Trans-Regulation of RNA-Binding Protein Motifs by MicroRNA

 描述(申请人提供)：我们已经发现microoRNAs可以动态地调节RNA结合蛋白(RBP)结合位点，并表明miRNAs间接或直接以动态方式调节RBP结合位点，增加了转录后调控密码的复杂性和层次性。这个模型，我们称之为“sxRNA”的结构相互作用的RNA，代表了一个令人兴奋的新的转录后调控范式，可能提供基因表达调控的基本方面。我们已经从信息上确定了几个“反式作用”的sxRNA，我们的初步数据表明，积极和消极的变化都是可能的。负的或“抑制”的sxRNA复合体的例子抑制RBP结合活性，而正的或“诱导型”版本的sxRNA复合体可能增加RBP结合，并可使用RBP免疫沉淀(RIP)和miRNA分析来捕获。这种新的调控机制可能解释了包括选择性剪接在内的基因多功能的复杂调控是如何在转录后水平上被调控的。此外，这些相互作用的组合可以以模块化和组合的方式使用，从而增加了转录后调控密码的层级复杂性。与令人印象深刻的microRNA领域类似，sxRNA有可能增加我们对基因调控的理解，同时创造技术机会，将这种调控用作组织特异性mRNA，以替代基因治疗、抗病毒、基于RNA的分子工具、诊断甚至治疗，利用这些新发现的转录后方法。该提案的目标是测试一种新的转录后模型，在该模型中，miRNA和限制性商业惯例通过竞争反式来影响RNA结构。该项目旨在通过实验和信息学方法识别数十(可能数百)自然发生的microRNA-mRNA-RBP相互作用，表征sxRNA反式RNA相互作用的结构性质及其对RBP结合的影响，并使用专门设计的MS2-sxRNA报告程序验证它们在细胞系统中的存在和在翻译中的作用。

 DESCRIPTION (provided by applicant): We have discovered that micoRNAs can modulate RNA-binding protein (RBP) binding sites in a dynamic manner and suggest that miRNAs indirectly or directly modulate RBP-binding sites in a dynamic manner adding complexity and hierarchy to a post-transcriptional regulatory code. This model, which we call "sxRNA" for structurally interacting RNA, represents an exciting new post-transcriptional regulatory paradigm that may provide fundamental aspects of gene expression regulation. We have informatically identified several "trans-acting" sxRNAs and our preliminary data suggests that both positive and negative variations are possible. Examples of negative or "repressor" sxRNA complexes inhibit RBP binding activity, while positive or "inducer" versions of sxRNA complexes may increase RBP binding and can be captured using RBP immunoprecipatation (RIP) followed by miRNA analysis. This novel regulatory mechanism may explain how complex regulation of gene multi-functionality including alternative splicing could be regulated at the post-transcriptional level. Furthermore, combinations of these interactions could be used in a modular and combinatorial manner adding a hierarchical complexity to the post-transcriptional regulatory code. Similar to the impressive microRNA field, sxRNA has the potential to increase our understanding of gene regulation while simultaneously creating technological opportunities to utilize this regulation as a tissue specific mRNA alternative to gene therapy, anti-viral, RNA-based molecular tool, diagnostic, or even therapeutic that exploits these newly discovered post-transcriptional approaches. The goal of this proposal is to test a new post-transcriptional model in which miRNA and RBPs compete in trans to influence RNA structure. This project is designed to identify dozens (possibly hundreds) of naturally occurring microRNA-mRNA-RBP interactions by exploiting both experimental and informatic approaches, characterize the structural nature of the sxRNA trans-RNA interaction and its effect on RBP binding and validate their existence and role in translation in a cellular system using a specifically designed MS2-sxRNA reporter.