Guide RNA Binding Complex

引导RNA结合复合物

基本信息

批准号：
10183140
负责人：
Ruslan Afasizhev
金额：
$ 49.92万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-05-10 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10183140
关键词：
Antigenic Variation Antisense RNA Architecture Area Binding Binding Sites Biology Characteristics Chemicals Complex Cryoelectron Microscopy Crystallization DNA DNA-Directed RNA Polymerase Data Development Diphosphates Ensure Event Excision Exonuclease Funding Gap Junctions Genes Genetic Genetic Transcription Genome Goals Guide RNA Health Hazards Holoenzymes Hybrids Hydrolysis Individual Investigation Kinetoplast DNA Knowledge Maps Mass Spectrum Analysis Mediator of activation protein Medical Messenger RNA Methods Mitochondria Mitochondrial RNA Modality Modeling Modification Molecular Parasites Pathway interactions Pattern Polyadenylation Polynucleotide Adenylyltransferase Positioning Attribute Pre-mRNA Polyadenylation Factor Process Property Proteins RNA RNA Binding RNA Editing RNA Processing Reaction Research Resolution Ribosomes Role Specificity Structure System Tail Testing Transcript Transcription Initiation Site Translational Activation Translations Tropical Disease Trypanosoma Trypanosoma brucei brucei base crosslink endonuclease experimental study genome-wide analysis health economics in vivo insertion/deletion mutation mRNA Precursor mRNA Stability marginalized population multimodality new therapeutic target novel pathogen polypeptide programs promoter reconstitution recruit sensor transcriptome tripolyphosphate

项目摘要

ABSTRACT Trypanosoma brucei species inflict health hazards and economic hardship on arguably the most marginalized populations in the world. Some of the best-studied Excavata, trypanosomes also represent important models in many areas of research, including antigenic variation, host-pathogen interaction, developmental reprogramming and mitochondrial biology. This project will elucidate mechanisms by which macromolecular RNA editing substrate binding complex (RESC) stabilizes and delivers mitochondrial pre-mRNAs and guide RNAs into the U-insertion/deletion editing pathway, and coordinates polyadenylation and translation of edited mRNAs. We establish the RESC platform as the RNA binding constituent of the editing holoenzyme and seek to investigate its role in editing reactions, and functions beyond the RNA editing process. To this end, we demonstrate that RESC-associated MERS1 pyrophosphohydrolase and KPAP1 poly(A) polymerase target pre-mRNA 5′ and 3′ ends, respectively. Importantly, both 5′ pyrophosphate removal and 3′ A-tailing appear to be critical for pre-mRNA stabilization prior to editing. Conversely, specific module within RESC is suggested to couple the completion of editing with post-editing 3′ A/U-tailing and mRNA binding to the ribosome. Collectively, the existing evidence positions the ~25 polypeptide RESC complex as the multimodal nexus of mitochondrial RNA processing. Furthermore, initial investigation of RESC-associated MERS1 complex, RNA polymerase (MTRNAP), and the 3′ processome (MPsome) challenges the long-standing model of multicistronic maxicircle transcription and endonucleolytic partitioning of primary transcripts. The proposed experiments will deepen understanding of RNA editing by determining the RESC structure at near-atomic resolution and RNA binding specificities of individual subunits. We will test a broad functional hypothesis that discrete RESC modules coordinate completion of mRNA editing with 3′ modification and translational activation. Finally, we put forward a fundamentally novel concept of monocistronic pre-mRNAs that are transcribed from individual promoters and shaped by 5ʹ modification and antisense RNA-controlled 3′-5′ degradation. By elucidating the RESC structure, RNA binding properties, and higher-order interactions, and evaluating the paradigm-shifting “monocistronic hypothesis,” this program will expand the knowledge of critical parasite-specific processes and may provide new drug targets.

抽象的布氏锥虫对最有争议的最边缘群体造成健康危害和经济困难世界上一些研究最深入的锥虫也代表了重要的模型。许多研究领域，包括抗原变异、宿主-病原体相互作用、发育重编程该项目将阐明大分子 RNA 编辑的机制。底物结合复合物 (RESC) 可稳定线粒体前体 mRNA，并将其引导至线粒体中 U-插入/删除编辑途径，并协调编辑的 mRNA 的聚腺苷酸化和翻译。建立RESC平台作为编辑全酶的RNA结合成分并寻求研究它在编辑反应中的作用以及 RNA 编辑过程之外的功能为此，我们证明了这一点。 RESC 相关的 MERS1 焦磷酸水解酶和 KPAP1 聚 (A) 聚合酶靶向前 mRNA 5' 和 3' 重要的是，5' 焦磷酸去除和 3' A 尾部似乎对前 mRNA 至关重要。离线编辑之前的稳定，建议使用RESC中的特定模块来完成编辑后的 3' A/U 尾部和 mRNA 与核糖体的结合总体上是现有的证据。将 ~25 多肽 RESC 复合体定位为线粒体 RNA 加工的多模式连接。此外，对 RESC 相关 MERS1 复合物、RNA 聚合酶 (MTRNAP) 和 3’ 的初步研究 processome (MPsome) 挑战了长期存在的多顺反子巨环转录模型所提出的实验将加深对 RNA 的理解。通过确定近原子分辨率的RESC结构和个体的RNA结合特异性进行编辑我们将测试一个广泛的功能假设，即离散的 RESC 模块协调 mRNA 的完成。最后，我们提出了一个全新的概念：3′修饰和翻译激活。单顺反子前体 mRNA 由单个启动子转录并通过 5ʹ 修饰形成反义 RNA 控制的 3'-5' 降解通过阐明 RESC 结构、RNA 结合特性和高阶相互作用，并评估范式转换的“单顺反子假说”，该程序将扩大对关键寄生虫特定过程的了解，并可能提供新的药物靶点。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

RNA Editing TUTase 1: structural foundation of substrate recognition, complex interactions and drug targeting.

DOI：
10.1093/nar/gkw917
发表时间：
2016-12-15
期刊：
Nucleic acids research
影响因子：
14.9
作者：
Rajappa-Titu L;Suematsu T;Munoz-Tello P;Long M;Demir Ö;Cheng KJ;Stagno JR;Luecke H;Amaro RE;Aphasizheva I;Aphasizhev R;Thore S
通讯作者：
Thore S

U-insertion/deletion RNA editing multiprotein complexes and mitochondrial ribosomes in Leishmania tarentolae are located in antipodal nodes adjacent to the kinetoplast DNA.

利什曼原虫中的 U 插入/删除 RNA 编辑多蛋白复合物和线粒体核糖体位于与动质体 DNA 相邻的反足节中。

DOI：
10.1016/j.mito.2015.10.006
发表时间：
2015
期刊：
Mitochondrion
影响因子：
4.4
作者：
Wong,RichardG;Kazane,Katelynn;Maslov,DmitriA;Rogers,Kestrel;Aphasizhev,Ruslan;Simpson,Larry
通讯作者：
Simpson,Larry

Mitochondrial RNA editing in trypanosomes: small RNAs in control.