title: Conservation and divergence within the clathrin interactome of Trypanosoma cruzi.
abstract: Trypanosomatids are parasitic protozoa with a significant burden on human health. African and American trypanosomes are causative agents of Nagana and Chagas disease respectively, and speciated about 300 million years ago. These parasites have highly distinct life cycles, pathologies, transmission strategies and surface proteomes, being dominated by the variant surface glycoprotein (African) or mucins (American) respectively. In African trypanosomes clathrin-mediated trafficking is responsible for endocytosis and post-Golgi transport, with several mechanistic aspects distinct from higher organisms. Using clathrin light chain (TcCLC) and EpsinR (TcEpsinR) as affinity handles, we identified candidate clathrin-associated proteins (CAPs) in Trypanosoma cruzi; the cohort includes orthologs of many proteins known to mediate vesicle trafficking, but significantly not the AP-2 adaptor complex. Several trypanosome-specific proteins common with African trypanosomes, were also identified. Fluorescence microscopy revealed localisations for TcEpsinR, TcCLC and TcCHC at the posterior region of trypomastigote cells, coincident with the flagellar pocket and Golgi apparatus. These data provide the first systematic analysis of clathrin-mediated trafficking in T. cruzi, allowing comparison between protein cohorts and other trypanosomes and also suggest that clathrin trafficking in at least some life stages of T. cruzi may be AP-2-independent.
authors: Laboratory of Cell Biology, Instituto Carlos Chagas/Fiocruz-PR, Rua Prof. Algacyr Munhoz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR Brazil.|Laboratory of Molecular Biology of Trypanosomes, Instituto Carlos Chagas/Fiocruz-PR, Rua Prof. Algacyr Munhoz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR Brazil.
authors: Ligia Cristina
authors: LC
authors: Kalb
authors: Laboratory of Cell Biology, Instituto Carlos Chagas/Fiocruz-PR, Rua Prof. Algacyr Munhoz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR Brazil.
authors: Yohana Camila A
authors: YC
authors: Frederico
authors: School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
authors: Cordula
authors: C
authors: Boehm
authors: Laboratory of Molecular Biology of Trypanosomes, Instituto Carlos Chagas/Fiocruz-PR, Rua Prof. Algacyr Munhoz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR Brazil.|School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
authors: Claudia Maria do Nascimento
authors: CM
authors: Moreira
authors: Laboratory of Cell Biology, Instituto Carlos Chagas/Fiocruz-PR, Rua Prof. Algacyr Munhoz Mader 3775, Cidade Industrial, 81350-010 Curitiba, PR Brazil.
authors: Maurilio José
authors: MJ
authors: Soares
authors: School of Life Sciences, University of Dundee, Dundee, DD1 5EH, UK.
authors: Mark C
authors: MC
authors: Field
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D033941:Clathrin Light Chains / Q000378:metabolism*; D022163:Clathrin-Coated Vesicles / Q000378:metabolism; D004705:Endocytosis; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D020543:Proteome; D015800:Protozoan Proteins / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000378:metabolism*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; D002966:Clathrin; D033941:Clathrin Light Chains; D020543:Proteome; D015800:Protozoan Proteins; C114154:epsin; D049452:Green Fluorescent Proteins