Cell Stress and Chaperones

, Volume 22, Issue 5, pp 729–742 | Cite as

Hsp90 inhibitors radicicol and geldanamycin have opposing effects on Leishmania Aha1-dependent proliferation

  • Katharina Bartsch
  • Antje Hombach-Barrigah
  • Joachim ClosEmail author
Original Paper


Hsp90 and its co-chaperones are essential for the medically important parasite Leishmania donovani, facilitating life cycle control and intracellular survival. Activity of Hsp90 is regulated by co-chaperones of the Aha1 and P23 families. In this paper, we studied the expression of L. donovani Aha1 in two life cycle stages, its interaction with Hsp90 and the phenotype of Aha1 null mutants during the insect stage and inside infected macrophages. This study provides a detailed in vitro analysis of the function of Aha1 in Leishmania parasites and the first instance of a reverse genetic analysis of Aha1 in a protozoan parasite. While Aha1 is non-essential under standard growth conditions and at elevated temperature, Aha1 protects against ethanol stress. However, both overexpression and lack of Aha1 affected parasite growth in the presence of the Hsp90 inhibitors radicicol (RAD) and geldanamycin (GA). Under RAD pressure, P23 and Aha1 act in an antagonistic way. By contrast, expression levels of both co-chaperones have similar effects under GA treatment, indicating different inhibition mechanisms by the two compounds. Aha1 is also secreted in virulence-enhancing exosomes. This may explain why the loss of Aha1 reduces the infectivity of L. donovani in ex vivo mouse macrophages, indicating a role during the intracellular mammalian stage.


Leishmania Aha1 Hsp90 Co-chaperone Inhibitors Exosomes 



We thank Eugenia Bifeld and Julia Eick for providing bone marrow-derived macrophages. The work described here was funded by the Deutsche Forschungsgemeinschaft, Grant CL 120/8-1.

Supplementary material

12192_2017_800_MOESM1_ESM.pdf (62 kb)
Fig. S1 Anti-Aha1 antibody testing. (His)10: Aha1 was used to raise antibodies in a chicken. Western blot analysis showed that Aha1 antibodies recognise the natural protein in the lysates of L. major, L. donovani, the recombinant protein (His)10::Aha1, but not the orthologues in T. cruzi Y and T. cruzi Tulahuen lysates. Positions of size markers [kD] are shown on the left. (PDF 61 kb)
12192_2017_800_MOESM2_ESM.pdf (78 kb)
Fig. S2 Schematic representation of gene constructs. A. Plasmids constructed for Aha1 gene replacement. B. Schematic representation of homologous recombination between the SwaI-linearised replacement constructs and the Aha1 gene alleles. (PDF 77 kb)
12192_2017_800_MOESM3_ESM.pdf (97 kb)
Fig. S3 Cell body and flagellum length analysis of Aha1−/− null mutants and control strains. L. donovani cells of various genetic background as indicated were seeded into 10 mL of supplemented M199 (1 × 105 mL−1) and grown for 3 days at 25 °C and pH 7.0. Cells were then fixed, stained with mouse anti-α-tubulin and DAPI and visualised by fluorescence microscopy using the EVOS FL Auto epifluorescence microscope. Cell body length measurements (A) and flagellum length measurements (B) were then performed using the measuring tool of the EVOS FL Auto software. n = 2; *p ≤ 0.05,**p ≤ 0.01 (Mann-Whitney U-test) (PDF 96 kb)


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Copyright information

© Cell Stress Society International 2017

Authors and Affiliations

  1. 1.Bernhard Nocht Institute for Tropical MedicineHamburgGermany

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