The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status

Wai-Lok Yau, Pascale Pescher, Andrea MacDonald, Sonia Hem, Dorothea Zander, Silke Retzlaff, Thierry Blisnick, Brice Rotureau, Heidi Rosenqvist, Martin Wiese, Philippe Bastin, Joachim Clos, Gerald Späth

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

During its life cycle, the protozoan pathogen Leishmania donovani is exposed to contrasting environments inside insect vector and vertebrate host, to which the parasite must adapt for extra- and intracellular survival. Combining null mutant analysis with phosphorylation site-specific mutagenesis and functional complementation we genetically tested the requirement of the L. donovani chaperone cyclophilin 40 (LdCyP40) for infection. Targeted replacement of LdCyP40 had no effect on parasite viability, axenic amastigote differentiation, and resistance to various forms of environmental stress in culture, suggesting important functional redundancy to other parasite chaperones. However, ultrastructural analyses and video microscopy of cyp40-/- promastigotes uncovered important defects in cell shape, organization of the subpellicular tubulin network and motility at stationary growth phase. More importantly, cyp40-/- parasites were unable to establish intracellular infection in murine macrophages and were eliminated during the first 24 h post infection. Surprisingly, cyp40-/- infectivity was restored in complemented parasites expressing a CyP40 mutant of the unique S274 phosphorylation site. Together our data reveal non-redundant CyP40 functions in parasite cytoskeletal remodelling relevant for the development of infectious parasites in vitro independent of its phosphorylation status, and provide a framework for the genetic analysis of Leishmania-specific phosphorylation sites and their role in regulating parasite protein function.
LanguageEnglish
Pages80–97
Number of pages18
JournalMolecular Microbiology
Volume93
Issue number1
Early online date23 May 2014
DOIs
Publication statusPublished - 31 Jul 2014

Fingerprint

Leishmania donovani
Parasites
Phosphorylation
Infection
Insect Vectors
Video Microscopy
cyclophilin D
Cell Shape
Leishmania
Tubulin
Site-Directed Mutagenesis
Life Cycle Stages
Vertebrates
Macrophages

Keywords

  • Leishmania donovani
  • chaperone
  • phosphorylation
  • infection

Cite this

Yau, Wai-Lok ; Pescher, Pascale ; MacDonald, Andrea ; Hem, Sonia ; Zander, Dorothea ; Retzlaff, Silke ; Blisnick, Thierry ; Rotureau, Brice ; Rosenqvist, Heidi ; Wiese, Martin ; Bastin, Philippe ; Clos, Joachim ; Späth, Gerald. / The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status. In: Molecular Microbiology. 2014 ; Vol. 93, No. 1. pp. 80–97.
@article{7ac4fa4fca9a401daa1f6cf7add3a6db,
title = "The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status",
abstract = "During its life cycle, the protozoan pathogen Leishmania donovani is exposed to contrasting environments inside insect vector and vertebrate host, to which the parasite must adapt for extra- and intracellular survival. Combining null mutant analysis with phosphorylation site-specific mutagenesis and functional complementation we genetically tested the requirement of the L. donovani chaperone cyclophilin 40 (LdCyP40) for infection. Targeted replacement of LdCyP40 had no effect on parasite viability, axenic amastigote differentiation, and resistance to various forms of environmental stress in culture, suggesting important functional redundancy to other parasite chaperones. However, ultrastructural analyses and video microscopy of cyp40-/- promastigotes uncovered important defects in cell shape, organization of the subpellicular tubulin network and motility at stationary growth phase. More importantly, cyp40-/- parasites were unable to establish intracellular infection in murine macrophages and were eliminated during the first 24 h post infection. Surprisingly, cyp40-/- infectivity was restored in complemented parasites expressing a CyP40 mutant of the unique S274 phosphorylation site. Together our data reveal non-redundant CyP40 functions in parasite cytoskeletal remodelling relevant for the development of infectious parasites in vitro independent of its phosphorylation status, and provide a framework for the genetic analysis of Leishmania-specific phosphorylation sites and their role in regulating parasite protein function.",
keywords = "Leishmania donovani, chaperone, phosphorylation, infection",
author = "Wai-Lok Yau and Pascale Pescher and Andrea MacDonald and Sonia Hem and Dorothea Zander and Silke Retzlaff and Thierry Blisnick and Brice Rotureau and Heidi Rosenqvist and Martin Wiese and Philippe Bastin and Joachim Clos and Gerald Sp{\"a}th",
year = "2014",
month = "7",
day = "31",
doi = "10.1111/mmi.12639",
language = "English",
volume = "93",
pages = "80–97",
journal = "Molecular Microbiology",
issn = "0950-382X",
number = "1",

