Biochemical and Functional Analysis of Trypanin

描述（由申请人提供）：非洲锥虫（如布鲁氏锥虫）是引起非洲昏睡病的原生动物寄生虫，这是一种具有破坏性健康和经济影响的致命疾病。这些寄生虫是遗传生物，其生命周期的一部分在哺乳动物宿主中度过，一部分在昆虫载体（采采蝇）中度过。锥虫在生命周期的两个阶段都具有高度的运动性，运动性是寄生虫发育和疾病发病的核心。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，我们在分子水平上对鞭毛器官所知甚少。特别是，我们缺乏对鞭毛蛋白如何在轴突内组装成超分子结构以及它们如何单独和集体地驱动细胞运动和其他鞭毛功能的理解。本研究的长期目标是提高我们对锥虫鞭毛的认识，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。RNAi、位点定向诱变和超微结构分析将用于研究锥虫蛋白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，在血流型锥虫中是必不可少的，使其成为候选药物靶点。锥虫鞭毛类似于其他真核生物（包括人类）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。它们具有运动、运输和感觉功能。由需要纤毛的病原体引起的传染病包括非洲昏睡病和疟疾。这些疾病加起来造成全世界约4亿人死亡和发病。由纤毛缺陷引起的遗传性人类疾病包括：脑积水、不孕症、癫痫、左右轴缺陷、眼疾、多囊肾病和肥胖。因此，除了解决细胞生物学的基本问题外，这项研究还直接影响了理解和治疗人类传染病和遗传疾病的努力。公共卫生相关性：非洲锥虫是引起非洲昏睡病的原生动物寄生虫，这是一种对健康和经济造成毁灭性影响的致命疾病。锥虫鞭毛对生存能力至关重要，是寄生虫发育和疾病发病的核心。因此，这项研究将直接影响了解和治疗人类传染性疾病的努力。

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.