RAPs-mediated post-transcriptional control in Apicomplexan parasites

RAP 介导的顶复门寄生虫转录后控制

• 批准号: 9788270
• 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
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-19 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9788270
• 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 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; 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

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.

项目摘要/摘要 在疟疾寄生虫中，基因调节在很大程度上取决于作用在 转录后和翻译水平。更重要的是，疟原虫拥有许多 与人类宿主相比，这两种途径的非典型特征。这些不寻常 因此，特征将我们的治疗窗口扩展到这个致命的寄生虫，但是这些 特征在分子水平上的探索很差。这个项目的主要目标是 表征新鉴定的Apicomplexan特异性RNA结合蛋白（RAP）的作用 寄生虫发育和毒力，并验证其作为新型药物靶标的潜力。 疟原虫寄生虫在转录后调节其基因表达的一部分 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学特征经常 落后于转录轮廓。我们新发表的大规模生物信息学 加强这种猜想，因为我们确定所有编码基因的至少18％被预测 成为潜在的含RNA结合结构域的蛋白（RBP）。此外，其中许多寄生虫 实验表明RBP与mRNA相互作用。在所有真核生物中， RBP对于在包括拼接的多个级别调节mRNA处理至关重要， 运输，mRNA稳定性和周转率以及mRNA定位和翻译 效率。在人类疟疾的寄生虫中，很少有RBP的特征。我们建议 通过检查寄生虫特异性RNA结合的功能来解决此知识差距 Apicomplexans或Raps中丰富的域蛋白质。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的作用 确定了寄生虫发育和生存中的说唱。在AIM 1中，我们将表征 遍布恶性疟原虫中的大量的大量，亚细胞定位和表型 使用新颖的CRISPR/CAS9方法的不同发育阶段。在AIM 2中，我们将定义 跨各种寄生虫的全球RAP-与核糖核蛋白相互作用网络 通过开发高通量测序技术和下拉的发展阶段 策略遵循质谱分析。我们的补充方法不仅会 确定特定特定的RBP，促进寄生虫开发，但也发现了独特的 真核人类病原体中的转录后网络。通过提供 对调节质量翻译的机制的基本见解，该项目将 提高我们设计新药物和针对疟疾和许多其他许多其他防御措施的能力 传染病药物。