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3D correlative electron microscopy reveals continuity of Brucella-containing vacuoles with the endoplasmic reticulum.

  • Sedzicki J Focal Area Infection Biology, Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Tschon T Focal Area Infection Biology, Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Low SH Focal Area Infection Biology, Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Willemart K Microorganisms Biology Research Unit (URBM, Unité de Recherche en Biologie des Microorganismes), University of Namur, 5000 Namur, Belgium.
  • Goldie KN Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Letesson JJ Microorganisms Biology Research Unit (URBM, Unité de Recherche en Biologie des Microorganismes), University of Namur, 5000 Namur, Belgium.
  • Stahlberg H Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, 4056 Basel, Switzerland.
  • Dehio C Focal Area Infection Biology, Biozentrum, University of Basel, 4056 Basel, Switzerland christoph.dehio@unibas.ch.
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  • 2018-01-24
Published in:
  • Journal of cell science. - 2018
CLEM
Correlative light and electron microscopy
Endoplasmic reticulum
FIB/SEM tomography
Replicative Brucella-containing vacuole
Sec61
rBCV
Animals
Brucella abortus
Brucella melitensis
Cytoplasm
Endoplasmic Reticulum
HeLa Cells
Host-Pathogen Interactions
Humans
Mice
Microscopy, Electron, Scanning
Trophoblasts
Type IV Secretion Systems
Vacuoles
English Entry of the facultative intracellular pathogen Brucella into host cells results in the formation of endosomal Brucella-containing vacuoles (eBCVs) that initially traffic along the endocytic pathway. eBCV acidification triggers the expression of a type IV secretion system that translocates bacterial effector proteins into host cells. This interferes with lysosomal fusion of eBCVs and supports their maturation to replicative Brucella-containing vacuoles (rBCVs). Bacteria replicate in rBCVs to large numbers, eventually occupying most of the cytoplasmic volume. As rBCV membranes tightly wrap each individual bacterium, they are constantly being expanded and remodeled during exponential bacterial growth. rBCVs are known to carry endoplasmic reticulum (ER) markers; however, the relationship of the vacuole to the genuine ER has remained elusive. Here, we have reconstructed the 3-dimensional ultrastructure of rBCVs and associated ER by correlative structured illumination microscopy (SIM) and focused ion beam/scanning electron microscopic tomography (FIB/SEM). Studying B. abortus-infected HeLa cells and trophoblasts derived from B. melitensis-infected mice, we demonstrate that rBCVs are complex and interconnected compartments that are continuous with neighboring ER cisternae, thus supporting a model that rBCVs are extensions of genuine ER.
Language
  • English
Open access status
bronze
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Persistent URL
https://sonar.ch/global/documents/17720
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