Garcia Manon, Holota Hélène, De Haze Angélique, Saru Jean-Paul, Sanchez Phelipe, Battistelli Edwige, Thirouard Laura, Monrose Mélusine, Benoit Gérard, Volle David H, Beaudoin Claude
Université Clermont Auvergne, CNRS UMR6293, INSERM U1103, iGReD Team-Volle, Clermont-Ferrand, France.
Université de Rennes 1, CNRS UMR6290, INSERM U1305, IGDR, Rennes Cedex, France.
J Biol Chem. 2025 Jan;301(1):108022. doi: 10.1016/j.jbc.2024.108022. Epub 2024 Nov 26.
Farnesoid X receptor α (FXRα, NR1H4) is a bile acid-activated nuclear receptor that regulates the expression of glycolytic and lipogenic target genes by interacting with the 9-cis-retinoic acid receptor α (RXRα, NR2B1). Along with cofactors, the FXRα proteins reported thus far in humans and rodents have been observed to regulate both isoform (α1-4)- and tissue-specific gene expression profiles to integrate energy balance and metabolism. Here, we studied the biological functions of an FXRα naturally occurring spliced exon 5 isoform (FXRαse5) lacking the second zinc-binding module of the DNA-binding domain. We demonstrate spliced exon 5 FXRα expression in all FXRα-expressing human and mouse tissues and cells, and that it is unable to bind to its response element or activate FXRα dependent transcription. In parallel, this spliced variant displays differential interaction capacities with its obligate heterodimer partner retinoid X receptor α that may account for silencing of this permissive dimer for signal transduction. Finally, deletion of exon 5 by gene edition in HepG2 cells leads to FXRα loss-of-function, increased expression of LRH1 metabolic sensor and CD36 fatty acid transporter in conjunction with changes in glucose and triglycerides homeostasis. Together, these findings highlight a novel mechanism by which alternative splicing may regulate FXRα gene function to fine-tune adaptive and/or metabolic responses. This finding deepens our understanding on the role of splicing events in hindering FXRα activity to regulate specific transcriptional programs and their contribution in modifying energy metabolism in normal tissues and metabolic diseases.
法尼酯X受体α(FXRα,NR1H4)是一种胆汁酸激活的核受体,它通过与9-顺式视黄酸受体α(RXRα,NR2B1)相互作用来调节糖酵解和脂肪生成靶基因的表达。与辅因子一起,迄今为止在人类和啮齿动物中报道的FXRα蛋白已被观察到可调节异构体(α1-4)和组织特异性基因表达谱,以整合能量平衡和代谢。在这里,我们研究了一种天然存在的剪接外显子5异构体(FXRαse5)的生物学功能,该异构体缺乏DNA结合结构域的第二个锌结合模块。我们证明了剪接外显子5 FXRα在所有表达FXRα的人类和小鼠组织及细胞中的表达,并且它无法结合其反应元件或激活FXRα依赖性转录。同时,这种剪接变体与其专一性异二聚体伙伴视黄酸X受体α表现出不同的相互作用能力,这可能解释了这种允许性二聚体对信号转导的沉默作用。最后,通过基因编辑在HepG2细胞中缺失外显子5会导致FXRα功能丧失,同时LRH1代谢传感器和CD36脂肪酸转运蛋白的表达增加,并伴有葡萄糖和甘油三酯稳态的变化。总之,这些发现突出了一种新机制,即可变剪接可能调节FXRα基因功能以微调适应性和/或代谢反应。这一发现加深了我们对剪接事件在阻碍FXRα活性以调节特定转录程序中的作用及其在正常组织和代谢疾病中改变能量代谢方面的贡献的理解。