Functional characterization of striated fiber assemblins in malaria parasites

疟疾寄生虫中横纹纤维组装体的功能特征

• 批准号: 10675782
• 负责人: JEFFREY D DVORIN
• 金额: $ 26.55万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-21 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675782
• 关键词: 5 year old; Antimalarials; Apical; Binding Proteins; Biochemical; Biological; Biological Assay; Blood; Cell Nucleus; Cell division; Cell membrane; Cells; Cellular biology; Centrosome; Cessation of life; Child; Clinical; Co-Immunoprecipitations; Complement; Complex; Critical Pathways; Culicidae; Cytokinesis; Cytoplasm; Cytoskeleton; Data; Daughter; Defect; Development; Dimerization; Ensure; Enterobacteria phage P1 Cre recombinase; Erythrocytes; Family; Fiber; Filament; Flagella; Foundations; Future; Generations; Genetic; Genome; Goals; Homologous Gene; Human; Immunofluorescence Immunologic; Individual; Infection; Insecticides; Knock-out; Knowledge; Label; Life Cycle Stages; Location; Malaria; Membrane; Methods; Microscopy; Microtubule-Organizing Center; Microtubules; Molecular; Morbidity - disease rate; Morphology; Nuclear; Organelles; Orthologous Gene; Parasite resistance; Parasites; Pathway interactions; Plasmodium; Plasmodium falciparum; Process; Protein Family; Proteins; Publishing; Reporting; Resistance; Role; Series; Signs and Symptoms; Sirolimus; Structure; Techniques; Testing; Toxoplasma gondii; asexual; combat; daughter cell; genome-wide; improved; kinetosome; knock-down; live cell microscopy; mortality; novel; nuclear division; protein protein interaction

PROJECT SUMMARY Malaria is an important cause of illness and death worldwide, with most of these deaths resulting from Plasmodium falciparum infection. The signs and symptoms of human malaria results from the asexual replication of parasites in human red blood cells. During this clinically important blood stage, P. falciparum parasites divide by schizogony – a process wherein components for several daughter cells are produced within a common cytoplasm and then segmentation, a synchronized cytokinesis, produces individual invasive daughters. The generation of the invasive daughter parasites, known as merozoites, occurs with high fidelity, ensuring that each daughter has a single nucleus and the required organelles. The molecular mechanism underlying this high fidelity of nuclear and organellar partitioning is incompletely understood in Plasmodium. Studies in the related Apicomplexan parasite Toxoplasma gondii identified a family of proteins, referred to as striated fiber assemblins (SFAs), that are critical for parasite cell division. The SFAs are hypothesized to form a connection between the centrosome of dividing nuclei and the newly forming apical ends of the two daughter parasites during T. gondii endodyogeny. The P. falciparum orthologs of the SFAs, PfSFA1 and PfSFA2, have been reported to be dispensable in a genome-wide transposon screen. In contrast to this data, we demonstrate that these two proteins are essential for asexual replication in P. falciparum. Based on our preliminary data, we hypothesize that PfSFA1 and PfSFA2 localize to a filament-like structure during parasite segmentation. In the current proposal, we will test this hypothesize in a series of microscopy assays. Furthermore, we will also evaluate the function of PfSFA1 and PfSFA2 by characterizing parasite arrest and morphologic defects following inducible knockout. To gain a more complete understanding of the molecular functions of PfSFA1 and PfSFA2, we will determine the direct protein interactions by co- immunoprecipitation and the indirect or transient interactions by proximity labeling. By determining a robust protein interaction network for the Plasmodium SFAs, we will establish the foundation to understand their molecular function. We hypothesize that the SFAs proteins are critical for organization of segmentation during the asexual replication of P. falciparum. Together, the proposed studies will interrogate the function of these two essential proteins and directly identify their role during P. falciparum segmentation.

项目摘要 疟疾是全球疾病和死亡的重要原因，其中大多数死亡是由 恶性疟原虫感染。人类疟疾的体征和症状来自无性 人类红细胞中寄生虫的复制。在这个临床上重要的血液阶段，恶性疟原虫 寄生虫通过精神分裂划分 - 产生几个子细胞组件的过程 在常见的细胞质中，然后分割，一种同步的细胞因子，会产生个体 侵入性女儿。侵入性的女儿寄生虫的产生，称为merozoites，发生在 高保真，确保每个女儿都有一个核和所需的细胞器。分子 核和有机分区的高保真度的基础机制尚不完全理解 疟原虫。 在相关的Apicomplexan寄生虫弓形虫弓形虫的研究中确定了一种蛋白质，称为 作为条纹纤维组件（SFA），对寄生虫细胞分裂至关重要。 SFA被认为 在分裂核的中心体与两者的新形成的顶端之间形成联系 gondii内传染性的女儿寄生虫。 SFA，PFSFA1和 据报道，PFSFA2在全基因组的转座子筛选中可处置。与此相反 数据，我们证明，这两种蛋白质对于恶性疟原虫中的无性复制至关重要。基于 在我们的初步数据上，我们假设pfsfa1和pfsfa2局部在类似细丝的结构中， 寄生虫细分。在当前的建议中，我们将在一系列显微镜中测试该假设 测定。此外，我们还将通过表征寄生虫来评估PFSFA1和PFSFA2的功能 诱导敲除之后的逮捕和形态缺陷。为了更完整地了解 PFSFA1和PFSFA2的分子功能，我们将确定共同的直接蛋白质相互作用 免疫沉淀和通过接近标记的间接或瞬态相互作用。通过确定坚固的 疟原虫SFA的蛋白质相互作用网络，我们将建立基础以了解其 分子功能。我们假设SFAS蛋白对于分割的组织至关重要 在恶性疟原虫的无性复制期间。拟议的研究一起审问功能 在这两种基本蛋白质中，并直接识别其在恶性疟原虫分割过程中的作用。