1. Phenotypic Characterization of a Leishmania donovani Cyclophilin 40 Null Mutant
- Author
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Wai-Lok Yau, Gerald F. Späth, Neil E. Reiner, Joachim Clos, Dorothea Zander, Andrea MacDonald, Silke Retzlaff, Julia Eick, Ulrike Lambertz, Lucie Colineau, Pascale Pescher, Parasitologie moléculaire et Signalisation / Molecular Parasitology and Signaling, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Clos Group (Leishmanasis), Bernhard Nocht Institute for Tropical Medicine - Bernhard-Nocht-Institut für Tropenmedizin [Hamburg, Germany] (BNITM), Division of Infectious Disease [Vancouver], University of British Columbia (UBC)-Jack Bell Research Centre [Vancouver], Electron Microscopy Service, 7th Framework Programme of the European Commission. Grant Number: 223414French Government's Investissements d'Avenir programme: Laboratoire d'Excellence ‘Integrative Biology of Emerging Infectious Diseases’. Grant Number: ANR-10-LABX-62-IBEIDFondation pour la Recherche Médicale. Grant Number: FDT20100920296Leibniz-DAAD programmeCanadian Institutes of Health Research. Grant Number: MOP# 125879, ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), European Project: 223414,EC:FP7:HEALTH,FP7-HEALTH-2007-B,LEISHDRUG(2008), Ricard Andraos, Christel, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Targeting the Leishmania kinome for the development of novel anti-parasitic strategies - LEISHDRUG - - EC:FP7:HEALTH2008-10-01 - 2012-03-31 - 223414 - VALID, and Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)
- Subjects
0301 basic medicine ,trypanosomatid parasite ,Mutant ,null mutant analysis ,Leishmania donovani ,Protozoan Proteins ,Virulence ,Chaperone ,exosomes ,Biology ,Microbiology ,Phagolysosome ,03 medical and health sciences ,Cyclophilins ,Mice ,proteomics ,Heat shock protein ,Animals ,Humans ,Electrophoresis, Gel, Two-Dimensional ,[SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology ,Cyclophilin ,Heat-Shock Proteins ,Macrophages ,biology.organism_classification ,infection ,Cell biology ,virulence ,Mice, Inbred C57BL ,030104 developmental biology ,[SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology ,Phenotype ,Proteome ,Mutation ,Leishmaniasis, Visceral ,Intracellular ,stress protein ,Cyclophilin D - Abstract
International audience; 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.
- Published
- 2016
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