Endosome sorting and autophagy are essential for differentiation and virulence of Leishmania major

Sébastien Besteiro, Roderick A M Williams, Lesley S Morrison, Graham H Coombs, Jeremy C Mottram

Research output: Contribution to journalArticle

Abstract

Cellular remodeling during differentiation is essential for life-cycle progression of many unicellular eukaryotic pathogens such as Leishmania, but the mechanisms involved are largely uncharacterized. The role of endosomal sorting in differentiation was analyzed in Leishmania major by overexpression of a dominant-negative ATPase, VPS4. VPS4(E235Q) accumulated in vesicles from the endocytic pathway, and the mutant L. major was deficient in endosome sorting. Mutant parasites failed to differentiate to the obligate infective metacyclic promastigote form. Furthermore, the autophagy pathway, monitored via the expression of autophagosome marker GFP-ATG8, and shown to normally peak during initiation of metacyclogenesis, was disrupted in the mutants. The defect in late endosome-autophagosome function in the VPS4(E235Q) parasites made them less able to withstand starvation than wild-type L. major. In addition, a L. major ATG4-deficient mutant was found also to be defective in the ability to differentiate. This finding, that transformation to the infective metacyclic form is dependent on late endosome function and, more directly, autophagy, makes L. major a good model for studying the roles of these processes in differentiation.

Original languageEnglish
Pages (from-to)11384-96
Number of pages13
JournalJournal of Biological Chemistry
Volume281
Issue number16
DOIs
Publication statusPublished - 21 Apr 2006
Externally publishedYes

Fingerprint

Leishmania major
Endosomes
Autophagy
Sorting
Virulence
Pathogens
Adenosine Triphosphatases
Life cycle
Parasites
Defects
Transport Vesicles
Leishmania
Starvation
Life Cycle Stages

Keywords

  • Animals
  • Autophagy
  • Blotting, Western
  • Cell Line
  • DNA
  • DNA Primers
  • Endocytosis
  • Endosomes
  • Genes, Dominant
  • Green Fluorescent Proteins
  • Immunoblotting
  • Leishmania
  • Lysosomes
  • Macrophages
  • Microscopy, Fluorescence
  • Models, Genetic
  • Mutation
  • Plasmids
  • Protein Transport
  • Time Factors
  • Vacuoles
  • Virulence

Cite this

Besteiro, Sébastien ; Williams, Roderick A M ; Morrison, Lesley S ; Coombs, Graham H ; Mottram, Jeremy C. / Endosome sorting and autophagy are essential for differentiation and virulence of Leishmania major. In: Journal of Biological Chemistry. 2006 ; Vol. 281, No. 16. pp. 11384-96.
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Endosome sorting and autophagy are essential for differentiation and virulence of Leishmania major. / Besteiro, Sébastien; Williams, Roderick A M; Morrison, Lesley S; Coombs, Graham H; Mottram, Jeremy C.

In: Journal of Biological Chemistry, Vol. 281, No. 16, 21.04.2006, p. 11384-96.

Research output: Contribution to journalArticle

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T1 - Endosome sorting and autophagy are essential for differentiation and virulence of Leishmania major

AU - Besteiro, Sébastien

AU - Williams, Roderick A M

AU - Morrison, Lesley S

AU - Coombs, Graham H

AU - Mottram, Jeremy C

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AB - Cellular remodeling during differentiation is essential for life-cycle progression of many unicellular eukaryotic pathogens such as Leishmania, but the mechanisms involved are largely uncharacterized. The role of endosomal sorting in differentiation was analyzed in Leishmania major by overexpression of a dominant-negative ATPase, VPS4. VPS4(E235Q) accumulated in vesicles from the endocytic pathway, and the mutant L. major was deficient in endosome sorting. Mutant parasites failed to differentiate to the obligate infective metacyclic promastigote form. Furthermore, the autophagy pathway, monitored via the expression of autophagosome marker GFP-ATG8, and shown to normally peak during initiation of metacyclogenesis, was disrupted in the mutants. The defect in late endosome-autophagosome function in the VPS4(E235Q) parasites made them less able to withstand starvation than wild-type L. major. In addition, a L. major ATG4-deficient mutant was found also to be defective in the ability to differentiate. This finding, that transformation to the infective metacyclic form is dependent on late endosome function and, more directly, autophagy, makes L. major a good model for studying the roles of these processes in differentiation.

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KW - Green Fluorescent Proteins

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KW - Lysosomes

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KW - Models, Genetic

KW - Mutation

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KW - Protein Transport

KW - Time Factors

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