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

阐明非编码 RNA-LSD1 相互作用的结构和功能

• 批准号: 10393402
• 负责人: Nicholas J Reiter
• 金额: $ 0.85万
• 依托单位: MARQUETTE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10393402
• 关键词: Adopted; Aging; Architecture; Binding; Binding Proteins; Biochemical; Bladder; Cell Differentiation process; Cells; Chromatin; Chromosomal Stability; Chromosomes; DNA Double Strand Break; Degenerative Disorder; Development; Double Strand Break Repair; Enzymes; Epigenetic Process; Gene Expression; Gene Expression Regulation; Genes; Genome Stability; Genomic approach; Goals; Guanine; Heterochromatin; Human; In Vitro; KDM1A gene; Lead; Link; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Molecular; Neoplasm Metastasis; Neuroblastoma; Oncogenic; Pathway interactions; Play; Property; Prostate; Proteins; RNA; Regulator Genes; Regulatory Pathway; Repressor Proteins; Research Project Grants; Role; Structure; Telomere Shortening; Untranslated RNA; Yeasts; base; epigenetic regulation; histone methylation; human disease; in vivo; insight; malignant breast neoplasm; neuron development; repair enzyme; telomere; therapeutic development; therapeutic target

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.

摘要 该研究项目的核心假设是核糖核酸（RNA）结构和 RNA-染色质相关蛋白相互作用在维持基因组中起着重要作用 稳定非编码RNA（ncRNA）是基因表达的关键调节因子， 已知与染色质相关蛋白质物理结合以组织基因改变 在细胞的特定发育阶段的结构。这些RNA蛋白的一个子集 相互作用与衰老、癌症转移和神经元发育有关。如基因组 尽管已经有开始方法来鉴定RNA-蛋白质关联及其与癌症的联系， 有限的信息表观遗传为基础的RNA结合蛋白在分子水平上。我们 试图了解RNA如何在功能上与赖氨酸特异性脱甲基酶-1相互作用 （LSD 1）蛋白酶。LSD 1是一种重要的组蛋白甲基化调节剂，具有致癌作用。 在几种癌症中的特性，包括：前列腺癌、膀胱癌、神经母细胞瘤、肺癌和乳腺癌 癌LSD 1通过其庞大的相互作用网络与癌症有关。LSD 1还与 与许多RNA分子参与细胞分化，可以发挥调节保守的 细胞中RNA介导的阻遏物复合物。一种称为TERRA的ncRNA使LSD 1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LSD 1的RNA结合特性对于基因表达是重要的。 调控TERRA是一种调节RNA，从酵母转录到人类，可以通过 一个非典型的鸟嘌呤四链体（GQ）RNA结构在体外和体内。TERRA的 细胞中的丰度与端粒的进行性缩短和端粒的稳定性相关。 端粒异染色质我们假设RNA-LSD 1组装体在基因表达中起关键作用， 表达，并提出GQ RNA作为染色质相关的重要调控因子， proteins.拟议的研究将1）阐明生物化学和结构机制， TERRA RNA-LSD 1相互作用和2）检查非典型结构RNA如何接触 LSD 1影响端粒调控途径。该项目的长期目标是 了解RNA结构如何影响LSD 1调控网络。从这些 这些研究将深入了解基于RNA的表观遗传调控机制， 作为开发针对组蛋白甲基化的“定制”探针的起点， 以特定路径的方式进行监管。