Characterization of the mammalian mRNA 3'-end processing complex

项目总结： 这项建议的长期目标是详细了解 哺乳动物mRNA3‘的加工及其调控。信使核糖核酸3‘端形成，通常 涉及内切核裂解和随后的多聚腺苷，是必不可少的一步。 以及它对RNA的许多方面都有显著影响 代谢，包括信使核糖核酸的稳定性、亚细胞定位和翻译。在……里面 此外，大多数真核基因产生多种不同的mrna异构体。 3‘末端通过交替多聚腺苷(APA)。最近的研究表明， APA在发育过程中受到高度调控，并在后发育中发挥重要作用 转录基因调控。异常的APA模式与广泛的 一系列疾病，从癌症到神经紊乱。两个突出的关键问题 在mRNA3‘加工领域的问题是：1)什么是分子 信使核糖核酸3‘加工机制(包括聚(A)位(PAS)的加工机制) 裂解和多聚腺苷的识别和催化)？2)PAS如何？ 选择还是受APA监管？要解决这些根本问题，至关重要的是 了解mRNA3‘加工因子的结构与功能关系。在……里面 在前几个资助期发表的研究报告中，我们重新编制了关键 哺乳动物mRNA 3‘处理复合体的模块，表征了AAUAAA如何 和富U/Gu的下游元素(这两个关键元素定义了 哺乳动物PAS)被识别，并揭示了mRNA3‘加工和剪接 可以通过类似的机制进行调节。基于这些发现，我们在这里 建议定义完全重构的人类的结构-功能关系 MRNA3‘处理复合体并系统地表征RNA结合的作用 调节Pas选择和APA的蛋白质(RBPs)。 好了！

Project summary: The long-term goal of this proposal is to understand, in detail, the mechanisms of mammalian mRNA 3' processing and its regulation. mRNA 3'-end formation, typically involving an endonucleolytic cleavage followed by polyadenylation, is an essential step of eukaryotic gene expression and it significantly impacts many aspects of RNA metabolism, including mRNA stability, subcellular localization and translation. In addition, the majority of eukaryotic genes produce multiple mRNA isoforms with distinct 3' ends through alternative polyadenylation (APA). Recent studies have revealed that APA is highly regulated in development and plays an important role in post- transcriptional gene regulation. Aberrant APA patterns have been associated with a wide range of diseases, from cancer to neurological disorders. Two key outstanding questions in the mRNA 3' processing field have been: 1) what is the molecular mechanism of mRNA 3' processing (including mechanisms for poly(A) site (PAS) recognition and catalysis of cleavage and polyadenylation)? 2) how is PAS selection or APA regulated? To address these fundamental questions, it is essential to understand the structure-function relationship of mRNA 3' processing factors. In published studies during the previous funding periods, we have reconstituted key modules of the mammalian mRNA 3' processing complex, characterized how AAUAAA and the U/GU-rich downstream element (two key elements that define the majority of mammalian PAS) are recognized, and revealed that mRNA 3' processing and splicing can be regulated through a similar mechanism. Building on these findings, here we propose to define the structure-function relationship of the fully reconstituted human mRNA 3' processing complex and systematically characterize the role of RNA-binding proteins (RBPs) in regulating PAS selection and APA. !