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Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling

BACKGROUND: The human pathogen Listeria monocytogenes resides and proliferates within the cytoplasm of epithelial cells. While the virulence factors essentially contributing to this step of the infection cycle are well characterized, the set of listerial genes contributing to intracellular replicati...

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Autores principales: Schauer, Kristina, Geginat, Gernot, Liang, Chunguang, Goebel, Werner, Dandekar, Thomas, Fuchs, Thilo M
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3091722/
https://www.ncbi.nlm.nih.gov/pubmed/20955543
http://dx.doi.org/10.1186/1471-2164-11-573
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author Schauer, Kristina
Geginat, Gernot
Liang, Chunguang
Goebel, Werner
Dandekar, Thomas
Fuchs, Thilo M
author_facet Schauer, Kristina
Geginat, Gernot
Liang, Chunguang
Goebel, Werner
Dandekar, Thomas
Fuchs, Thilo M
author_sort Schauer, Kristina
collection PubMed
description BACKGROUND: The human pathogen Listeria monocytogenes resides and proliferates within the cytoplasm of epithelial cells. While the virulence factors essentially contributing to this step of the infection cycle are well characterized, the set of listerial genes contributing to intracellular replication remains to be defined on a genome-wide level. RESULTS: A comprehensive library of L. monocytogenes strain EGD knockout mutants was constructed upon insertion-duplication mutagenesis, and 1491 mutants were tested for their phenotypes in rich medium and in a Caco-2 cell culture assay. Following sequencing of the plasmid insertion site, 141 different genes required for invasion of and replication in Caco-2 cells were identified. Ten in-frame deletion mutants were constructed that confirmed the data. The genes with known functions are mainly involved in cellular processes including transport, in the intermediary metabolism of sugars, nucleotides and lipids, and in information pathways such as regulatory functions. No function could be ascribed to 18 genes, and a counterpart of eight genes is missing in the apathogenic species L. innocua. Mice infection studies revealed the in vivo requirement of IspE (Lmo0190) involved in mevalonate synthesis, and of the novel ABC transporter Lmo0135-0137 associated with cysteine transport. Based on the data of this genome-scale screening, an extreme pathway and elementary mode analysis was applied that demonstrates the critical role of glycerol and purine metabolism, of fucose utilization, and of the synthesis of glutathione, aspartate semialdehyde, serine and branched chain amino acids during intracellular replication of L. monocytogenes. CONCLUSION: The combination of a genetic screening and a modelling approach revealed that a series of transporters help L. monocytogenes to overcome a putative lack of nutrients within cells, and that a high metabolic flexibility contributes to the intracellular replication of this pathogen.
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spelling pubmed-30917222011-05-11 Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling Schauer, Kristina Geginat, Gernot Liang, Chunguang Goebel, Werner Dandekar, Thomas Fuchs, Thilo M BMC Genomics Research Article BACKGROUND: The human pathogen Listeria monocytogenes resides and proliferates within the cytoplasm of epithelial cells. While the virulence factors essentially contributing to this step of the infection cycle are well characterized, the set of listerial genes contributing to intracellular replication remains to be defined on a genome-wide level. RESULTS: A comprehensive library of L. monocytogenes strain EGD knockout mutants was constructed upon insertion-duplication mutagenesis, and 1491 mutants were tested for their phenotypes in rich medium and in a Caco-2 cell culture assay. Following sequencing of the plasmid insertion site, 141 different genes required for invasion of and replication in Caco-2 cells were identified. Ten in-frame deletion mutants were constructed that confirmed the data. The genes with known functions are mainly involved in cellular processes including transport, in the intermediary metabolism of sugars, nucleotides and lipids, and in information pathways such as regulatory functions. No function could be ascribed to 18 genes, and a counterpart of eight genes is missing in the apathogenic species L. innocua. Mice infection studies revealed the in vivo requirement of IspE (Lmo0190) involved in mevalonate synthesis, and of the novel ABC transporter Lmo0135-0137 associated with cysteine transport. Based on the data of this genome-scale screening, an extreme pathway and elementary mode analysis was applied that demonstrates the critical role of glycerol and purine metabolism, of fucose utilization, and of the synthesis of glutathione, aspartate semialdehyde, serine and branched chain amino acids during intracellular replication of L. monocytogenes. CONCLUSION: The combination of a genetic screening and a modelling approach revealed that a series of transporters help L. monocytogenes to overcome a putative lack of nutrients within cells, and that a high metabolic flexibility contributes to the intracellular replication of this pathogen. BioMed Central 2010-10-18 /pmc/articles/PMC3091722/ /pubmed/20955543 http://dx.doi.org/10.1186/1471-2164-11-573 Text en Copyright ©2010 Schauer et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Schauer, Kristina
Geginat, Gernot
Liang, Chunguang
Goebel, Werner
Dandekar, Thomas
Fuchs, Thilo M
Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title_full Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title_fullStr Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title_full_unstemmed Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title_short Deciphering the intracellular metabolism of Listeria monocytogenes by mutant screening and modelling
title_sort deciphering the intracellular metabolism of listeria monocytogenes by mutant screening and modelling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3091722/
https://www.ncbi.nlm.nih.gov/pubmed/20955543
http://dx.doi.org/10.1186/1471-2164-11-573
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