The novel long noncoding RNA Lnc19959.2 modulates triglyceride metabolism -associated genes through the interaction with Purb and hnRNPA2B1
The novel long noncoding RNA Lnc19959.2 modulates triglyceride metabolism -associated genes through the interaction with Purb and hnRNPA2B1
复制标题
新型长非编码RNA Lnc19959.2通过与Purb和hnRNPA2B1的相互作用调节甘油三酯代谢相关基因
DOI：
10.1016/j.molmet.2020.100996
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发表时间：
2020-07-01
影响因子：
8.1
通讯作者：
Ding, Hu
中科院分区：
文献类型：
--
作者：
Wang, Jing;Xiang, Dao;Ding, Hu
目的长非编码RNA(Long Non Coding RNAs，LncRNAs)目前被认为具有重要而广泛的生物学功能，但甘油三酯代谢的分子机制尚不清楚。本研究旨在寻找在高甘油三酯血症大鼠肝脏中差异表达的新的lncRNAs，并阐明其在甘油三酯代谢中的功能作用。方法用转录组测序和实时荧光定量聚合酶链式反应(Real-time PCR)鉴定高甘油三酯血症大鼠肝脏差异表达的lncRNAs。通过体外和活体实验评价了nc19959.2在甘油三酯代谢中的作用，并通过RNA下拉和RIP实验评价了nc19959.2与其靶蛋白之间的相互作用。结果克隆到一个新的LncRNA，在高甘油三酯血症大鼠肝脏中表达上调。Nc19959.2基因敲除后，在体内具有明显的降甘油三酯作用。随后，全基因组分析证实，nc19959.2基因的敲除导致了许多基因在甘油三酯动态平衡期间的解除调控。进一步的机制研究表明，lnc19959.2通过泛素化的转录抑制因子Purb上调ApoA4的表达，而它与hnRNPA2B1特异性地相互作用，分别下调Cpt1a、Tm7sf2和Gpam的表达。在上游途径中，棕榈酸上调CCAAT/增强子结合蛋白Beta(CEBPB)，并促进其与nc19959.2启动子的结合，从而显著促进nc19959.2的转录活性。结论我们的研究结果为揭示lncRNA通过新的lncRNAlnc19959.2调控甘油三酯稳态的新层次调控复杂性提供了新的见解。
ObjectiveLong noncoding RNAs (lncRNAs) are currently considered to have a vital and wide range of biological functions, but the molecular mechanism underlying triglycerides metabolism remains poorly understood. This study aims to identify novel lncRNAs differentially expressed in rat livers with hypertriglyceridemia and elucidated the function role in TG metabolism.MethodsDifferentially expressions of lncRNAs in rat livers with hypertriglyceridemia were identified by transcriptome sequencing and validated by real-time PCR. The role oflnc19959.2in triglyceride metabolism was assessed bothin vitroandin vivo.RNA pulldown and RIP assays were conducted to evaluate the interactions betweenlnc19959.2and its target proteins. ChIP and Dual report assays were performed to detect the interactions between transcription factors and promoters of its target genes.ResultsWe identified a novel lncRNA, andlnc19959.2was upregulated in rat livers with hypertriglyceridemia. The knockdown oflnc19959.2 has profound TG lowering effectsin vitroandin vivo. Subsequently, the genome-wide analysis identified that the knockdown oflnc19959.2caused the deregulation of many genes during TG homeostasis. Further mechanism studies revealed thatlnc19959.2upregulatedApoA4expression via ubiquitinated transcription inhibitor factorPurb, while it specifically interacted withhnRNPA2B1to downregulate the expression ofCpt1a,Tm7sf2, andGpam, respectively. In the upstream pathway, palmitate acid upregulated CCAAT/Enhancer-Binding Protein Beta (Cebpb) and facilitated its binding to the promoter oflnc19959.2, which resulted in significant promotion oflnc19959.2transcriptional activity.ConclusionsOur findings provide novel insights into a new layer regulatory complexity of an lncRNA modulating triglyceride homeostasis by a novel lncRNAlnc19959.2.