Novel RNA-mediated regulatory roles of the human hnRNP L proteins

人类 hnRNP L 蛋白的新 RNA 介导的调节作用

• 批准号: 80033308
• 负责人: Professor Dr. Albrecht Bindereif
• 金额: --
• 依托单位: Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2008
• 资助国家: 德国
• 起止时间: 2007-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/80033308?language=en
• 关键词: Novel; RNA; mediated; regulatory; roles

The heterogenous nuclear ribonucleoprotein (hnRNP) L acts as a multifunctional RNA-binding protein with four RNA-recognition domains (RRMs), recognizing CA-rich RNA elements and activating or repressing alternative splicing in a position-dependent manner. As we have shown for CD45 alternative splicing in human B-cells, hnRNP L sometimes collaborates with hnRNP-like, a closely related paralog with the same domain organization. In addition, hnRNP L can regulate RNA stability, and our recent transcriptome-wide iCLIP analysis revealed global widespread binding in 3'-untranslated regions (3'UTRs) of mRNAs. On this basis we plan to characterize novel RNA-mediated roles of the hnRNP L proteins: First, we will address the potential role of hnRNP L in binding within 3'UTRs of mRNAs, focussing on competitive hnRNP L/microRNA binding, in particular its relevance for mRNA stability and translation. Second, we will extend our iCLIP approach to the hnRNP L-like paralog, to comprehensively analyze the hnRNP L/L-like cooperation in human B-cells. These genome-wide in vivo binding data will be complemented by an in vitro SELEX-Seq approach, focussing on the differential RNA-binding activities of the hnRNP L proteins and the combinatorial RNA recognition of their four RRMs. Third, we will explore a novel, potential function of the hnRNP L paralogs, binding to circular RNA sponges; we plan both to search for natural examples and to design artificial, hnRNP L/LL-specific circRNA sponges.

异源核核糖核蛋白L是一种多功能核糖核蛋白，具有4个核糖核酸识别结构域，识别富含CA的核糖核蛋白元件，以位置依赖的方式激活或抑制选择性剪接。正如我们已经在人类B细胞中显示的CD45选择性剪接，hnRNP L有时与hnRNP样合作，这是与相同结构域组织密切相关的平行序列。此外，hnRNP L还可以调节RNA的稳定性，我们最近在转录组范围内的iCLIP分析揭示了mRNAs的3‘非翻译区(3’UTRs)在全球范围内广泛结合。在此基础上，我们计划研究hnRNP L蛋白在RNA介导的新功能中的作用：首先，我们将讨论hnRNP L在mRNA3‘UTRs结合中的潜在作用，重点是竞争hnRNP L/microRNA结合，特别是它与mRNA稳定性和翻译的相关性。其次，我们将iCLIP方法扩展到hnRNP L样并行，全面分析hnRNP L/L样人类B细胞中的合作。这些全基因组的体内结合数据将由体外SELEX-SEQ方法补充，重点是hnRNP L蛋白的差异RNA结合活性及其四个RRMS的组合RNA识别。第三，我们将探索hnRNP L对偶的一个新的潜在功能，与环状RNA海绵结合；我们计划搜索自然例子和设计人工的，hnRNP L/LL特异的CircRNA海绵。