}

Yau, W-L, Pescher, P, MacDonald, A, Hem, S, Zander, D, Retzlaff, S, Blisnick, T, Rotureau, B, Rosenqvist, H, Wiese, M, Bastin, P, Clos, J & Späth, G 2014, 'The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status' Molecular Microbiology, vol. 93, no. 1, pp. 80–97. https://doi.org/10.1111/mmi.12639

The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status. / Yau, Wai-Lok; Pescher, Pascale; MacDonald, Andrea; Hem, Sonia; Zander, Dorothea; Retzlaff, Silke; Blisnick, Thierry; Rotureau, Brice; Rosenqvist, Heidi; Wiese, Martin; Bastin, Philippe; Clos, Joachim; Späth, Gerald.

In: Molecular Microbiology, Vol. 93, No. 1, 31.07.2014, p. 80–97.

Research output: Contribution to journalArticle

TY - JOUR

T1 - The Leishmania donovani chaperone cyclophilin 40 is essential for intracellular infection independent of its stage-specific phosphorylation status

AU - Yau, Wai-Lok

AU - Pescher, Pascale

AU - MacDonald, Andrea

AU - Hem, Sonia

AU - Zander, Dorothea

AU - Retzlaff, Silke

AU - Blisnick, Thierry

AU - Rotureau, Brice

AU - Rosenqvist, Heidi

AU - Wiese, Martin

AU - Bastin, Philippe

AU - Clos, Joachim

AU - Späth, Gerald

PY - 2014/7/31

Y1 - 2014/7/31

N2 - During its life cycle, the protozoan pathogen Leishmania donovani is exposed to contrasting environments inside insect vector and vertebrate host, to which the parasite must adapt for extra- and intracellular survival. Combining null mutant analysis with phosphorylation site-specific mutagenesis and functional complementation we genetically tested the requirement of the L. donovani chaperone cyclophilin 40 (LdCyP40) for infection. Targeted replacement of LdCyP40 had no effect on parasite viability, axenic amastigote differentiation, and resistance to various forms of environmental stress in culture, suggesting important functional redundancy to other parasite chaperones. However, ultrastructural analyses and video microscopy of cyp40-/- promastigotes uncovered important defects in cell shape, organization of the subpellicular tubulin network and motility at stationary growth phase. More importantly, cyp40-/- parasites were unable to establish intracellular infection in murine macrophages and were eliminated during the first 24 h post infection. Surprisingly, cyp40-/- infectivity was restored in complemented parasites expressing a CyP40 mutant of the unique S274 phosphorylation site. Together our data reveal non-redundant CyP40 functions in parasite cytoskeletal remodelling relevant for the development of infectious parasites in vitro independent of its phosphorylation status, and provide a framework for the genetic analysis of Leishmania-specific phosphorylation sites and their role in regulating parasite protein function.

AB - During its life cycle, the protozoan pathogen Leishmania donovani is exposed to contrasting environments inside insect vector and vertebrate host, to which the parasite must adapt for extra- and intracellular survival. Combining null mutant analysis with phosphorylation site-specific mutagenesis and functional complementation we genetically tested the requirement of the L. donovani chaperone cyclophilin 40 (LdCyP40) for infection. Targeted replacement of LdCyP40 had no effect on parasite viability, axenic amastigote differentiation, and resistance to various forms of environmental stress in culture, suggesting important functional redundancy to other parasite chaperones. However, ultrastructural analyses and video microscopy of cyp40-/- promastigotes uncovered important defects in cell shape, organization of the subpellicular tubulin network and motility at stationary growth phase. More importantly, cyp40-/- parasites were unable to establish intracellular infection in murine macrophages and were eliminated during the first 24 h post infection. Surprisingly, cyp40-/- infectivity was restored in complemented parasites expressing a CyP40 mutant of the unique S274 phosphorylation site. Together our data reveal non-redundant CyP40 functions in parasite cytoskeletal remodelling relevant for the development of infectious parasites in vitro independent of its phosphorylation status, and provide a framework for the genetic analysis of Leishmania-specific phosphorylation sites and their role in regulating parasite protein function.

KW - Leishmania donovani

KW - chaperone

KW - phosphorylation

KW - infection

UR - http://onlinelibrary.wiley.com/wol1/doi/10.1111/mmi.12639/abstract

U2 - 10.1111/mmi.12639

DO - 10.1111/mmi.12639

M3 - Article

VL - 93

SP - 80

EP - 97

JO - Molecular Microbiology

T2 - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 1

ER -