Format

Send to

Choose Destination
J Vis Exp. 2014 Sep 9;(91):e51601. doi: 10.3791/51601.

Use of Shigella flexneri to study autophagy-cytoskeleton interactions.

Author information

1
Section of Microbiology, MRC Centre for Molecular Bacteriology and Infection, Imperial College London.
2
Département de Biologie du Développement et des Cellules Souches, Institut Pasteur, Unité Macrophages et Développement de l'Immunité
3
Section of Microbiology, MRC Centre for Molecular Bacteriology and Infection, Imperial College London; s.mostowy@imperial.ac.uk.

Abstract

Shigella flexneri is an intracellular pathogen that can escape from phagosomes to reach the cytosol, and polymerize the host actin cytoskeleton to promote its motility and dissemination. New work has shown that proteins involved in actin-based motility are also linked to autophagy, an intracellular degradation process crucial for cell autonomous immunity. Strikingly, host cells may prevent actin-based motility of S. flexneri by compartmentalizing bacteria inside 'septin cages' and targeting them to autophagy. These observations indicate that a more complete understanding of septins, a family of filamentous GTP-binding proteins, will provide new insights into the process of autophagy. This report describes protocols to monitor autophagy-cytoskeleton interactions caused by S. flexneri in vitro using tissue culture cells and in vivo using zebrafish larvae. These protocols enable investigation of intracellular mechanisms that control bacterial dissemination at the molecular, cellular, and whole organism level.

PMID:
25226510
PMCID:
PMC4823020
DOI:
10.3791/51601
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for MyJove Corporation Icon for PubMed Central
Loading ...
Support Center