REPEAT-CONTAINING RNA BINDING PROTEINS OF TRYPANOSOMES

锥虫的含有重复序列的 RNA 结合蛋白

• 批准号: 10113510
• 负责人: Inna Afasizheva
• 金额: $ 54.66万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-05 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10113510
• 关键词: 5' Untranslated Regions; Affinity; African; Animals; Antigenic Variation; Architecture; Area; Binding; Binding Sites; Biogenesis; Biology; Biotinylation; Chemicals; Complex; Consensus Sequence; Conserved Sequence; Cryoelectron Microscopy; Custom; Diphosphates; Disease; Economic Burden; Elements; Ensure; Enzymes; Event; Exposure to; Family; Funding; Gene Expression; Genetic; Genetic Materials; Genetic Transcription; Genetic Translation; Goals; Guide RNA; Health Hazards; Helix-Turn-Helix Motifs; Human; Hydrolysis; In Vitro; Individual; Knowledge; Laboratories; Mass Spectrum Analysis; Mastigophora; Mediating; Medical; Messenger RNA; Mitochondria; Mitochondrial RNA; Modality; Modeling; Modification; Molecular; Monitor; Mutate; Mutation; Nuclear; Organelles; Organism; Outcome; Parasites; Parasitic infection; Plants; Poly(A)-Binding Proteins; Polynucleotide Adenylyltransferase; Positioning Attribute; Post-Transcriptional Regulation; Pre-mRNA Polyadenylation Factor; Process; Property; Protein Biosynthesis; Protein Engineering; Proteins; Public Health; RNA; RNA Binding; RNA Degradation; RNA Sequences; RNA-Binding Proteins; Reaction; Reader; Reading; Resolution; Resources; Ribosomes; Role; Science; Site; Specificity; Structure; Substrate Specificity; Tail; Testing; Therapeutic; Transcript; Transcription Initiation; Translational Activation; Translations; Trypanosoma; Trypanosoma brucei brucei; Trypanosomiasis; Vaccines; Validation; X-Ray Crystallography; base; crosslink; endonuclease; gene discovery; health economics; high reward; high risk; in vivo; innovation; insertion/deletion mutation; knock-down; mRNA Precursor; mRNA Stability; mRNA Surveillance; mRNA Transcript Degradation; marginalized population; mitochondrial messenger RNA; new therapeutic target; novel; pathogen; polypeptide; prevent; programs; promoter; reconstitution; recruit; ribosome profiling; sensor; vector control

ABSTRACT Flagellated kinetoplastid protozoans inflict public health hazards and economic burden predominately on marginalized populations in the developing world. The agents of African human and animal trypanosomiasis, Trypanosoma brucei sp., cause some of the most-studied, but least manageable and curable parasitic infections. Failing vector control, lack of vaccines and toxic therapeutics place the onus on identification, molecular understanding and validation of new drug targets. From the fundamental science perspective, these early branching Excavata represent valuable models of mitochondrial RNA biology, antigenic variation, host-pathogen interaction, post-transcriptional regulation, and other processes. Major findings in the previous funding period position a family of RNA binding pentatricopeptide repeat (PPR, 35 amino acids) proteins as the central conduit controlling mitochondrial gene expression. The unique ability of helix-turn-helix repeats to assemble into superhelical arrays recognizing extended and compositionally diverse RNA sequences enables the essential roles of more than 40 PPRs. Often including trypanosome-specific domains, these RNA “sequence readers” commit otherwise non-specific modification enzymes to their substrates, or block RNA degradation and extension. This project will elucidate the mechanisms by which nuclear-encoded PPRs govern mitochondrial mRNA biogenesis and translation via recruiting enzymatic complexes and ribosomes to transcripts at defined processing stages. We establish three priority areas focusing on PPRs that enable mRNA 5′ end modification, 3′ adenylation/uridylation, and translation. Building on our recent discoveries of gene-specific transcription initiation and exonucleolytic pre-mRNA processing, we identified PPR factors acting as “protein cap,” “editing sensor,” poly(A) binding protein, and potential translational activators. Our goal is to dissect the molecular machinery integrating these functions and to decipher principles of RNA recognition by repeat-containing proteins. By elucidating their RNA binding sites, structures, interactions, and outcomes of genetic alteration and protein engineering, this program builds on our momentum in a significant field and expands the knowledge of critical pathogen-specific processes.

抽象的 带鞭毛的动质体原生动物主要造成公共卫生危害和经济负担 发展中国家的边缘化人口。非洲人和动物锥虫病的病原体， 布氏锥虫引起一些研究最多但最难控制和治愈的寄生虫感染。 媒介控制失败、缺乏疫苗和有毒疗法使识别、分子生物学的责任成为可能 新药靶点的理解和验证。从基础科学的角度来看，这些早期 分支 Excavata 代表了线粒体 RNA 生物学、抗原变异、宿主病原体的有价值的模型 相互作用、转录后调控和其他过程。上一资助期间的主要发现 将 RNA 结合五肽重复序列（PPR，35 个氨基酸）蛋白家族定位为中心导管 控制线粒体基因表达。螺旋-转角-螺旋重复序列组装成的独特能力 超螺旋阵列可识别扩展且成分多样的 RNA 序列，从而实现必要的 超过 40 个 PPR 的角色。这些 RNA“序列阅读器”通常包含锥虫特异性结构域 将其他非特异性修饰酶施加到其底物上，或阻止 RNA 降解和 扩大。该项目将阐明核编码的 PPR 控制线粒体的机制 通过将酶复合物和核糖体募集到指定转录物中进行 mRNA 生物发生和翻译 处理阶段。我们建立了三个重点领域，重点关注能够实现 mRNA 5' 末端修饰的 PPR， 3' 腺苷酸化/尿苷酸化和翻译。基于我们最近发现的基因特异性转录 启动和核酸外切前 mRNA 加工，我们确定了 PPR 因子充当“蛋白帽”、“编辑 传感器，“多聚（A）结合蛋白，以及潜在的翻译激活剂。我们的目标是剖析分子 整合这些功能的机器，并通过包含重复序列来破译 RNA 识别原理 蛋白质。通过阐明它们的 RNA 结合位点、结构、相互作用以及遗传改变的结果和 蛋白质工程，该计划建立在我们在一个重要领域的动力基础上，并扩展了我们的知识 关键的病原体特异性过程。