Characterizing the Plasmodium falciparum Subpellicular Network

描述恶性疟原虫表膜下网络的特征

• 批准号: 10733433
• 负责人: Ana Karla Cepeda Diaz
• 金额: $ 2.65万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-30 至 2024-05-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10733433
• 关键词: 3-Dimensional; Antimalarials; Architecture; Biochemical; Biogenesis; Biology; Blood; Cell Shape; Cell membrane; Cells; Cessation of life; Complex; Culicidae; Cytoskeletal Proteins; Cytoskeleton; Daughter; Defect; Development; Disease; Drug Targeting; Economics; Electron Microscopy; Erythrocytes; Eukaryota; Family; Filament; Flow Cytometry; Fluorescence-Activated Cell Sorting; Human; Immunofluorescence Immunologic; Individual; Intermediate Filaments; Invaded; Knock-out; Life; Malaria; Mammalian Cell; Maps; Mechanical Stress; Medical; Membrane; Microscopy; Monitor; Morphology; Motor; Names; Organelles; Parasites; Pathway interactions; Pattern; Plasmids; Plasmodium; Plasmodium falciparum; Platinum; Play; Process; Proline; Protein Family; Proteins; Protozoa; Reproduction; Research; Role; Stains; Structure; System; Testing; Toxoplasma gondii; Transmission Electron Microscopy; Valine; asexual; cell motility; daughter cell; drug development; electron tomography; insight; knock-down; live cell microscopy; network architecture; protein function; reconstruction; rhoptry; scaffold; ultra high resolution

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.

摘要 疟原虫的细胞骨架。对人类和人类的复制、运动性和传染性都是必不可少的 蚊子的生活阶段。恶性疟原虫是人类最严重的疟疾的病原体，它利用一种 细胞骨架蛋白家族被称为肺泡蛋白，以满足其多样化的需求。这些中间细丝状 蛋白质不存在于蜂窝状王国之外，这使它们成为有吸引力的药物靶点。肺泡蛋白已经 阶段特定的表达模式，并形成一个复杂的晶格，将寄生虫包裹在膜的正下方， 肺泡菌的另一个特征是由寄生虫的质膜和内膜复合体组成。 在研究肺泡蛋白在伯氏疟原虫蚊虫发育阶段中的作用方面取得了显著进展。 然而，个体肺泡蛋白在疟原虫无性生殖阶段的功能作用仍未被探索。我们有 最近证明，肺泡素PfIMC1g对于恶性疟原虫的无性复制是必不可少的，但它仍然存在。 不清楚这种蛋白质在疟原虫细胞骨架的更大范围内起什么作用。vbl.使用 与超分辨率和白金复制品相结合的可诱导击倒(IKD)和击倒(IKO)系统 在电子显微镜下，我将表征肺泡素PfIMC1g(PF3D7_0525800)在恶性疟原虫子代中的作用 细胞分裂和红细胞入侵。这将阐明其在寄生虫细胞形态和 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的图谱 存在于寄生虫的无性阶段，它们之间的相互作用，并测试它们对 通过传统的KO和IKO方法形成子代细胞。这项研究将增加我们的 对恶性疟原虫SPN的这些构件所起作用和决定作用的理解 它们是如何结合在一起，使其在分割过程中形成、功能和重塑的。加在一起，这些 对中间丝样细胞骨架蛋白及其组织原理的洞察将给我们带来一步 用新的抗疟疾药物更接近于靶向这些蛋白质。