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Atomic-Resolution Analysis of eIF3-Mediated Translation Control

eIF3 介导的翻译控制的原子分辨率分析

基本信息

批准号：
10357567
负责人：
JAMIE H CATE
金额：
$ 36.17万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10357567
关键词：
5&apos Untranslated Regions Address Binding Binding Sites Biochemical Biochemistry Biological Cell Therapy Cell physiology Cells Chemicals Code Complex Couples Cryoelectron Microscopy Dependence Development Eukaryota Eukaryotic Initiation Factors Event Foundations Funding Future Gatekeeping Genes Genetic Transcription Genetic Translation Genotype Growth Human Human Biology In Vitro Individual Initiator Codon Maps Mass Spectrum Analysis Mediating Mediator of activation protein Messenger RNA Methods Modeling Modification Molecular Molecular Machines Mutate Nature Open Reading Frames Output Peptide Initiation Factors Phenotype Physiology Play Positioning Attribute Prokaryotic Initiation Factor-3 Protein Biosynthesis Proteins RNA Reader Reading Regulation Regulatory Pathway Reporter Repression Research Resolution Ribosomes Role Scanning Signal Transduction Solid Specificity Structure System Systems Biology Therapeutic Intervention Transcript Translating Translation Initiation Translations Untranslated Regions Writing base biological systems cell growth regulation cellular engineering crosslink dimethyl sulfate insight jun Oncogene protein expression recruit scaffold small molecule structural biology therapeutic target transcriptome

项目摘要

PROJECT SUMMARY Protein biosynthesis directly couples genotype to phenotype in the cell, and its regulation is central to cellular physiology. Our understanding of the molecular mechanism of protein synthesis has undergone a revolution in the last decade, built on rapid advances in structural biology and systems biology. However, important questions relating to dynamic events in translation remain unanswered, as the transient nature of these events makes them difficult to isolate. In this application, we propose to decipher the molecular mechanisms of how eukaryotic translation initiation factor 3 (eIF3) in humans regulates translation initiation. We recently discovered that human eIF3 serves multiple roles in translation. Human eIF3 generally helps assemble translation preinitiation complexes at the start codon, but also directly controls the translation of specific mRNAs. We found that eIF3 can either activate or repress the translation of an mRNA, depending on how eIF3 binds to the mRNA's 5' untranslated region (5' UTR). Furthermore, we discovered that eIF3 harbors a subunit–EIF3D–that binds the 5'-m7G cap on certain mRNAs in an RNA structure-dependent manner. These discoveries indicate that eIF3 may integrate multiple signals to control the translational output of individual mRNAs, much like the Mediator complex in transcription. The aims in this application build on these groundbreaking results to address fundamental questions of how eIF3 functions. We propose to determine the structural basis for eIF3-mediated control of specialized translation, using cryo-electron microscopy (cryo-EM) and in vitro biochemistry. We will also probe how mRNAs regulated by eIF3 are structured in living cells, and dissect the functional importance of these structures to eIF3 regulation and the translational capacity of these mRNAs. Finally, we will use systems biological approaches to explore how eIF3 contributes to the regulation of translation mediated by N-6- methyladenosine (m6A) modifications in mRNAs. By combining advances in cryo-EM to our expertise in human cell engineering, we are in a unique position to unravel the molecular contributions of eIF3 to translation initiation in humans. Taken together, the three aims of this application build on the fundamental insights into eIF3 structure and function obtained in the prior funding period, and address key mechanisms in translational control that could be widespread in human biology. In the long run, our insights into the molecular mechanisms used in human cells to direct eIF3-mediated activation and repression of specific mRNAs could pave the way for the development of new small-molecule and cell-based therapeutics.

项目摘要蛋白质生物合成直接将细胞中的基因型与表型相结合，并且其调节是细胞生物合成的核心。 physiology.我们对蛋白质合成的分子机制的理解经历了一场革命，在过去的十年里，建立在结构生物学和系统生物学的快速发展之上。但重要翻译中的动态事件问题一直没有得到解答，因为这些事件的短暂性这使得它们很难被隔离。在这个应用中，我们建议破译的分子机制，真核生物翻译起始因子3（eIF 3）调节翻译起始。我们最近发现人类eIF 3在翻译中扮演着多重角色。人类eIF 3通常有助于组装翻译在起始密码子处的前起始复合物，但也直接控制特定mRNA的翻译。我们发现eIF 3可以激活或抑制mRNA的翻译，这取决于eIF 3如何与mRNA结合。 mRNA的5'非翻译区（5' UTR）。此外，我们发现eIF 3含有一个亚基EIF 3D，以RNA结构依赖性方式结合某些mRNA上的5 '-m7 G帽。这些发现表明 eIF 3可以整合多种信号来控制单个mRNA的翻译输出，就像转录中的介体复合物。本申请的目的是建立在这些突破性的结果，以解决基本问题， eIF 3功能。我们建议确定eIF 3介导的控制特化细胞的结构基础。翻译，使用冷冻电子显微镜（cryo-EM）和体外生物化学。我们还将探讨如何由eIF 3调节的mRNA在活细胞中被构建，并剖析了这些mRNA的功能重要性。 eIF 3的调控和这些mRNA的翻译能力。最后，我们将使用系统生物学方法来探索eIF 3如何有助于调节N-6- 甲基腺苷（m6 A）修饰。通过将冷冻EM的进步与我们在人类方面的专业知识相结合，细胞工程，我们处于一个独特的位置，以解开eIF 3的分子贡献的翻译人类的启蒙总的来说，这一应用程序的三个目标建立在以下基本见解的基础上： eIF 3的结构和功能在上一个供资期获得，并解决转化的关键机制，这种控制可能在人类生物学中广泛存在。从长远来看，我们对分子机制的了解在人类细胞中用于指导eIF 3介导的特定mRNA的激活和抑制，用于开发新的小分子和基于细胞的疗法。

项目成果

期刊论文数量（21）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Some reassembly required.

DOI：
10.1111/j.1365-2958.2009.07017.x
发表时间：
2010-02
期刊：
Molecular microbiology
影响因子：
3.6
作者：
Cate JH
通讯作者：
Cate JH

Human-like eukaryotic translation initiation factor 3 from Neurospora crassa.

DOI：
10.1371/journal.pone.0078715
发表时间：
2013
期刊：
PloS one
影响因子：
3.7
作者：
Smith MD;Gu Y;Querol-Audí J;Vogan JM;Nitido A;Cate JH
通讯作者：
Cate JH

eIF3 engages with 3'-UTR termini of highly translated mRNAs in neural progenitor cells.

eIF3 与神经祖细胞中高度翻译的 mRNA 的 3-UTR 末端结合。

DOI：
10.1101/2023.11.11.566681
发表时间：
2023
期刊：
bioRxiv : the preprint server for biology
影响因子：
0
作者：
Mestre-Fos,Santi;Ferguson,Lucas;Trinidad,Marena;Ingolia,NicholasT;Cate,JamieHD
通讯作者：
Cate,JamieHD

Assembly of eIF3 Mediated by Mutually Dependent Subunit Insertion.