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

Jaroslaw Sedzicki, Therese Tschon, Shyan Huey Low, Kevin Willemart, Kenneth N Goldie, Jean-Jacques Letesson, Henning Stahlberg, Christoph Dehio

Research output: Contribution to journalArticle

Abstract

Entry of the facultative intracellular pathogenBrucellainto host cells results in the formation of endosomalBrucella-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 replicativeBrucella-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). StudyingB. abortus-infected HeLa cells and trophoblasts derived fromB. 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.

LanguageEnglish
JournalJournal of Cell Science
Volume131
Issue number4
DOIs
Publication statusPublished - 1 Feb 2018
Externally publishedYes

Fingerprint

Vacuoles
Endoplasmic Reticulum
Electron Microscopy
Electron Microscope Tomography
Bacteria
Bacterial Proteins
Trophoblasts
Lighting
HeLa Cells
Microscopy
Ions
Membranes

Keywords

  • CLEM
  • Correlative light and electron microscopy
  • Endoplasmic reticulum
  • FIB/SEM tomography
  • RBCV
  • Replicative Brucellacontaining vacuole
  • Sec61

Cite this

Sedzicki, Jaroslaw ; Tschon, Therese ; Low, Shyan Huey ; Willemart, Kevin ; Goldie, Kenneth N ; Letesson, Jean-Jacques ; Stahlberg, Henning ; Dehio, Christoph. / 3D correlative electron microscopy reveals continuity of-containing vacuoles with the endoplasmic reticulum. In: Journal of Cell Science. 2018 ; Vol. 131, No. 4.
@article{d1a0591d8e43495c8e0ab481c7df8b1b,
title = "3D correlative electron microscopy reveals continuity of-containing vacuoles with the endoplasmic reticulum",
abstract = "Entry of the facultative intracellular pathogenBrucellainto host cells results in the formation of endosomalBrucella-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 replicativeBrucella-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). StudyingB. abortus-infected HeLa cells and trophoblasts derived fromB. 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.",
keywords = "CLEM, Correlative light and electron microscopy, Endoplasmic reticulum, FIB/SEM tomography, RBCV, Replicative Brucellacontaining vacuole, Sec61",
author = "Jaroslaw Sedzicki and Therese Tschon and Low, {Shyan Huey} and Kevin Willemart and Goldie, {Kenneth N} and Jean-Jacques Letesson and Henning Stahlberg and Christoph Dehio",
note = "{\circledC} 2018. Published by The Company of Biologists Ltd.",
year = "2018",
month = "2",
day = "1",
doi = "10.1242/jcs.210799",
language = "English",
volume = "131",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "The Company of Biologists",
number = "4",

}

3D correlative electron microscopy reveals continuity of-containing vacuoles with the endoplasmic reticulum. / Sedzicki, Jaroslaw; Tschon, Therese; Low, Shyan Huey; Willemart, Kevin; Goldie, Kenneth N; Letesson, Jean-Jacques; Stahlberg, Henning; Dehio, Christoph.

In: Journal of Cell Science, Vol. 131, No. 4, 01.02.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - 3D correlative electron microscopy reveals continuity of-containing vacuoles with the endoplasmic reticulum

AU - Sedzicki, Jaroslaw

AU - Tschon, Therese

AU - Low, Shyan Huey

AU - Willemart, Kevin

AU - Goldie, Kenneth N

AU - Letesson, Jean-Jacques

AU - Stahlberg, Henning

AU - Dehio, Christoph

N1 - © 2018. Published by The Company of Biologists Ltd.

PY - 2018/2/1

Y1 - 2018/2/1

N2 - Entry of the facultative intracellular pathogenBrucellainto host cells results in the formation of endosomalBrucella-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 replicativeBrucella-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). StudyingB. abortus-infected HeLa cells and trophoblasts derived fromB. 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.

AB - Entry of the facultative intracellular pathogenBrucellainto host cells results in the formation of endosomalBrucella-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 replicativeBrucella-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). StudyingB. abortus-infected HeLa cells and trophoblasts derived fromB. 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.

KW - CLEM

KW - Correlative light and electron microscopy

KW - Endoplasmic reticulum

KW - FIB/SEM tomography

KW - RBCV

KW - Replicative Brucellacontaining vacuole

KW - Sec61

UR - http://www.scopus.com/inward/record.url?scp=85042395629&partnerID=8YFLogxK

U2 - 10.1242/jcs.210799

DO - 10.1242/jcs.210799

M3 - Article

VL - 131

JO - Journal of Cell Science

T2 - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 4

ER -