Gawthorne, J. A., Audry, L., McQuitty, C., Dean, P., Christie, J. M. , Enninga, J. and Roe, A. J. (2016) Visualizing translocation and localization of bacterial type III effector proteins using a genetically encoded reporter system. Applied and Environmental Microbiology, 82(9), pp. 2700-2708. (doi: 10.1128/AEM.03418-15) (PMID:25576618) (PMCID:PMC4345375)
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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.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Roe, Professor Andrew and Gawthorne, Ms Jayde and Christie, Professor John |
| Authors: | Gawthorne, J. A., Audry, L., McQuitty, C., Dean, P., Christie, J. M., Enninga, J., and Roe, A. J. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity College of Medical Veterinary and Life Sciences > School of Molecular Biosciences |
| Journal Name: | Applied and Environmental Microbiology |
| Publisher: | American Society for Microbiology |
| ISSN: | 0099-2240 |
| ISSN (Online): | 1098-5336 |
| Published Online: | 26 February 2016 |
| Copyright Holders: | Copyright © 2016 The Authors |
| First Published: | First published in Applied and Environmental Microbiology 82(9):2700-2708 |
| Publisher Policy: | Reproduced under a Creative Commons License |
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