与炎症、高血脂和动脉粥样硬化形成有关。PLA2g2a 基因的转录受多种细胞因子诱导。在这里,我们报告了一个令人惊讶的观察结果,即甲状腺激素(T3)抑制了人源和鼠源肝细胞以及鼠肝中 PLA2g2a 基因的表达。此外,T3 降低了细胞因子介导的 PLA2g2a 诱导,这表明甲状腺状态可能调节炎症反应的某些方面。为了剖析 T3 抑制的机制,我们克隆了 PLA2g2a 基因,并在启动子中鉴定了一个负性 T3 反应元件。这个 T3 受体(TRβ)结合位点与共识 T3 刺激元件有很大的不同。通过体外和体内结合实验,我们发现 TRβ 作为异二聚体与视黄酸 X 受体直接结合到 PLA2g2a 启动子上。通过 siRNA 敲低核共抑制因子或视黄酸和甲状腺受体沉默调节剂可阻断 T3 对 PLA2g2a 的抑制作用。通过染色质免疫沉淀实验,我们表明在 T3 存在的情况下,核共抑制因子和视黄酸和甲状腺受体沉默调节剂与 PLA2g2a 基因相关。与 T3 促进共激活因子与 TRβ 结合的既定作用相反,我们的实验证明了一种新的反向招募机制,即配体结合的 TRβ 招募共抑制因子来抑制 PLA2g2a 的表达。