Biochemical and Functional Analysis of Trypanin

描述(申请人提供)：非洲锥虫(例如布鲁氏锥虫)是引起非洲昏睡病的原生动物寄生虫，这是一种致命的疾病，具有毁灭性的健康和经济影响。这些寄生虫是双基因生物，它们的部分生命周期在哺乳动物宿主中度过，部分在昆虫媒介(采采蝇)中度过。锥虫在生命周期的两个阶段都有很高的运动性，运动性是寄生虫发展和疾病发病的核心。单个鞭毛是一种重要的多功能细胞器，在细胞运动、寄主-寄生虫相互作用、细胞形态发生和细胞分裂中起着关键作用。令人惊讶的是，我们对分子水平上的鞭毛器知之甚少。特别是，我们缺乏对鞭毛蛋白如何在轴丝内组装成超分子结构，以及它们如何单独和共同作用来驱动细胞运动和其他鞭毛功能的了解。这项研究的长期目标是促进我们对锥虫鞭毛的了解，并利用锥虫作为研究真核纤毛的模型。这将使用功能和结构方法相结合的方法来完成。将使用RNAi、定点突变和超微结构分析来研究胰酶和动力蛋白调节复合体(DRC)的其他成分的功能。DRC是调节鞭毛运动的信号转导途径的一部分，在血流形式的锥虫中是必不可少的，使其成为候选的药物靶点。锥虫的鞭毛类似于包括人类在内的其他真核生物中的纤毛和鞭毛。鞭毛是几种人类病原体运动所必需的，存在于人体的大多数组织中。它们执行运动、运输和感觉功能。需要纤毛的病原体引起的传染病包括非洲昏睡病和疟疾。这些疾病加在一起造成了全世界约4亿人的死亡和发病。由纤毛缺陷引起的人类遗传性疾病包括：脑积水、不孕症、癫痫、左右轴缺陷、眼睛疾病、多囊肾病和肥胖。因此，除了解决细胞生物学中的基本问题外，这项研究还直接影响到理解和治疗人类传染病和遗传病的努力。与公共卫生相关：非洲锥虫是导致非洲昏睡病的原生动物寄生虫，这是一种致命的疾病，具有毁灭性的健康和经济影响。锥虫鞭毛是生存所必需的，也是寄生虫发育和疾病发病的中心。因此，这项研究将直接影响人们对传染性人类疾病的认识和治疗。

DESCRIPTION (provided by applicant): African trypanosomes (e.g. Trypanosoma brucei) are protozoan parasites that cause African sleeping sickness, a fatal disease with devastating health and economic impact. These parasites are digenetic organisms, spending part of their life cycle in a mammalian host and part in an insect vector (the tsetse fly). Trypanosomes are highly motile in both life cycle stages and motility is central to parasite development and disease pathogenesis. Motility is mediated by a single flagellum that is an essential and multifunctional organelle with critical roles in cell motility, host-parasite interaction, cell morphogenesis and cell division. Surprisingly, we know very little about the flagellar apparatus at the molecular level. In particular, we lack an understanding of how flagellar proteins are assembled into supramolecular structures within the axoneme and how they function individually and collectively to drive cell motility and other flagellum functions. The long-term goal of the proposed research is to advance our understanding of the trypanosome flagellum and to exploit trypanosomes as a model to investigate the eukaryotic cilium. This will be done using a combination of functional and structural approaches. RNAi, site-directed mutagenesis and ultrastructural analyses will be used to investigate the function of trypanin and other components of the dynein regulatory complex (DRC). The DRC is part of a signal transduction pathway that regulates flagellar motility and is essential in bloodstream-form trypanosomes, making it a candidate drug target. The trypanosome flagellum is analogous to cilia and flagella in other eukaryotes, including humans. Flagella are required for motility of several human pathogens and are present on most tissues of the human body. They perform motility, transport and sensory functions. Infectious diseases caused by pathogens that require cilia include African sleeping sickness and Malaria. Together, these diseases are responsible for mortality and morbidity in approximately 0.4 billion people world-wide. Heritable human diseases caused by cilia defects include: hydrocephalus, infertility, epilepsy, left-right axis defects, eye disorders, polycystic kidney disease and obesity. Therefore, in addition to addressing fundamental questions in cell biology, this research directly impacts efforts to understand and treat infectious diseases and genetic diseases in humans. PUBLIC HEALTH RELEVANCE: African trypanosomes are protozoan parasites that cause African sleeping sickness, a fatal disease with devastating health and economic impact. The trypanosome flagellum is essential for viability and is central to parasite development and disease pathogenesis. Therefore, this research will directly impact efforts to understand and treat infectious human disease.