Title |
Visualizing the Translocation and Localization of Bacterial Type III Effector Proteins by Using a Genetically Encoded Reporter System
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Published in |
Applied and Environmental Microbiology, April 2016
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DOI | 10.1128/aem.03418-15 |
Pubmed ID | |
Authors |
Jayde A. Gawthorne, Laurent Audry, Claire McQuitty, Paul Dean, John M. Christie, Jost Enninga, Andrew J. Roe |
Abstract |
Bacterial Type Three 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. |
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Geographical breakdown
Country | Count | As % |
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United States | 2 | 22% |
United Kingdom | 2 | 22% |
France | 2 | 22% |
Zimbabwe | 1 | 11% |
Unknown | 2 | 22% |
Demographic breakdown
Type | Count | As % |
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Scientists | 7 | 78% |
Members of the public | 2 | 22% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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United Kingdom | 1 | 1% |
United States | 1 | 1% |
Netherlands | 1 | 1% |
France | 1 | 1% |
Unknown | 92 | 96% |
Demographic breakdown
Readers by professional status | Count | As % |
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Student > Ph. D. Student | 20 | 21% |
Researcher | 19 | 20% |
Student > Master | 13 | 14% |
Student > Bachelor | 10 | 10% |
Student > Doctoral Student | 5 | 5% |
Other | 15 | 16% |
Unknown | 14 | 15% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 37 | 39% |
Immunology and Microbiology | 19 | 20% |
Agricultural and Biological Sciences | 14 | 15% |
Chemical Engineering | 2 | 2% |
Chemistry | 2 | 2% |
Other | 6 | 6% |
Unknown | 16 | 17% |