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Visualizing the Translocation and Localization of Bacterial Type III Effector Proteins by Using a Genetically Encoded Reporter System.
- Source :
-
Applied and environmental microbiology [Appl Environ Microbiol] 2016 Apr 18; Vol. 82 (9), pp. 2700-2708. Date of Electronic Publication: 2016 Apr 18 (Print Publication: 2016). - Publication Year :
- 2016
-
Abstract
- Bacterial type III secretion system (T3SS) effector proteins are critical determinants of infection for many animal and plant pathogens. However, monitoring of the translocation and delivery of these important virulence determinants has proved to be technically challenging. Here, we used a genetically engineered LOV (light-oxygen-voltage) sensing domain derivative to monitor the expression, translocation, and localization of bacterial T3SS effectors. We found the Escherichia coli O157:H7 bacterial effector fusion Tir-LOV was functional following its translocation and localized to the host cell membrane in discrete foci, demonstrating that LOV-based reporters can be used to visualize the effector translocation with minimal manipulation and interference. Further evidence for the versatility of the reporter was demonstrated by fusing LOV to the C terminus of the Shigella flexneri effector IpaB. IpaB-LOV localized preferentially at bacterial poles before translocation. We observed the rapid translocation of IpaB-LOV in a T3SS-dependent manner into host cells, where it localized at the bacterial entry site within membrane ruffles.<br /> (Copyright © 2016 Gawthorne et al.)
- Subjects :
- Bacterial Proteins genetics
Escherichia coli O157 genetics
Escherichia coli O157 metabolism
Escherichia coli Proteins genetics
Escherichia coli Proteins metabolism
Genetic Engineering methods
HeLa Cells
Host-Pathogen Interactions
Humans
Optical Imaging
Protein Domains
Shigella flexneri genetics
Shigella flexneri metabolism
Type III Secretion Systems analysis
Type III Secretion Systems genetics
Bacterial Proteins metabolism
Genes, Reporter
Type III Secretion Systems metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1098-5336
- Volume :
- 82
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Applied and environmental microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 26921426
- Full Text :
- https://doi.org/10.1128/AEM.03418-15