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

中心粒和基小体是基于微管的细胞器，聚集纤毛和鞭毛，并在整个真核生物中保守。它们对于在分裂细胞中组织有丝分裂纺锤体以使染色体分离也很重要。纤毛/鞭毛用于运动和感觉功能；比如精子、肾脏或视网膜细胞。此外，一组疾病，称为纤毛病，已被链接到缺陷的基础体复制导致受损的纤毛/鞭毛的建设。对导致这些缺陷的突变的研究为我们提供了有关基础体复制机制的信息。布鲁氏锥虫是一种单细胞原生动物寄生虫，它利用长长的鞭毛进行运动和感知，因此需要正确组装这种细胞器才能生存。锥虫是研究基体复制和鞭毛伸长的良好模型；这些过程受到很好的控制，发生在每个细胞周期中。此外，锥虫也在人类和牛中引起疾病。这个项目的目的是利用锥虫来揭示蛋白质的身份，以及它们精确的时间顺序，这是发生基础体复制所必需的。通过使用高通量基因标记系统，我将对基础蛋白进行大屏幕筛选，以确定哪些蛋白质首先复制。随后将通过从细胞中消耗蛋白质来表征蛋白质功能，并观察它是否对基础体复制有任何致命影响。这项研究的发现也可能与理解其他生物的基底体复制过程有关。优先区域：awub， ENWW

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