National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of the Chinese Academy of Sciences, Beijing 100049, China.
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
Biochim Biophys Acta Gene Regul Mech. 2019 Feb;1862(2):209-218. doi: 10.1016/j.bbagrm.2018.11.006. Epub 2018 Nov 30.
As an RNA binding protein, CUG-BP Elav-like family (CELF) has been shown to be critical for heart biological functions. However, no reports have revealed the function of CELF1 in hypertrophic cardiomyopathy (HCM). Hinted by RNA immunoprecipitation-sequencing (RIP-seq) data, the influence of the CELF protein on heme oxygenase-1 (HO-1) expression was tested by modulating CELF1 levels. Cardiac hypertrophy is related to oxidative stress-induced damage. Hence, the cardiovascular system may be protected against further injury by upregulating the expression of antioxidant enzymes, such as HO-1. During the past two decades, research has demonstrated the central role of HO-1 in the protection against diseases. Thus, understanding the molecular mechanisms underlying the modulation of HO-1 expression is profoundly important for developing new strategies to prevent cardiac hypertrophy. To elucidate the molecular mechanisms underlying HO-1 regulation by the CELF protein, we performed RNA immunoprecipitation (RIP), biotin pull-down analysis, luciferase reporter and mRNA stability assays. We found that the expression of HO-1 was downregulated by CELF1 through the conserved GU-rich elements (GREs) in HO-1 3'UTR transcripts. Correspondingly, CELF1 expression was regulated by controlling the release of carbon monoxide (CO) in H9C2 cells. The CELF1-HO-1-CO regulation axis constituted a novel positive feedback circuit. In addition, we detected the potential involvement of CELF1 and HO-1 in samples from HCM patients. We found that CELF1 and CELF2, but not HO-1, were highly expressed in HCM heart samples. Thus, a manipulation targeting CELF1 could be developed as a potential therapeutic option for cardiac hypertrophy.
作为一种 RNA 结合蛋白,CUG-BP Elav 样家族(CELF)对于心脏的生物学功能至关重要。然而,目前尚无报道揭示 CELF1 在肥厚型心肌病(HCM)中的功能。RNA 免疫沉淀测序(RIP-seq)数据提示,通过调节 CELF1 水平来测试 CELF 蛋白对血红素加氧酶-1(HO-1)表达的影响。心脏肥大与氧化应激诱导的损伤有关。因此,通过上调抗氧化酶(如 HO-1)的表达,心血管系统可能会免受进一步损伤。在过去的二十年中,研究表明 HO-1 在保护疾病方面发挥着核心作用。因此,了解 HO-1 表达调控的分子机制对于开发预防心脏肥大的新策略具有重要意义。为了阐明 CELF 蛋白调节 HO-1 表达的分子机制,我们进行了 RNA 免疫沉淀(RIP)、生物素下拉分析、荧光素酶报告基因和 mRNA 稳定性测定。我们发现,HO-1 的表达通过 HO-1 3'UTR 转录本中的保守 GU 丰富元件(GREs)被 CELF1 下调。相应地,CELF1 的表达通过控制 H9C2 细胞中一氧化碳(CO)的释放来调节。CELF1-HO-1-CO 调节轴构成了一个新的正反馈回路。此外,我们在 HCM 患者的样本中检测到了 CELF1 和 HO-1 的潜在参与。我们发现 CELF1 和 CELF2,但不是 HO-1,在 HCM 心脏样本中高表达。因此,针对 CELF1 的操作可以作为心脏肥大的潜在治疗选择。