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Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence
A key to the pathogenic success of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is the capacity to survive within host macrophages. Although several factors required for this survival have been identified, a comprehensive knowledge of such factors and how they work together...
Autores principales: | , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444860/ https://www.ncbi.nlm.nih.gov/pubmed/28505176 http://dx.doi.org/10.1371/journal.ppat.1006363 |
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author | Barczak, Amy K. Avraham, Roi Singh, Shantanu Luo, Samantha S. Zhang, Wei Ran Bray, Mark-Anthony Hinman, Amelia E. Thompson, Matthew Nietupski, Raymond M. Golas, Aaron Montgomery, Paul Fitzgerald, Michael Smith, Roger S. White, Dylan W. Tischler, Anna D. Carpenter, Anne E. Hung, Deborah T. |
author_facet | Barczak, Amy K. Avraham, Roi Singh, Shantanu Luo, Samantha S. Zhang, Wei Ran Bray, Mark-Anthony Hinman, Amelia E. Thompson, Matthew Nietupski, Raymond M. Golas, Aaron Montgomery, Paul Fitzgerald, Michael Smith, Roger S. White, Dylan W. Tischler, Anna D. Carpenter, Anne E. Hung, Deborah T. |
author_sort | Barczak, Amy K. |
collection | PubMed |
description | A key to the pathogenic success of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is the capacity to survive within host macrophages. Although several factors required for this survival have been identified, a comprehensive knowledge of such factors and how they work together to manipulate the host environment to benefit bacterial survival are not well understood. To systematically identify Mtb factors required for intracellular growth, we screened an arrayed, non-redundant Mtb transposon mutant library by high-content imaging to characterize the mutant-macrophage interaction. Based on a combination of imaging features, we identified mutants impaired for intracellular survival. We then characterized the phenotype of infection with each mutant by profiling the induced macrophage cytokine response. Taking a systems-level approach to understanding the biology of identified mutants, we performed a multiparametric analysis combining pathogen and host phenotypes to predict functional relationships between mutants based on clustering. Strikingly, mutants defective in two well-known virulence factors, the ESX-1 protein secretion system and the virulence lipid phthiocerol dimycocerosate (PDIM), clustered together. Building upon the shared phenotype of loss of the macrophage type I interferon (IFN) response to infection, we found that PDIM production and export are required for coordinated secretion of ESX-1-substrates, for phagosomal permeabilization, and for downstream induction of the type I IFN response. Multiparametric clustering also identified two novel genes that are required for PDIM production and induction of the type I IFN response. Thus, multiparametric analysis combining host and pathogen infection phenotypes can be used to identify novel functional relationships between genes that play a role in infection. |
format | Online Article Text |
id | pubmed-5444860 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-54448602017-06-06 Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence Barczak, Amy K. Avraham, Roi Singh, Shantanu Luo, Samantha S. Zhang, Wei Ran Bray, Mark-Anthony Hinman, Amelia E. Thompson, Matthew Nietupski, Raymond M. Golas, Aaron Montgomery, Paul Fitzgerald, Michael Smith, Roger S. White, Dylan W. Tischler, Anna D. Carpenter, Anne E. Hung, Deborah T. PLoS Pathog Research Article A key to the pathogenic success of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is the capacity to survive within host macrophages. Although several factors required for this survival have been identified, a comprehensive knowledge of such factors and how they work together to manipulate the host environment to benefit bacterial survival are not well understood. To systematically identify Mtb factors required for intracellular growth, we screened an arrayed, non-redundant Mtb transposon mutant library by high-content imaging to characterize the mutant-macrophage interaction. Based on a combination of imaging features, we identified mutants impaired for intracellular survival. We then characterized the phenotype of infection with each mutant by profiling the induced macrophage cytokine response. Taking a systems-level approach to understanding the biology of identified mutants, we performed a multiparametric analysis combining pathogen and host phenotypes to predict functional relationships between mutants based on clustering. Strikingly, mutants defective in two well-known virulence factors, the ESX-1 protein secretion system and the virulence lipid phthiocerol dimycocerosate (PDIM), clustered together. Building upon the shared phenotype of loss of the macrophage type I interferon (IFN) response to infection, we found that PDIM production and export are required for coordinated secretion of ESX-1-substrates, for phagosomal permeabilization, and for downstream induction of the type I IFN response. Multiparametric clustering also identified two novel genes that are required for PDIM production and induction of the type I IFN response. Thus, multiparametric analysis combining host and pathogen infection phenotypes can be used to identify novel functional relationships between genes that play a role in infection. Public Library of Science 2017-05-15 /pmc/articles/PMC5444860/ /pubmed/28505176 http://dx.doi.org/10.1371/journal.ppat.1006363 Text en © 2017 Barczak et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Barczak, Amy K. Avraham, Roi Singh, Shantanu Luo, Samantha S. Zhang, Wei Ran Bray, Mark-Anthony Hinman, Amelia E. Thompson, Matthew Nietupski, Raymond M. Golas, Aaron Montgomery, Paul Fitzgerald, Michael Smith, Roger S. White, Dylan W. Tischler, Anna D. Carpenter, Anne E. Hung, Deborah T. Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title | Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title_full | Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title_fullStr | Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title_full_unstemmed | Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title_short | Systematic, multiparametric analysis of Mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
title_sort | systematic, multiparametric analysis of mycobacterium tuberculosis intracellular infection offers insight into coordinated virulence |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5444860/ https://www.ncbi.nlm.nih.gov/pubmed/28505176 http://dx.doi.org/10.1371/journal.ppat.1006363 |
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