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Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network

The human pathogen Coxiella burnetii causes Q-fever and is classified as a category B bio-weapon. Exploiting the development of the axenic growth medium ACCM-2, we have now used (13)C-labeling experiments and isotopolog profiling to investigate the highly diverse metabolic network of C. burnetii. To...

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Autores principales: Häuslein, Ina, Cantet, Franck, Reschke, Sarah, Chen, Fan, Bonazzi, Matteo, Eisenreich, Wolfgang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5489692/
https://www.ncbi.nlm.nih.gov/pubmed/28706879
http://dx.doi.org/10.3389/fcimb.2017.00285
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author Häuslein, Ina
Cantet, Franck
Reschke, Sarah
Chen, Fan
Bonazzi, Matteo
Eisenreich, Wolfgang
author_facet Häuslein, Ina
Cantet, Franck
Reschke, Sarah
Chen, Fan
Bonazzi, Matteo
Eisenreich, Wolfgang
author_sort Häuslein, Ina
collection PubMed
description The human pathogen Coxiella burnetii causes Q-fever and is classified as a category B bio-weapon. Exploiting the development of the axenic growth medium ACCM-2, we have now used (13)C-labeling experiments and isotopolog profiling to investigate the highly diverse metabolic network of C. burnetii. To this aim, C. burnetii RSA 439 NMII was cultured in ACCM-2 containing 5 mM of either [U-(13)C(3)]serine, [U-(13)C(6)]glucose, or [U-(13)C(3)]glycerol until the late-logarithmic phase. GC/MS-based isotopolog profiling of protein-derived amino acids, methanol-soluble polar metabolites, fatty acids, and cell wall components (e.g., diaminopimelate and sugars) from the labeled bacteria revealed differential incorporation rates and isotopolog profiles. These data served to decipher the diverse usages of the labeled substrates and the relative carbon fluxes into the core metabolism of the pathogen. Whereas, de novo biosynthesis from any of these substrates could not be found for histidine, isoleucine, leucine, lysine, phenylalanine, proline and valine, the other amino acids and metabolites under study acquired (13)C-label at specific rates depending on the nature of the tracer compound. Glucose was directly used for cell wall biosynthesis, but was also converted into pyruvate (and its downstream metabolites) through the glycolytic pathway or into erythrose 4-phosphate (e.g., for the biosynthesis of tyrosine) via the non-oxidative pentose phosphate pathway. Glycerol efficiently served as a gluconeogenetic substrate and could also be used via phosphoenolpyruvate and diaminopimelate as a major carbon source for cell wall biosynthesis. In contrast, exogenous serine was mainly utilized in downstream metabolic processes, e.g., via acetyl-CoA in a complete citrate cycle with fluxes in the oxidative direction and as a carbon feed for fatty acid biosynthesis. In summary, the data reflect multiple and differential substrate usages by C. burnetii in a bipartite-type metabolic network, resembling the overall topology of the related pathogen Legionella pneumophila. These strategies could benefit the metabolic capacities of the pathogens also as a trait to adapt for replication under intracellular conditions.
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spelling pubmed-54896922017-07-13 Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network Häuslein, Ina Cantet, Franck Reschke, Sarah Chen, Fan Bonazzi, Matteo Eisenreich, Wolfgang Front Cell Infect Microbiol Microbiology The human pathogen Coxiella burnetii causes Q-fever and is classified as a category B bio-weapon. Exploiting the development of the axenic growth medium ACCM-2, we have now used (13)C-labeling experiments and isotopolog profiling to investigate the highly diverse metabolic network of C. burnetii. To this aim, C. burnetii RSA 439 NMII was cultured in ACCM-2 containing 5 mM of either [U-(13)C(3)]serine, [U-(13)C(6)]glucose, or [U-(13)C(3)]glycerol until the late-logarithmic phase. GC/MS-based isotopolog profiling of protein-derived amino acids, methanol-soluble polar metabolites, fatty acids, and cell wall components (e.g., diaminopimelate and sugars) from the labeled bacteria revealed differential incorporation rates and isotopolog profiles. These data served to decipher the diverse usages of the labeled substrates and the relative carbon fluxes into the core metabolism of the pathogen. Whereas, de novo biosynthesis from any of these substrates could not be found for histidine, isoleucine, leucine, lysine, phenylalanine, proline and valine, the other amino acids and metabolites under study acquired (13)C-label at specific rates depending on the nature of the tracer compound. Glucose was directly used for cell wall biosynthesis, but was also converted into pyruvate (and its downstream metabolites) through the glycolytic pathway or into erythrose 4-phosphate (e.g., for the biosynthesis of tyrosine) via the non-oxidative pentose phosphate pathway. Glycerol efficiently served as a gluconeogenetic substrate and could also be used via phosphoenolpyruvate and diaminopimelate as a major carbon source for cell wall biosynthesis. In contrast, exogenous serine was mainly utilized in downstream metabolic processes, e.g., via acetyl-CoA in a complete citrate cycle with fluxes in the oxidative direction and as a carbon feed for fatty acid biosynthesis. In summary, the data reflect multiple and differential substrate usages by C. burnetii in a bipartite-type metabolic network, resembling the overall topology of the related pathogen Legionella pneumophila. These strategies could benefit the metabolic capacities of the pathogens also as a trait to adapt for replication under intracellular conditions. Frontiers Media S.A. 2017-06-29 /pmc/articles/PMC5489692/ /pubmed/28706879 http://dx.doi.org/10.3389/fcimb.2017.00285 Text en Copyright © 2017 Häuslein, Cantet, Reschke, Chen, Bonazzi and Eisenreich. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Häuslein, Ina
Cantet, Franck
Reschke, Sarah
Chen, Fan
Bonazzi, Matteo
Eisenreich, Wolfgang
Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title_full Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title_fullStr Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title_full_unstemmed Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title_short Multiple Substrate Usage of Coxiella burnetii to Feed a Bipartite Metabolic Network
title_sort multiple substrate usage of coxiella burnetii to feed a bipartite metabolic network
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5489692/
https://www.ncbi.nlm.nih.gov/pubmed/28706879
http://dx.doi.org/10.3389/fcimb.2017.00285
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