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Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins
Type III secretion systems (T3SS) are molecular machines in Gram-negative pathogens that translocate effector proteins with central roles in virulence. The analyses of the translocation, subcellular localization, and mode of action of T3SS effector proteins are of central importance for the understa...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6593401/ https://www.ncbi.nlm.nih.gov/pubmed/31239375 http://dx.doi.org/10.1128/mBio.00769-19 |
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author | Göser, Vera Kommnick, Carina Liss, Viktoria Hensel, Michael |
author_facet | Göser, Vera Kommnick, Carina Liss, Viktoria Hensel, Michael |
author_sort | Göser, Vera |
collection | PubMed |
description | Type III secretion systems (T3SS) are molecular machines in Gram-negative pathogens that translocate effector proteins with central roles in virulence. The analyses of the translocation, subcellular localization, and mode of action of T3SS effector proteins are of central importance for the understanding of host-pathogen interaction and pathogenesis of bacterial infections. The analysis of translocation requires dedicated techniques to address the temporal and spatial dynamics of translocation. Here we describe a novel approach to deploy self-labeling enzymes (SLE) as universal tags for localization and tracking of translocated effector proteins. Effector-SLE fusion proteins allow live-cell imaging of translocation by T3SS, superresolution microscopy, and single-molecule tracking of effector motility in living host cells. We describe the application of the approach to T3SS effector proteins for invasion and intracellular lifestyle of Salmonella enterica serovar Typhimurium and to a T3SS effector of Yersinia enterocolitica. The novel approach enables analyses of the role of T3SS in host-pathogen interaction at the highest temporal and spatial resolution, toward understanding the molecular mechanisms of their effector proteins. |
format | Online Article Text |
id | pubmed-6593401 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-65934012019-07-03 Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins Göser, Vera Kommnick, Carina Liss, Viktoria Hensel, Michael mBio Research Article Type III secretion systems (T3SS) are molecular machines in Gram-negative pathogens that translocate effector proteins with central roles in virulence. The analyses of the translocation, subcellular localization, and mode of action of T3SS effector proteins are of central importance for the understanding of host-pathogen interaction and pathogenesis of bacterial infections. The analysis of translocation requires dedicated techniques to address the temporal and spatial dynamics of translocation. Here we describe a novel approach to deploy self-labeling enzymes (SLE) as universal tags for localization and tracking of translocated effector proteins. Effector-SLE fusion proteins allow live-cell imaging of translocation by T3SS, superresolution microscopy, and single-molecule tracking of effector motility in living host cells. We describe the application of the approach to T3SS effector proteins for invasion and intracellular lifestyle of Salmonella enterica serovar Typhimurium and to a T3SS effector of Yersinia enterocolitica. The novel approach enables analyses of the role of T3SS in host-pathogen interaction at the highest temporal and spatial resolution, toward understanding the molecular mechanisms of their effector proteins. American Society for Microbiology 2019-06-25 /pmc/articles/PMC6593401/ /pubmed/31239375 http://dx.doi.org/10.1128/mBio.00769-19 Text en Copyright © 2019 Göser et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Article Göser, Vera Kommnick, Carina Liss, Viktoria Hensel, Michael Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title | Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title_full | Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title_fullStr | Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title_full_unstemmed | Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title_short | Self-Labeling Enzyme Tags for Analyses of Translocation of Type III Secretion System Effector Proteins |
title_sort | self-labeling enzyme tags for analyses of translocation of type iii secretion system effector proteins |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6593401/ https://www.ncbi.nlm.nih.gov/pubmed/31239375 http://dx.doi.org/10.1128/mBio.00769-19 |
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