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Phenotypic Characterization of a Leishmania donovani Cyclophilin 40 Null Mutant
Author(s) -
Yau WaiLok,
Lambertz Ulrike,
Colineau Lucie,
Pescher Pascale,
MacDonald Andrea,
Zander Dorothea,
Retzlaff Silke,
Eick Julia,
Reiner Neil E.,
Clos Joachim,
Späth Gerald F.
Publication year - 2016
Publication title -
journal of eukaryotic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.067
H-Index - 77
eISSN - 1550-7408
pISSN - 1066-5234
DOI - 10.1111/jeu.12329
Subject(s) - biology , microbiology and biotechnology , mutant , leishmania donovani , parasite hosting , intracellular parasite , amastigote , intracellular , leishmania , genetics , gene , leishmaniasis , visceral leishmaniasis , world wide web , computer science
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‐Di GE 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.