Phenotypic Characterization of a Leishmania donovani Cyclophilin 40 Null Mutant |
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| Authors: | Wai‐Lok Yau Ulrike Lambertz Lucie Colineau Pascale Pescher Andrea MacDonald Dorothea Zander Silke Retzlaff Julia Eick Neil E. Reiner Joachim Clos Gerald F. Späth |
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| Affiliation: | 1. Unité de Parasitologie Moléculaire et Signalisation, Institut Pasteur and Institut National de la Santé et de la Recherche Médicale INSERM U1201, Paris, France;2. Clos Group (Leishmanasis), Bernhard‐Nocht‐Institut für Tropenmedizin, D‐20359, Hamburg, Germany;3. Division of Infectious Diseases, Jack Bell Research Centre, University of British Columbia, Vancouver, British Columbia, Canada;4. Electron Microscopy Service, Bernhard‐Nocht‐Institut für Tropenmedizin, Hamburg, Germany |
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| Abstract: | Protozoan parasites of the genus Leishmania adapt to their arthropod and vertebrate hosts through the development of defined life cycle stages. Stage differentiation is triggered by environmental stress factors and has been linked to parasite chaperone activities. Using a null mutant approach we previously revealed important, nonredundant functions of the cochaperone cyclophilin 40 in L. donovani‐infected macrophages. Here, we characterized in more detail the virulence defect of cyp40?/? null mutants. In vitro viability assays, infection tests using macrophages, and mixed infection experiments ruled out a defect of cyp40?/? parasites in resistance to oxidative and hydrolytic stresses encountered inside the host cell phagolysosome. Investigation of the CyP40‐dependent proteome by quantitative 2D‐DiGE analysis revealed up regulation of various stress proteins in the null mutant, presumably a response to compensate for the lack of CyP40. Applying transmission electron microscopy we showed accumulation of vesicular structures in the flagellar pocket of cyp40?/? parasites that we related to a significant increase in exosome production, a phenomenon previously linked to the parasite stress response. Together these data suggest that cyp40?/? parasites experience important intrinsic homeostatic stress that likely abrogates parasite viability during intracellular infection. |
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| Keywords: | Chaperone exosomes infection null mutant analysis proteomics stress protein trypanosomatid parasite virulence |
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