Dissection of RNA Storage Granules Essential to Plasmodium Transmission

疟原虫传播所必需的 RNA 储存颗粒的剖析

• 批准号: 8353932
• 负责人: Scott E Lindner
• 金额: $ 15万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8353932
• 关键词: Address; Affinity Chromatography; Binding; Binding Proteins; Biochemical; Bioinformatics; Blocking Antibodies; C-terminal; Cessation of life; Complex; Crystallization; Culicidae; Cytoplasmic Granules; Data; Dissection; Dominant-Negative Mutation; Epitopes; Escherichia coli; Funding; Genetic; Goals; Immunoprecipitation; Infection; Intervention; Investigation; K22 Award; Ligands; Malaria; Mass Spectrum Analysis; Measures; Medical; Messenger RNA; Methods; Modeling; Mus; Parasites; Parasitology; Plasmodium; Positioning Attribute; Process; Proteins; Proteomics; Publications; RNA; RNA Sequences; RNA-Binding Proteins; RNA-Protein Interaction; Research; Research Proposals; Research Support; Resolution; Reverse Transcriptase Polymerase Chain Reaction; Role; Scientist; Sporozoites; Structure; Testing; Therapeutic Intervention; Time; Training; Transgenic Organisms; Translational Repression; Uncertainty; Vaccines; Work; career; disease transmission; global health; next generation; novel therapeutics; overexpression; positional cloning; protein function; research study; structural biology; three dimensional structure; transcriptomics; transmission process; vector; vector mosquito

DESCRIPTION (provided by applicant): My career goal is to form an independent research group to invest in training the next generation of scientists through addressing how the malaria parasite prepares for vector/host transmission. My training in both structural biology and parasitology has uniquely positioned me to use both approaches to address important questions about the malaria parasite. Receipt of this K22 award will greatly aid in accomplishing the vital tasks required to establish a research group, and would provide the funding and time necessary to amass the foundational data and publications necessary to become competitive for R01 research support. Plasmodium parasites cause 300-500 million malarial infections and nearly a million deaths annually. Infection by the parasite requires the transmission between a mosquito vector and vertebrate host, and vice versa., Targeting these transmission processes has been the focus of many therapeutic interventions, such as through vaccines and antibody blocking strategies. Recent work has demonstrated that the parasite uses translational repression of specific mRNAs during both of these transmission points, and that disrupting this process severely decreases or completely abrogates parasite infectivity. My previous and current studies focus upon the mechanisms of silencing and protecting mRNAs through the formation of RNA storage granules in the sporozoite form of the parasite that are passed from mosquitoes to mice. Building upon these studies, this research proposal will test aspects of the specific hypothesis that proper formation of RNA storage granules at the vector/host transition is crucial for infectivity of the malaria parasite. In order to accomplish this, I propose to investigate: (1) The Role of RNA-Binding Proteins in the Formation of RNA Storage Granules by using reverse genetics and biochemical approaches to dissect the Puf2 protein and other protein/RNA interactions identified by IP/MS, RNAseq or candidate approaches; and (2) The Structure/Function of Storage Granule Components through high-resolution structural and biochemical investigations both with and without their binding partners. By successfully addressing these aims, further research questions will no doubt arise that will further focus our understanding of the mechanisms of these interactions. Taken together, these findings will highlight key parasite-specific features that can be exploited for new therapeutic treatments or preventative measures. . NARRATIVE Malaria is a major global health burden, infecting 300-500 million people and causing 800,000 deaths each year. This research will investigate the formation of protein/RNA complexes that are critical for the infectivity of the parasite as it is passed from a mosquito to a mouse. Understanding these processes will help us to understand disease transmission, and may provide new means to disrupt it.

描述（由申请人提供）：我的职业目标是成立一个独立的研究小组，通过解决疟疾寄生虫如何为媒介/宿主传播做准备，投资于培训下一代科学家。我在结构生物学和寄生虫学方面的训练使我能够使用这两种方法来解决有关疟疾寄生虫的重要问题。获得K22奖将大大有助于完成建立研究小组所需的重要任务，并将提供必要的资金和时间来积累必要的基础数据和出版物，以获得R 01研究支持的竞争力。疟原虫每年造成3 - 5亿疟疾感染和近100万人死亡。寄生虫的感染需要蚊子媒介和脊椎动物宿主之间的传播，反之亦然。针对这些传播过程一直是许多治疗干预措施的重点，例如通过疫苗和抗体阻断策略。最近的研究表明，寄生虫在这两个传播点使用特定mRNA的翻译抑制，破坏这一过程严重降低或完全消除寄生虫的感染性。我以前和现在的研究集中在沉默和保护mRNAs的机制，通过形成RNA存储颗粒的寄生虫的子孢子形式，从蚊子传递到小鼠。在这些研究的基础上，这项研究计划将测试特定假设的各个方面，即在载体/宿主过渡期正确形成RNA储存颗粒对疟原虫的感染性至关重要。为了实现这一点，我建议调查：（1） RNA结合蛋白在RNA储存颗粒形成中的作用，通过使用反向遗传学和生物化学方法来剖析Puf 2蛋白和通过IP/MS、RNAseq或候选方法鉴定的其他蛋白/RNA相互作用;以及（2）通过高分辨率结构和生物化学研究储存颗粒组分的结构/功能，包括有和没有它们的结合伴侣。通过成功地解决这些目标，进一步的研究问题无疑会出现，这将进一步集中我们对这些相互作用机制的理解。总之，这些发现将突出可用于新的治疗或预防措施的关键寄生虫特异性特征。.疟疾是一个主要的全球健康负担，每年感染3亿至5亿人，造成80万人死亡。这项研究将研究蛋白质/RNA复合物的形成，这些复合物对寄生虫的感染性至关重要，因为它是从蚊子传给小鼠的。了解这些过程将有助于我们了解疾病的传播，并可能提供新的手段来破坏它。