Characterization of the mammalian mRNA 3'-end processing complex

哺乳动物 mRNA 3 端加工复合物的表征

• 批准号: 10580430
• 负责人: Yongsheng Shi
• 金额: $ 13.32万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-15 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10580430
• 关键词: Address; Catalysis; Complex; Development; Disease; Elements; Funding; Gene Expression; Gene Expression Regulation; Genes; Goals; Human; Malignant Neoplasms; Messenger RNA; Molecular; Pattern; Play; Poly A; Polyadenylation; Post-Transcriptional Regulation; Protein Isoforms; Publishing; RNA Splicing; RNA metabolism; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Role; Site; Structure-Activity Relationship; Translations; human disease; mRNA Stability; nervous system disorder; reconstitution

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.

项目概要： 本提案的长期目标是详细了解 哺乳动物mRNA的3'加工及其调控。mRNA 3 '-末端形成，通常 涉及核酸内切切割，然后是聚腺苷酸化，是一个重要步骤 真核基因表达的影响，它显着影响RNA的许多方面， 代谢，包括mRNA的稳定性，亚细胞定位和翻译。在 此外，大多数真核基因产生多种mRNA同种型， 3'末端通过交替聚腺苷酸化（阿帕）。最近的研究表明， 阿帕在发育过程中受到高度调控，在后发育过程中发挥着重要作用。 转录基因调控异常的阿帕模式与广泛的 从癌症到神经系统疾病。两个关键突出 mRNA 3'加工领域的问题是：1）什么是mRNA 3'加工的分子机制？ mRNA 3'加工机制（包括多聚腺苷酸位点（PAS）机制） 切割和多聚腺苷酸化的识别和催化）？2)PAS怎么样 选择还是阿帕监管？为了解决这些基本问题，必须 了解mRNA 3'加工因子的结构与功能关系。在 在过去的资助期间发表的研究，我们已经重建了关键 哺乳动物mRNA 3'加工复合物的模块，其特征在于AAUAAA如何 和富含铀/铀的下游元素（两个关键元素，定义了大多数 哺乳动物PAS）的识别，并揭示mRNA 3'加工和剪接， 也可以通过类似的机制进行调节。基于这些发现，我们 建议定义完全重组人体的结构-功能关系 mRNA 3'加工复合物，并系统地表征RNA结合的作用 RBP在调节PAS选择和阿帕中的作用。

项目成果

The end of the message: multiple protein-RNA interactions define the mRNA polyadenylation site.

• DOI: 10.1101/gad.261974.115
• 发表时间: 2015-05-01
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Shi Y;Manley JL
• 通讯作者: Manley JL

CPSF30 and Wdr33 directly bind to AAUAAA in mammalian mRNA 3' processing.

• DOI: 10.1101/gad.250993.114
• 发表时间: 2014-11-01
• 期刊: Genes & development
• 影响因子: 10.5
• 作者: Chan SL;Huppertz I;Yao C;Weng L;Moresco JJ;Yates JR 3rd;Ule J;Manley JL;Shi Y
• 通讯作者: Shi Y

Crosstalk Between mRNA 3'-End Processing and Epigenetics.

• DOI: 10.3389/fgene.2021.637705
• 发表时间: 2021
• 期刊: Frontiers in genetics
• 影响因子: 3.7
• 作者: Soles LV;Shi Y
• 通讯作者: Shi Y

The TCF C-clamp DNA binding domain expands the Wnt transcriptome via alternative target recognition.

• DOI: 10.1093/nar/gku1186
• 发表时间: 2014-12-16
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Hoverter NP;Zeller MD;McQuade MM;Garibaldi A;Busch A;Selwan EM;Hertel KJ;Baldi P;Waterman ML
• 通讯作者: Waterman ML

Nudt21 Controls Cell Fate by Connecting Alternative Polyadenylation to Chromatin Signaling.

• DOI: 10.1016/j.cell.2017.11.023
• 发表时间: 2018-01-11
• 期刊: Cell
• 影响因子: 64.5
• 作者: Brumbaugh J;Di Stefano B;Wang X;Borkent M;Forouzmand E;Clowers KJ;Ji F;Schwarz BA;Kalocsay M;Elledge SJ;Chen Y;Sadreyev RI;Gygi SP;Hu G;Shi Y;Hochedlinger K
• 通讯作者: Hochedlinger K