Sir William Dunn School of Pathology, University of Oxford, Oxford, UK.
Department of Biological and Medical Sciences, Oxford Brookes University, Oxford, UK.
Nat Microbiol. 2023 Mar;8(3):533-547. doi: 10.1038/s41564-022-01295-6. Epub 2023 Feb 20.
Trypanosoma brucei is a model trypanosomatid, an important group of human, animal and plant unicellular parasites. Understanding their complex cell architecture and life cycle is challenging because, as with most eukaryotic microbes, ~50% of genome-encoded proteins have completely unknown functions. Here, using fluorescence microscopy and cell lines expressing endogenously tagged proteins, we mapped the subcellular localization of 89% of the T. brucei proteome, a resource we call TrypTag. We provide clues to function and define lineage-specific organelle adaptations for parasitism, mapping the ultraconserved cellular architecture of eukaryotes, including the first comprehensive 'cartographic' analysis of the eukaryotic flagellum, which is vital for morphogenesis and pathology. To demonstrate the power of this resource, we identify novel organelle subdomains and changes in molecular composition through the cell cycle. TrypTag is a transformative resource, important for hypothesis generation for both eukaryotic evolutionary molecular cell biology and fundamental parasite cell biology.
布氏锥虫是一种模式原生动物，是一类重要的人类、动物和植物单细胞寄生虫。由于与大多数真核微生物一样，约 50%的基因组编码蛋白的功能完全未知，因此理解其复杂的细胞结构和生命周期具有挑战性。在这里，我们使用荧光显微镜和表达内源性标记蛋白的细胞系，绘制了 89%的 T. brucei 蛋白质组的亚细胞定位图谱，我们称之为 TrypTag。我们提供了功能线索，并定义了寄生虫特有的细胞器适应性，绘制了真核生物的超保守细胞结构图谱，包括对真核鞭毛的首次全面“制图”分析，鞭毛对于形态发生和病理学至关重要。为了展示该资源的强大功能，我们通过细胞周期鉴定了新的细胞器亚区和分子组成的变化。TrypTag 是一个变革性的资源，对于真核进化分子细胞生物学和寄生虫基础细胞生物学的假说生成都非常重要。
