Elucidating the Structure and the Function of Non-Coding RNA-LSD1 Interactions

摘要 这项研究的中心假设是核糖核酸(RNA)的结构和 RNA-染色质相关蛋白相互作用在维持基因组中起着至关重要的作用 稳定性。非编码RNA(NcRNAs)是基因表达的关键调节因子， 已知与染色质相关蛋白物理结合以组织基因变化 架构在细胞的特定发育阶段。这些RNA-蛋白质的一个子集 相互作用与衰老、癌症转移和神经元发育有关。作为基因组 方法开始确定RNA-蛋白质的关联及其与癌症的联系，仍然存在 关于基于表观遗传的RNA结合蛋白在分子水平上的有限信息。我们 试图了解RNA如何在功能上与赖氨酸特异性去甲基酶-1相互作用 (LSD1)蛋白酶。LSD1是一种重要的组蛋白甲基化调节因子，具有致癌作用 几种癌症的特性，包括：前列腺癌、膀胱癌、神经母细胞瘤、肺癌和乳腺癌 癌症。LSD1通过其庞大的相互作用网络与癌症有关。LSD1还可以交互 有许多RNA分子参与细胞分化，并能调节保守的 细胞内的RNA介导的阻遏复合体。一种称为Terra的ncRNA使LSD1能够 与染色体末端的DNA双链断裂修复酶相互作用 (端粒)，表明LSD1的RNA结合特性对基因是重要的 监管。Terra是一种从酵母转录到人类的调节性RNA，可以采用 体外和体内非规范的鸟嘌呤四链(GQ)RNA结构。Terra‘s 细胞中的丰度与进行性端粒缩短和端粒稳定有关 端粒异染色质。我们假设RNA-LSD1组装在基因中起关键作用 表达并提出GQ RNAs是染色质相关的重要调节因子 蛋白质。拟议的研究将1)阐明一种生物化学和结构机制 Terra RNA-LSD1相互作用和2)检查非规范结构的RNA如何接触 LSD1影响端粒调控通路。这个项目的长期目标是 了解RNA结构如何影响LSD1调节网络。来自这些的结果 研究将提供对基于RNA的表观遗传调控和服务的机制的洞察 作为开发以组蛋白甲基化为靶点的“量身定制”探针的起点 以一种特定途径的方式调节。

Abstract This research project is centered on the hypothesis that ribonucleic acid (RNA) structure and RNA-chromatin associated protein interactions play crucial roles in maintaining genome stability. Non-coding RNAs (ncRNAs) serve as key regulators of gene expression and are known to physically associate with chromatin-associated proteins to organize changes in gene architecture during specific development stages of the cell. A subset of these RNA-protein interactions are linked to ageing, cancer metastasis, and neuronal development. As genomic approaches begin to identify RNA-protein associations and their links to cancer, there remains limited information about epigenetic-based RNA binding proteins at the molecular level. We seek to understand how RNA functionally interacts with the lysine specific demethylase-1 (LSD1) protein enzyme. LSD1 is an essential histone methylation regulator and has oncogenic properties in several cancers including: prostate, bladder, neuroblastoma, lung, and breast cancer. LSD1 is implicated in cancer through its vast interaction network. LSD1 also interacts with many RNA molecules involved in cell differentiation and can function to regulate conserved RNA-mediated repressor complexes in the cell. An ncRNA, termed TERRA, enables LSD1 to interact with a DNA double strand break repair enzyme at the ends of chromosomes (telomeres), demonstrating that the RNA binding properties of LSD1 are important for gene regulation. TERRA is a regulatory RNA that is transcribed from yeast to humans and can adopt a non-canonical guanine quadruplex (GQ) RNA architecture in vitro and in vivo. TERRA’s abundance in the cell correlates with progressive telomere shortening and the stabilization of telomeric heterochromatin. We hypothesize that RNA-LSD1 assemblies play key roles in gene expression and propose that GQ RNAs serve as crucial regulators of chromatin-associated proteins. The proposed studies will 1) elucidate the biochemical and structural mechanisms of a TERRA RNA-LSD1 interaction and 2) examine how non-canonical structured RNAs contact LSD1 to influence telomeric regulatory pathways. The long-term goal of this project is to understand how RNA structure influences LSD1 regulatory networks. Results from these studies will provide insight into the mechanisms of RNA-based epigenetic regulation and serves as a starting point in the development of ‘tailored’ probes that target histone methylation regulators in a pathway-specific manner.