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RAPs-mediated post-transcriptional control in Apicomplexan parasites

RAP 介导的顶复门寄生虫转录后控制

基本信息

批准号：
10466864
负责人：
Karine Gaelle Le Roch
金额：
$ 56.06万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-19 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10466864
关键词：
Address Antimalarials Antiparasitic Agents Aptamer Technology Binding Proteins Biochemical Biogenesis Bioinformatics Biology CRISPR/Cas technology Cell physiology Cessation of life Characteristics Code Communicable Diseases Computer Analysis Data Development Drug Targeting Engineering Epitopes Erythrocytes Eukaryota Family Future Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Transcription Genomics Goals Health High-Throughput Nucleotide Sequencing Human In Situ Knowledge Lead Life Cycle Stages Maintenance Malaria Mass Spectrum Analysis Mediating Messenger RNA Molecular Morbidity - disease rate Nuclear Nuclear Export Organelles Organism Parasites Pathway interactions Peptide Elongation Factor 2 Phase Phenotype Plasmodium Plasmodium falciparum Play Post-Transcriptional Regulation Protein Binding Domain Protein Biosynthesis Protein Family Proteins Proteomics Publishing RNA RNA Recognition Motif RNA Splicing RNA-Binding Proteins Research Resistance Ribonucleoproteins Role Sex Differentiation Technology Therapeutic Therapeutic Intervention Time Transcript Translations Vaccines Validation Virulence base comparative crosslinking and immunoprecipitation sequencing design drug discovery experimental study genome-wide human pathogen improved insight mRNA Stability mortality multidisciplinary new therapeutic target novel novel therapeutic intervention novel therapeutics pre-clinical programs protein expression small molecule inhibitor transcriptome transcriptome sequencing zygote

项目摘要

Project Summary/Abstract In the malaria parasite, gene regulation is heavily dependent on mechanisms acting at the post- transcriptional and translational levels. More importantly, Plasmodium possesses many atypical characteristics in both pathways when compared to its human host. These unusual features hence extend our therapeutic window against this deadly parasite, yet these characteristics have been poorly explored at the molecular level. The main goal of this project is to characterize the role of newly identified apicomplexan-specific RNA- binding proteins {RAPs) in parasite development and virulence, and to validate their potential as novel drug targets. The fact that Plasmodium parasites modulate part of his gene expression at the post-transcriptional level was inspired by many early studies that demonstrated that the global proteomic profiles often lagged behind the transcriptional profiles. Our newly published large-scale bioinformatics reinforce this conjecture as we determined that at least 18% of all coding-gene are predicted to be potential RNA binding domain-containing protein (RBP). Furthermore, many of these parasite RBPs were experimentally shown to interact in situ with mRNAs. In all eukaryotic organisms, RBPs are essential to regulate mRNA processing at multiple levels including splicing, transport, mRNA stability and turnover, as well as mRNA localization and translational efficiency. In the human malaria parasite, very few RBPs have been characterized. We propose to address this knowledge gap by examining the function of the parasite specific RNA-binding domain proteins abundant in Apicomplexans, or RAPs. More specifically, we will use state-of-the-art genomics, molecular, and cellular approaches to determine the role of the 21 identified RAPs in parasite development and survival. In Aim 1, we will characterize the essentiality, subcellular localization and phenotypes of RAPs in P. falciparum across different developmental stage using novel CRISPR/Cas9 approaches. In Aim 2, we will define the global RAP- related ribonucleoprotein interaction network across various parasite developmental stages by developing high-throughput sequencing technologies and pull-down strategies followed by mass spectrometry analysis. Our complementary approaches will not only identify key specific RBPs contributing to parasite development, but also uncover unique post-transcriptional networks in a eukaryotic human pathogen. By providing fundamental insights into mechanisms regulating translation in Plasmodium, this project will improve our ability to design new drugs and novel lines of defense against malaria and many other infectious disease agents.

项目概要/摘要在疟原虫中，基因调控很大程度上依赖于作用于转录后和翻译水平。更重要的是，Plasmodium 拥有许多与人类宿主相比，这两种途径均具有非典型特征。这些不寻常的因此，这些功能扩展了我们针对这种致命寄生虫的治疗窗口，但这些在分子水平上对这些特征的探索很少。该项目的主要目标是表征新鉴定的 apicomplexan 特异性 RNA 结合蛋白 (RAP) 在寄生虫的发育和毒力，并验证其作为新药物靶点的潜力。事实上，疟原虫寄生虫在转录后调节其部分基因表达水平受到许多早期研究的启发，这些研究表明全球蛋白质组谱通常落后于转录谱。我们新发表的大规模生物信息学强化了这个猜想，因为我们确定至少 18% 的编码基因是被预测的是潜在的含有RNA结合域的蛋白质（RBP）。此外，许多这些寄生虫实验表明 RBP 可以与 mRNA 原位相互作用。在所有真核生物中， RBP 对于调节 mRNA 的多个水平加工至关重要，包括剪接、运输、mRNA 稳定性和周转以及 mRNA 定位和翻译效率。在人类疟原虫中，很少有 RBP 被表征。我们建议通过检查寄生虫特异性 RNA 结合的功能来弥补这一知识差距顶复门 (Apicomplexans) 或 RAP 中丰富的结构域蛋白。更具体地说，我们将使用最先进的基因组学、分子和细胞方法来确定 21 确定了寄生虫发育和生存中的 RAP。在目标 1 中，我们将描述恶性疟原虫中 RAP 的重要性、亚细胞定位和表型使用新型 CRISPR/Cas9 方法的不同发育阶段。在目标 2 中，我们将定义跨各种寄生虫的全局 RAP 相关核糖核蛋白相互作用网络通过开发高通量测序技术和 Pull-down 来应对不同的发展阶段策略随后进行质谱分析。我们的互补方法不仅将确定有助于寄生虫发育的关键特定 RBP，同时也发现独特的 RBP 真核人类病原体的转录后网络。通过提供对疟原虫翻译调节机制的基本见解，该项目将提高我们设计新药和针对疟疾和许多其他疾病的新防线的能力传染病媒介。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Functional genomics of RAP proteins and their role in mitoribosome regulation in Plasmodium falciparum.

DOI：
10.1038/s41467-022-28981-7
发表时间：
2022-03-11
期刊：
Nature communications
影响因子：
16.6
作者：
Hollin T;Abel S;Falla A;Pasaje CFA;Bhatia A;Hur M;Kirkwood JS;Saraf A;Prudhomme J;De Souza A;Florens L;Niles JC;Le Roch KG
通讯作者：
Le Roch KG

Genome-Wide Analysis of RNA-Protein Interactions in Plasmodium falciparum Using eCLIP-Seq.