Characterizing the Plasmodium falciparum Subpellicular Network

摘要 疟原虫的细胞骨架。对人类和哺乳动物的复制、运动和感染性至关重要， 蚊子的生活阶段恶性疟原虫是最严重的人类疟疾的病原体， 细胞骨架蛋白家族称为肺泡蛋白，以满足其不同的需求。这些中间的类似于 蛋白质在肺泡王国之外不存在，使它们成为有吸引力的药物靶点。肺泡蛋白 阶段特异性表达模式，并形成一个复杂的晶格，将寄生虫包裹在表膜下方， 由寄生物质膜和内膜复合体组成的泡孔动物的另一个标志。 在描述肺泡蛋白在伯氏疟原虫蚊子阶段中的作用方面已经取得了显着进展。 然而，在疟原虫无性阶段的单个肺泡蛋白的功能作用仍然没有得到探索。我们有 最近证明，肺泡蛋白PfIMC 1g是恶性疟原虫无性复制所必需的，但它仍然存在， 尚不清楚这种蛋白质在疟原虫细胞骨架的更大背景下起什么作用。使用 诱导型敲除（iKD）和敲除（iKO）系统结合超分辨率和铂复制品 电镜下，我将表征肺泡蛋白PfIMC 1g（PF3D7_0525800）在恶性疟原虫子体中的作用 细胞分裂和红细胞侵入。这将阐明其在寄生虫细胞形状中的个体作用， 承受机械压力的能力。此外，我将使用扩张显微镜来绘制其他肺泡蛋白 存在于寄生虫的无性阶段，它们彼此之间的相互作用，并测试它们的必要性， 通过常规KO和iKO方法形成子细胞。这项研究将提高我们的 了解恶性疟原虫SPN的这些组成部分的功能作用， 它们如何在分割过程中结合在一起，使其形成，功能和重塑。所有这些 对中间丝样细胞骨架蛋白及其组织原理的深入了解将使我们更进一步 用新的抗疟疾药物来靶向这些蛋白质。

ABSTRACT The cytoskeleton of Plasmodium spp. is essential for replication, motility, and infectivity in both human and mosquito life stages. P. falciparum, the causative agent of the most severe form of human malaria, leverages a family of cytoskeletal proteins known as the alveolins to meet its diverse needs. These intermediate filament-like proteins are absent outside the Alveolate kingdom, making them attractive drug targets. The alveolins have stage-specific expression patterns and form an intricate lattice which envelops the parasite just below the pellicle, another hallmark of Alveolata consisting of the parasite plasma membrane and inner membrane complex. Remarkable progress has been achieved in characterizing the role of alveolins in P. berghei mosquito stages. However, the functional role of individual alveolins in Plasmodium asexual stages remains unexplored. We have recently demonstrated that the alveolin PfIMC1g is essential for P. falciparum asexual replication, but it remains unclear what function this protein serves within the larger context of the Plasmodium cytoskeleton. Using inducible knockdown (iKD) and knockout (iKO) systems in conjunction with super-resolution and platinum replica electron microscopy, I will characterize the role of alveolin PfIMC1g (PF3D7_0525800) in P. falciparum daughter cell segmentation and red blood cell invasion. This will elucidate its individual role in parasite cell shape and ability to endure mechanical stress. In addition, I will use expansion microscopy to map the other alveolins present in the asexual stages of the parasite, their interactions with each other, and test their essentiality for daughter cell formation through conventional KO and iKO approaches. This research will increase our understanding of the functional role that these building blocks of the P. falciparum SPN serve and determine how they come together to enable its formation, function, and remodeling during segmentation. Together, these insights into intermediate-filament-like cytoskeletal proteins and their organizing principles will bring us one step closer to targeting these proteins with new antimalarial drugs.