The use trypanosomes to uncover the identity of proteins, and their precise temporal order of recruitment, required for basal body duplication

中心粒和基体是以微管为基础的细胞器，它们组装纤毛和鞭毛，在真核生物中都是保守的。它们对于在细胞分裂中组织有丝分裂纺锤体以允许染色体分离也是重要的。纤毛/鞭毛用于运动和感觉功能；例如，在精子、肾脏或视网膜细胞中。此外，一组被称为纤毛病的疾病与基础身体复制缺陷有关，导致纤毛/鞭毛的结构受损。对导致这些缺陷的突变的研究为我们提供了关于基本体复制机制的信息。布鲁氏锥虫是一种单细胞原生动物寄生虫，它使用长鞭毛进行运动和感知，因此需要正确组装这个细胞器才能生存。锥虫是研究基底复制和鞭毛伸长的优秀模型；这些过程被很好地控制，并发生在每个细胞周期中。此外，锥虫也会引起人类和牛的疾病。这个项目的目的是利用锥体来揭示蛋白质的身份及其精确的招募时间顺序，这是发生基底复制所必需的。通过使用高通量的基因标签系统，我将对基础身体蛋白质进行大筛选，以确定哪些蛋白质首先复制。这之后将通过耗尽细胞中的蛋白质来表征蛋白质的功能，并看看它是否对基底复制有任何致命的影响。这项研究的发现也可能成为了解其他组织中基底复制过程的相关信息。

Centrioles and basal bodies are microtubule-based organelles that assemble cilia and flagella and are conserved throughout eukaryotes. They are also important for organising mitotic spindles in dividing cells to allow chromosome segregation. Cilia/flagella are used for motility and sensory functions; for example in sperm, kidney or retinal cells. Furthermore, a group of diseases, called ciliopathies, has been linked to defects basal body duplication leading to impaired construction of cilia/flagella. Research on mutations which lead to these defects has provided us with information regarding the mechanism of basal body duplication.Trypanosoma brucei is a single-celled protozoan parasite which uses its long flagellum for motility and sensing, therefore requiring the correct assembly of this organelle to survive. Trypanosomes are excellent models to study basal body duplication and flagellum elongation; these processes are very well controlled and occur in every cell cycle. Moreover, trypanosomes also cause disease in humans and cattle.The aim of this project is to use trypanosomes to uncover the identity of proteins, and their precise temporal order of recruitment, required for basal body duplication to occur. By using a high-throughput gene tagging system, I will carry out a large screen on basal body proteins to identify which proteins duplicate first. This will be followed by characterisation of protein function by depleting the protein from cells and see if it has any lethal effects on basal body duplication. Findings from this study may also become relevant for understanding the process of basal body duplication in other organisms.Priority AreaWUB, ENWW