The Autophagic Pathway Is Actively Modulated by Phase II Coxiella Burnetii to Efficiently Replicate in the Host Cell

Cell Microbiol. 2007 Apr;9(4):891-909. doi: 10.1111/j.1462-5822.2006.00838.x. Epub 2006 Nov 3.

Abstract

The etiologic agent of Q fever Coxiella burnetii, is an intracellular obligate parasite that develops large vacuoles with phagolysosomal characteristics, containing multiple replicating bacteria. We have previously shown that Phase II C. burnetii replicative vacuoles generated after 24-48 h post infection are decorated with the autophagic protein LC3. The aim of the present study was to examine, at earlier stages of infection, the distribution and roles of the small GTPases Rab5 and Rab7, markers of early and late endosomes respectively, as well as of the protein LC3 on C. burnetii trafficking. Our results indicate that: (i) Coxiella phagosomes (Cph) acquire the two Rab proteins sequentially during infection; (ii) overexpression of a dominant negative mutant form of Rab5, but not of Rab7, impaired Coxiella entry, whereas both Rab5 and Rab7 dominant negative mutants inhibited vacuole formation; (iii) Cph colocalized with the protein LC3 as early as 5 min after infection; acquisition of this protein appeared to be a bacterially driven process, because it was inhibited by the bacteriostatic antibiotic chloramphenicol and (iv) C. burnetii delayed the arrival of the typical lysosomal protease cathepsin D to the Cph, which delay is further increased by starvation-induced autophagy. Based on our results we propose that C. burnetii transits through the normal endo/phagocytic pathway but actively interacts with autophagosomes at early times after infection. This intersection with the autophagic pathway delays fusion with the lysosomal compartment possibly favouring the intracellular differentiation and survival of the bacteria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autophagy / genetics
  • Autophagy / physiology*
  • Blotting, Western
  • CHO Cells
  • Cathepsins / genetics
  • Cathepsins / metabolism
  • Coxiella burnetii / growth & development*
  • Cricetinae
  • Cricetulus
  • Fluorescent Antibody Technique, Indirect
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lysosomes / metabolism
  • Lysosomes / microbiology
  • Microscopy, Fluorescence
  • Phagosomes / metabolism
  • Signal Transduction / genetics
  • Signal Transduction / physiology*
  • Vacuoles / metabolism
  • Vacuoles / microbiology
  • rab GTP-Binding Proteins / genetics
  • rab GTP-Binding Proteins / metabolism
  • rab5 GTP-Binding Proteins / genetics
  • rab5 GTP-Binding Proteins / metabolism

Substances

  • Green Fluorescent Proteins
  • rab7 protein
  • Cathepsins
  • rab GTP-Binding Proteins
  • rab5 GTP-Binding Proteins