Biophysics of regulatory RNAs and RNPs - Equipment Supplement

项目摘要 非编码RNA是真核生物多种生物学过程的关键调控因子。MicroRNAs是一类 小的非编码RNA，在转录后以序列依赖的方式调节基因的表达。 大约1,000个人类microRNA似乎控制着超过一半的人类 信使RNA。偏离动态平衡的microRNA表达水平，降低或增强 表达，与癌症、糖尿病、心脏病和神经退行性疾病等有关。至 要保持适当的microRNA表达水平，真核细胞必须严格控制 初级和前体microRNA元件。然而，这一严格规定背后的分子决定因素 对于microRNA的生物发生还没有完全了解。在本提案中，我们将探讨cis(RNA结构)和 MicroRNAs生物发生的反式(蛋白质结合伙伴)调节因子。在研究领域1，我们将探索 Let-7微RNA家族成员的转录后调控。处理的大约是 一半的let-7 microRNAs是由蛋白质伙伴Lin28介导的。这个家庭的其他成员是 受其他因素调节的，通常是未知的。我们将确定RNA修饰的程度， 结构，而RNA动态可以作为调控触发器来控制这些其他家族的加工 会员。在研究区域2中，我们将研究oncomiR-1，一种多顺反子初级生物的差异处理 在许多癌症中富含的微核糖核酸。我们将在细胞内进行化学探测研究，确定相关的 细胞蛋白质，并阐明控制亚区处理的结构开关。我们的长期目标 目的是单独确定oncomiR-1的三级结构，以及与调控蛋白形成复合体。总而言之， 这些研究将有助于阐明高度调控的生产背后的分子决定因素 MicroRNA。相对于蛋白质，RNA结构仍然确定不足，导致我们的 对RNA折叠和功能的机械理解。我们期望该方法和方法 这些研究的发展将广泛适用于其他调控RNA和蛋白质-RNA的研究 复合体。这种增强的结构洞察力将有助于了解RNA结构如何直接调节生物 并将为新的RNA靶向疗法的开发铺平道路。

Project Summary Non-coding RNAs are key regulators of diverse biological processes in eukaryotes. MicroRNAs are a family of small non-coding RNAs that post-transcriptionally regulate gene expression in a sequence-dependent manner. Approximately 1,000 human microRNAs appear to control the expression of more than half of all human messenger RNAs. Deviations from homeostatic microRNA expression levels, either reduced or enhanced expression, have been linked to cancers, diabetes, heart and neurodegenerative diseases, among others. To maintain proper microRNA expression levels, eukaryotic cells must tightly control the enzymatic processing of primary and precursor microRNA elements. However, the molecular determinants underlying this strict regulation of microRNA biogenesis are not fully understood. In this proposal we will explore both cis (RNA structure) and trans (protein binding partners) regulators of microRNAs biogenesis. In Research Area 1, we will explore the post-transcriptional regulation of members of the let-7 family of microRNAs. The processing of approximately half of the let-7 microRNAs is mediated by a protein partner, Lin28. The other members of this family are regulated by other, often unknown factors. We will determine the extent to which RNA modification, RNA structure, and RNA dynamics can serve as regulatory triggers to control the processing of these other family members. In Research Area 2, we will examine the differential processing of oncomiR-1, a polycistronic primary microRNA that is enriched in many cancers. We will conduct in cell chemical probing studies, identify associated cellular proteins, and elucidate a structural switch that controls processing of a subdomain. Our long-term goal is to determine the tertiary structure of oncomiR-1 alone and in complex with regulatory proteins. Collectively, these studies will help elucidate the molecular determinants underlying the highly-regulated production of microRNAs. RNA structures remain underdetermined relative to proteins, leading to an asymmetry in our mechanistic understanding of RNA folding and function. We expect that the methodology and approach developed in these studies will be broadly applicable to studies of other regulatory RNAs and protein-RNA complexes. This enhanced structural insight will inform on how RNA structure can directly regulate biological activity and will pave the way for the development of novel RNA-targeted therapeutics.