Shining a light on dense granules- biochemical, genetic and cell biological investigation of an essential but understudied compartment in malarial -

研究问题：拟建项目旨在利用跨学科技术研究人类最致命的疟疾寄生虫——顶复合体疟原虫恶性疟原虫的致密颗粒形成和蛋白靶向。分子生物学方法和成像技术将用于鉴定靶蛋白的密集颗粒的基因序列。一旦这些信号被确定，生物信息学方法将用于预测新的致密颗粒蛋白。这种生物信息学技术的使用符合MRC LID数据分析，信息学和计算方法的关键技能。然后可以使用质谱法确认这些蛋白质的存在，并通过观察使用CRISPR/Cas9基因组编辑技术产生的敲除突变体来确定它们的功能。理由：致密颗粒是疟原虫感染宿主红细胞所必需的。由于疟疾的衰弱症状是寄生虫入侵并在宿主红细胞内存活的产物，阐明这一过程的机制并有助于理解疟原虫系统生物学将为医学科学提供非常重要的信息。此外，该项目还可以鉴定适合用于抑制剂筛选的蛋白质，以期发现能够抑制寄生虫生长的化合物。背景：疟疾是由疟原虫属顶复体寄生虫引起的一种重要传染病，2016年全球有2.16亿病例，估计有44.5万例死亡，其中65%的病例发生在15岁以下的人群中。然而，防治疟疾的进展正在下降，因此开发新的干预措施至关重要。疟疾的症状是当寄生虫疟原虫（zoite）的感染生命阶段侵入宿主红细胞并形成寄生液泡（PV）时引起的，寄生虫可以在其中繁殖。和其他顶复合体寄生物一样，疟原虫通过zoite侵入宿主细胞需要三个专门的分泌细胞器的功能；微素、红体和致密颗粒（DG）。这三种细胞器已被证明对宿主细胞入侵和PV形成至关重要，而在疟原虫的宿主细胞入侵中，红素和微素起着至关重要的作用。然而，尽管它们在寄生虫生存中起着至关重要的作用，但DG在疟原虫宿主细胞感染中的作用机制仍不清楚。

- parasitesResearch Questions: The proposed project aims to use interdisciplinary techniques to investigate dense granule formation and protein targeting in the apicomplexan parasite Plasmodium falciparum, the most lethal of the human malaria parasites. Molecular biological approaches and imaging techniques will be used to identify genetic sequences which target proteins to the dense granules. Once these signals have been identified, bioinformatics approaches will be used to predict novel dense granule proteins. The use of such bioinformatics techniques meets the MRC LID key skills of data analytics, informatics and computational approaches. The presence of these proteins can then be confirmed using mass spectrometry, and their function determined by observing knock-out mutants generated using CRISPR/Cas9 genome editing technology. Rationale: Dense granules are essential for Plasmodium infection of the host erythrocyte. Since the debilitating symptoms of malaria are a product of parasite invasion and survival within the host erythrocytes, elucidating the mechanisms which underly this process and contributing to the understanding of Plasmodium systems biology will provide information of great importance to medical science. Additionally, this project could identify proteins suitable for inhibitor screening with a view to discovering compounds capable of inhibiting parasite growth.Background: Malaria is a significant infectious disease caused by apicomplexan parasites of the genus Plasmodium with 216 million cases and an estimated 445,000 mortalities worldwide in 2016 , and 65% of cases occurring in those under the age of 15. However, advances against malaria are in decline, therefore the development of novel interventions is vital.The symptoms of malaria are caused when the infective life stage of the parasite Plasmodium (the zoite), invades the host erythrocyte and forms a parasitophorous vacuole (PV) in which the parasite can reproduce. As in other apicomplexan parasites, Plasmodium spp. invasion of the host cell by the zoite requires the function of three specialised secretory organelles; micronemes, rhoptries, and dense granules (DG). These three organelles have been demonstrated to be essential to host cell invasion and PV formation, with rhoptries and micronemes holding essential roles in host cell invasion in Plasmodium. However, despite their vital role in parasite survival, the mechanisms of DG function in Plasmodium host cell infection remain largely unknown.