Dissection of RNA Storage Granules Essential to Plasmodium Transmission

描述(申请人提供)：我的职业目标是成立一个独立的研究小组，通过研究疟疾寄生虫如何准备媒介/宿主传播来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有关疟疾寄生虫的重要问题。获得K22奖将极大地帮助完成建立一个研究小组所需的重要任务，并将提供必要的资金和时间，以积累必要的基础数据和出版物，使其具有竞争力，获得R01研究支持。疟疾寄生虫每年造成3亿至5亿疟疾感染和近100万人死亡。寄生虫的感染需要在蚊子媒介和脊椎动物宿主之间传播，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。最近的工作表明，寄生虫在这两个传播点都使用特定mRNAs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主要集中在通过形成从蚊子传给小鼠的寄生虫子孢子形式的RNA存储颗粒来沉默和保护mRNAs的机制。在这些研究的基础上，这项研究计划将检验特定假设的各个方面，即在媒介/宿主转变处适当形成RNA存储颗粒对疟疾寄生虫的传染性至关重要。为了实现这一点，我建议调查：(1) 通过使用反向遗传学和生化方法剖析Puf2蛋白和IP/MS、RNAseq或候选方法确定的其他蛋白质/RNA相互作用，以及(2)通过有和没有结合伙伴的高分辨率结构和生化研究，研究RNA结合蛋白在RNA存储颗粒形成中的作用。通过成功地实现这些目标，无疑将出现进一步的研究问题，这些问题将进一步集中我们对这些相互作用机制的理解。综上所述，这些发现将突出关键的寄生虫特有特征，可用于新的治疗方法或预防措施。。叙述性疟疾是一个主要的全球卫生负担，每年感染3-5亿人，造成80万人死亡。这项研究将研究在寄生虫从蚊子传播到老鼠的过程中，蛋白质/RNA复合体的形成对寄生虫的传染性至关重要。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干扰它。

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.