Open AccessQuantitative Dextran Trafficking to the Coxiella burnetii Parasitophorous Vacuole
Author(s) -
Winfree Seth,
Gilk Stacey D.
Publication year - 2017
Publication title -
current protocols in microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 35
eISSN - 1934-8533
pISSN - 1934-8525
DOI - 10.1002/cpmc.34
Subject(s) - coxiella burnetii , vacuole , endosome , biology , microbiology and biotechnology , endocytic cycle , obligate , q fever , virology , cell , endocytosis , intracellular , genetics , cytoplasm , ecology
The gram‐negative bacterium Coxiella burnetii causes human Q fever, a disease characterized by a debilitating flu‐like illness in acute cases and endocarditis in chronic patients. An obligate intracellular pathogen, Coxiella burnetii survives within a large, lysosome‐like vacuole inside the host cell. A unique feature of the Coxiella parasitophorous vacuole (PV) is high levels of fusion with the host endocytic pathway, with PV‐endosome fusion critical for Coxiella survival within the host cell. This unit describes quantitating PV‐endosome fusion by measuring delivery of the fluid phase endosome marker dextran to the PV using live cell imaging. To study the effect of host cell proteins involved in PV‐endosome fusion, details are provided for using siRNA knockdown host cells. This method is a powerful tool for understanding mechanisms underlying Coxiella ’s ability to manipulate host cell trafficking pathways. © 2017 by John Wiley & Sons, Inc.