Cargando…

Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages

Francisella tularensis is a zoonotic intracellular pathogen that is capable of causing potentially fatal human infections. Like all successful bacterial pathogens, F. tularensis rapidly responds to changes in its environment during infection of host cells, and upon encountering different microenviro...

Descripción completa

Detalles Bibliográficos
Autores principales: Bent, Zachary W., Brazel, David M., Tran-Gyamfi, Mary B., Hamblin, Rachelle Y., VanderNoot, Victoria A., Branda, Steven S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3796476/
https://www.ncbi.nlm.nih.gov/pubmed/24155975
http://dx.doi.org/10.1371/journal.pone.0077834
_version_ 1782287485674455040
author Bent, Zachary W.
Brazel, David M.
Tran-Gyamfi, Mary B.
Hamblin, Rachelle Y.
VanderNoot, Victoria A.
Branda, Steven S.
author_facet Bent, Zachary W.
Brazel, David M.
Tran-Gyamfi, Mary B.
Hamblin, Rachelle Y.
VanderNoot, Victoria A.
Branda, Steven S.
author_sort Bent, Zachary W.
collection PubMed
description Francisella tularensis is a zoonotic intracellular pathogen that is capable of causing potentially fatal human infections. Like all successful bacterial pathogens, F. tularensis rapidly responds to changes in its environment during infection of host cells, and upon encountering different microenvironments within those cells. This ability to appropriately respond to the challenges of infection requires rapid and global shifts in gene expression patterns. In this study, we use a novel pathogen transcript enrichment strategy and whole transcriptome sequencing (RNA-Seq) to perform a detailed characterization of the rapid and global shifts in F. tularensis LVS gene expression during infection of murine macrophages. We performed differential gene expression analysis on all bacterial genes at two key stages of infection: phagosomal escape, and cytosolic replication. By comparing the F. tularensis transcriptome at these two stages of infection to that of the bacteria grown in culture, we were able to identify sets of genes that are differentially expressed over the course of infection. This analysis revealed the temporally dynamic expression of a number of known and putative transcriptional regulators and virulence factors, providing insight into their role during infection. In addition, we identified several F. tularensis genes that are significantly up-regulated during infection but had not been previously identified as virulence factors. These unknown genes may make attractive therapeutic or vaccine targets.
format Online
Article
Text
id pubmed-3796476
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-37964762013-10-23 Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages Bent, Zachary W. Brazel, David M. Tran-Gyamfi, Mary B. Hamblin, Rachelle Y. VanderNoot, Victoria A. Branda, Steven S. PLoS One Research Article Francisella tularensis is a zoonotic intracellular pathogen that is capable of causing potentially fatal human infections. Like all successful bacterial pathogens, F. tularensis rapidly responds to changes in its environment during infection of host cells, and upon encountering different microenvironments within those cells. This ability to appropriately respond to the challenges of infection requires rapid and global shifts in gene expression patterns. In this study, we use a novel pathogen transcript enrichment strategy and whole transcriptome sequencing (RNA-Seq) to perform a detailed characterization of the rapid and global shifts in F. tularensis LVS gene expression during infection of murine macrophages. We performed differential gene expression analysis on all bacterial genes at two key stages of infection: phagosomal escape, and cytosolic replication. By comparing the F. tularensis transcriptome at these two stages of infection to that of the bacteria grown in culture, we were able to identify sets of genes that are differentially expressed over the course of infection. This analysis revealed the temporally dynamic expression of a number of known and putative transcriptional regulators and virulence factors, providing insight into their role during infection. In addition, we identified several F. tularensis genes that are significantly up-regulated during infection but had not been previously identified as virulence factors. These unknown genes may make attractive therapeutic or vaccine targets. Public Library of Science 2013-10-14 /pmc/articles/PMC3796476/ /pubmed/24155975 http://dx.doi.org/10.1371/journal.pone.0077834 Text en https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Bent, Zachary W.
Brazel, David M.
Tran-Gyamfi, Mary B.
Hamblin, Rachelle Y.
VanderNoot, Victoria A.
Branda, Steven S.
Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title_full Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title_fullStr Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title_full_unstemmed Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title_short Use of a Capture-Based Pathogen Transcript Enrichment Strategy for RNA-Seq Analysis of the Francisella Tularensis LVS Transcriptome during Infection of Murine Macrophages
title_sort use of a capture-based pathogen transcript enrichment strategy for rna-seq analysis of the francisella tularensis lvs transcriptome during infection of murine macrophages
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3796476/
https://www.ncbi.nlm.nih.gov/pubmed/24155975
http://dx.doi.org/10.1371/journal.pone.0077834
work_keys_str_mv AT bentzacharyw useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages
AT brazeldavidm useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages
AT trangyamfimaryb useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages
AT hamblinrachelley useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages
AT vandernootvictoriaa useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages
AT brandastevens useofacapturebasedpathogentranscriptenrichmentstrategyforrnaseqanalysisofthefrancisellatularensislvstranscriptomeduringinfectionofmurinemacrophages