Exploring functional complexes and disease networks within human RNA-binding protein interactomes

项目总结 RNA结合蛋白(RBP)与编码和非编码RNA结合，影响RNA生命的每一步- 循环，包括前mRNA的加工，RNA的定位，以及翻译和降解的控制。更多和 更多的证据表明，RNA代谢障碍是许多人类神经退行性变的标志 包括肌萎缩侧索硬化症在内的疾病。了解RBP如何在协调网络中发挥作用 调控RNA的命运是揭示这些疾病病理基础的分子机制的关键。一个 人类限制性商业惯例的完整目录是难以捉摸的，因为出现了与 非多聚腺苷化的前mRNAs或缺乏特征的RNA结合域的非规范限制性商业惯例，因此 避开传统的RNA-相互作用组捕获研究。我们开发了一个计算型RBP分类器，它基于 观察到依赖于RNA分子共结合的蛋白质-蛋白质相互作用网络可以是 用于发现新的限制性商业惯例类别。这项提议寻求利用来自该分类器的预测来 完成三个主要目标。 1.阶段1：确定与疾病相关的非典型性RNA结合的生物学功能 核质转运相关候选基因RBP RANGAP1。 2.第二阶段：确定RNA结合对扩大的疾病组的生物学功能- 相关的非正则核质转运相关候选限制性商业惯例。 3.第二阶段：构建以RBP为中心的蛋白质-蛋白质相互作用网络，以扩大人类的蛋白质库 RBPS用于表征ALS相关的高阶RNP复合体。 我在DNA和RNA结合蛋白研究方面的丰富经验使我成为 执行此处建议的研究。这些目标将有助于完成一份全面的 人类限制性商业惯例，将有助于提供RNA调控网络的详细地图。加州大学圣迭戈分校的Yeo实验室是 在RNA生物学领域处于领先地位，因此是进行拟议培训的极佳环境 并建立一个独立的研究计划。Yeo实验室位于一家主要的生物医学中心 加州大学圣迭戈分校的研究中心，毗邻索尔克研究所，以及其他研究机构和生物技术 拉荷亚的公司。在这里实施拟议的培训计划将使我能够接触到STEM中的领导者 细胞生物学和蛋白质组学方法，我希望在向独立过渡的过程中掌握这些方法。

PROJECT SUMMARY RNA binding proteins (RBPs) bind to both coding and non-coding RNA to influence every step of the RNA life- cycle, including pre-mRNA processing, RNA localization, and control of translation and degradation. More and more evidence reveals that disruption of RNA metabolism is a hallmark of many human neurodegenerative diseases including Amyotrophic Lateral Sclerosis. Understanding how RBPs act in coordinated networks to regulate RNA fate is key to uncovering the molecular mechanisms underlying these disease pathologies. A complete catalogue of human RBPs is elusive due to the emergence of new classes of RBPs that interact with unpolyadenylated pre-mRNAs or non-canonical RBPs that lack characterized RNA-binding domains, and thus evade traditional RNA-interactome capture studies. We developed a computational RBP classifier based on the observation that protein-protein interaction networks that depend on co-binding of RNA molecules can be used to discover new classes of RBPs. This proposal seeks to utilize predictions from this classifier to accomplish 3 main goals. 1. Phase 1: Determine the biological function of RNA-binding by the disease-associated non-canonical nucleocytoplasmic transport related candidate RBP RANGAP1. 2. Phase 2: Determine the biological function of RNA-binding for an expanded group of disease- associated non-canonical nucleocytoplasmic transport related candidate RBPs. 3. Phase 2: Build an RBP centered protein-protein interaction network to expand the repertoire of human RBPs to characterize ALS relevant higher order RNP complexes. My extensive experience in the study of DNA and RNA binding proteins makes me an ideal candidate to perform the research proposed here. These aims will build towards the completion of a comprehensive list of human RBPs that will help provide detailed maps of RNA regulatory networks. The Yeo lab at UCSD is a leader in the field of RNA biology, and therefore is an excellent environment to perform the proposed training and build an independent research program. The Yeo lab is situated at the heart of a major biomedical research hub at UCSD, adjacent to the Salk Institute, and other research institutes and biotechnology companies in La Jolla. Conducting the proposed training program here will give me access to leaders in stem cell biology and proteomic methods that I hope to master as I transition to independence.