Genome-wide subcellular protein map for the flagellate parasite Trypanosoma brucei.
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.
Acknowledgements: We thank the Wellcome Trust for funding through Investigator Awards (104627/Z/14/Z, K.G.; 217138/Z/19/Z, M.C.) a Biomedical Resource Grant (108445/Z/15/Z, K.G.), a Sir Henry Wellcome and Sir Henry Dale Fellowship (211075/Z/18/Z and 103261/Z/13/Z, R.W.) and a Biomedical Resource Grant supporting VEuPathDB (218288/Z/19/Z, C.H.F.). We also thank the TrypTag scientific advisory group for their support and advice.