Dissection of RNA Storage Granules Essential to Plasmodium Transmission

描述（由申请人提供）：我的职业目标是组建一个独立的研究小组，通过研究疟疾寄生虫如何为媒介/宿主传播做准备，投资培训下一代科学家。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关于疟疾寄生虫的重要问题。获得这一K22奖将极大地帮助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R01研究支持的竞争力。疟原虫每年导致3 -5亿人感染疟疾，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都使用特定mrna的翻译抑制，破坏这一过程会严重降低或完全消除寄生虫的传染性。我以前和现在的研究重点是通过在蚊子传播给小鼠的寄生虫的孢子体形式中形成RNA储存颗粒来沉默和保护mrna的机制。在这些研究的基础上，本研究计划将测试特定假设的各个方面，即在载体/宿主过渡过程中RNA储存颗粒的适当形成对疟疾寄生虫的传染性至关重要。为了做到这一点，我建议进行以下调查：(1)

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.