Cargando…
Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library
The human-adapted Salmonella enterica serovar Typhi (S. Typhi) causes a systemic infection known as typhoid fever. This disease relies on the ability of the bacterium to survive within macrophages. In order to identify genes involved during interaction with macrophages, a pool of approximately 10(5)...
Autores principales: | , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3344905/ https://www.ncbi.nlm.nih.gov/pubmed/22574205 http://dx.doi.org/10.1371/journal.pone.0036643 |
_version_ | 1782232095744065536 |
---|---|
author | Sabbagh, Sébastien C. Lepage, Christine McClelland, Michael Daigle, France |
author_facet | Sabbagh, Sébastien C. Lepage, Christine McClelland, Michael Daigle, France |
author_sort | Sabbagh, Sébastien C. |
collection | PubMed |
description | The human-adapted Salmonella enterica serovar Typhi (S. Typhi) causes a systemic infection known as typhoid fever. This disease relies on the ability of the bacterium to survive within macrophages. In order to identify genes involved during interaction with macrophages, a pool of approximately 10(5) transposon mutants of S. Typhi was subjected to three serial passages of 24 hours through human macrophages. Mutants recovered from infected macrophages (output) were compared to the initial pool (input) and those significantly underrepresented resulted in the identification of 130 genes encoding for cell membrane components, fimbriae, flagella, regulatory processes, pathogenesis, and many genes of unknown function. Defined deletions in 28 genes or gene clusters were created and mutants were evaluated in competitive and individual infection assays for uptake and intracellular survival during interaction with human macrophages. Overall, 26 mutants had defects in the competitive assay and 14 mutants had defects in the individual assay. Twelve mutants had defects in both assays, including acrA, exbDB, flhCD, fliC, gppA, mlc, pgtE, typA, waaQGP, SPI-4, STY1867-68, and STY2346. The complementation of several mutants by expression of plasmid-borne wild-type genes or gene clusters reversed defects, confirming that the phenotypic impairments within macrophages were gene-specific. In this study, 35 novel phenotypes of either uptake or intracellular survival in macrophages were associated with Salmonella genes. Moreover, these results reveal several genes encoding molecular mechanisms not previously known to be involved in systemic infection by human-adapted typhoidal Salmonella that will need to be elucidated. |
format | Online Article Text |
id | pubmed-3344905 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33449052012-05-09 Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library Sabbagh, Sébastien C. Lepage, Christine McClelland, Michael Daigle, France PLoS One Research Article The human-adapted Salmonella enterica serovar Typhi (S. Typhi) causes a systemic infection known as typhoid fever. This disease relies on the ability of the bacterium to survive within macrophages. In order to identify genes involved during interaction with macrophages, a pool of approximately 10(5) transposon mutants of S. Typhi was subjected to three serial passages of 24 hours through human macrophages. Mutants recovered from infected macrophages (output) were compared to the initial pool (input) and those significantly underrepresented resulted in the identification of 130 genes encoding for cell membrane components, fimbriae, flagella, regulatory processes, pathogenesis, and many genes of unknown function. Defined deletions in 28 genes or gene clusters were created and mutants were evaluated in competitive and individual infection assays for uptake and intracellular survival during interaction with human macrophages. Overall, 26 mutants had defects in the competitive assay and 14 mutants had defects in the individual assay. Twelve mutants had defects in both assays, including acrA, exbDB, flhCD, fliC, gppA, mlc, pgtE, typA, waaQGP, SPI-4, STY1867-68, and STY2346. The complementation of several mutants by expression of plasmid-borne wild-type genes or gene clusters reversed defects, confirming that the phenotypic impairments within macrophages were gene-specific. In this study, 35 novel phenotypes of either uptake or intracellular survival in macrophages were associated with Salmonella genes. Moreover, these results reveal several genes encoding molecular mechanisms not previously known to be involved in systemic infection by human-adapted typhoidal Salmonella that will need to be elucidated. Public Library of Science 2012-05-04 /pmc/articles/PMC3344905/ /pubmed/22574205 http://dx.doi.org/10.1371/journal.pone.0036643 Text en Sabbagh 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Sabbagh, Sébastien C. Lepage, Christine McClelland, Michael Daigle, France Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title | Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title_full | Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title_fullStr | Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title_full_unstemmed | Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title_short | Selection of Salmonella enterica Serovar Typhi Genes Involved during Interaction with Human Macrophages by Screening of a Transposon Mutant Library |
title_sort | selection of salmonella enterica serovar typhi genes involved during interaction with human macrophages by screening of a transposon mutant library |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3344905/ https://www.ncbi.nlm.nih.gov/pubmed/22574205 http://dx.doi.org/10.1371/journal.pone.0036643 |
work_keys_str_mv | AT sabbaghsebastienc selectionofsalmonellaentericaserovartyphigenesinvolvedduringinteractionwithhumanmacrophagesbyscreeningofatransposonmutantlibrary AT lepagechristine selectionofsalmonellaentericaserovartyphigenesinvolvedduringinteractionwithhumanmacrophagesbyscreeningofatransposonmutantlibrary AT mcclellandmichael selectionofsalmonellaentericaserovartyphigenesinvolvedduringinteractionwithhumanmacrophagesbyscreeningofatransposonmutantlibrary AT daiglefrance selectionofsalmonellaentericaserovartyphigenesinvolvedduringinteractionwithhumanmacrophagesbyscreeningofatransposonmutantlibrary |