Brucella abortus depends on pyruvate phosphate dikinase and malic enzyme but not on Fbp and GlpX fructose-1,6-bisphosphatases for full virulence in laboratory models

Amaia Zúñiga-Ripa, Thibault Barbier, Raquel Conde-Álvarez, Estrella Martínez-Gómez, Leyre Palacios-Chaves, Yolanda Gil-Ramírez, María Jesús Grilló, Jean-Jacques Letesson, Maite Iriarte, Ignacio Moriyón

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Résumé

The brucellae are the etiological agents of brucellosis, a worldwide-distributed zoonosis. These bacteria are facultative intracellular parasites, and thus are able to adjust their metabolism to the extra and intracellular environments encountered during an infectious cycle. However, this aspect of Brucella biology is imperfectly known and the nutrients available in the intracellular niche are unknown. Here, we investigated the central pathways of C metabolism used by Brucella abortus by deleting the putative fructose-1,6-bisphosphatases (fbp and glpX), phosphoenolpyruvate carboxykinase (pckA), pyruvate phosphate dikinase (ppdK) and malic enzyme (mae) genes. In gluconeogenic but not in rich media, growth of mutants ΔppdK and Δmae was severely impaired and growth of the double Δfbp-ΔglpX mutant was reduced. In macrophages, only ΔppdK and Δmae showed reduced multiplication, and studies with ΔppdK confirmed that it reached the replicative niche. Similarly, only ΔppdK and Δmae were attenuated in mice, the former being cleared by week 10 and the latter persisting longer than 12 weeks. We also investigated the glyoxylate cycle. Although aceA (isocitrate lyase) promoter activity was enhanced in rich medium, aceA disruption had no effect in vitro or on multiplication in macrophages or mouse spleens. The results suggest that B. abortus grows intracellularly using a limited supply of 6 C (and 5 C) sugars that is compensated by glutamate and possibly other amino acids entering the Krebs cycle without a critical role of the glyoxylate shunt.

langue originaleAnglais
journalJournal of Bacteriology
Les DOIs
étatPublié - 2014

Empreinte digitale

Orthophosphate Dikinase Pyruvate
Brucella abortus
Fructose
Virulence
Enzymes
Brucella
Macrophages
Isocitrate Lyase
Phosphoenolpyruvate
Citric Acid Cycle
Brucellosis
Zoonoses
Growth
Glutamic Acid
Parasites
Spleen
Bacteria
Amino Acids
Food
Genes

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Zúñiga-Ripa, Amaia ; Barbier, Thibault ; Conde-Álvarez, Raquel ; Martínez-Gómez, Estrella ; Palacios-Chaves, Leyre ; Gil-Ramírez, Yolanda ; Grilló, María Jesús ; Letesson, Jean-Jacques ; Iriarte, Maite ; Moriyón, Ignacio. / Brucella abortus depends on pyruvate phosphate dikinase and malic enzyme but not on Fbp and GlpX fructose-1,6-bisphosphatases for full virulence in laboratory models. Dans: Journal of Bacteriology. 2014.
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title = "Brucella abortus depends on pyruvate phosphate dikinase and malic enzyme but not on Fbp and GlpX fructose-1,6-bisphosphatases for full virulence in laboratory models",
abstract = "The brucellae are the etiological agents of brucellosis, a worldwide-distributed zoonosis. These bacteria are facultative intracellular parasites, and thus are able to adjust their metabolism to the extra and intracellular environments encountered during an infectious cycle. However, this aspect of Brucella biology is imperfectly known and the nutrients available in the intracellular niche are unknown. Here, we investigated the central pathways of C metabolism used by Brucella abortus by deleting the putative fructose-1,6-bisphosphatases (fbp and glpX), phosphoenolpyruvate carboxykinase (pckA), pyruvate phosphate dikinase (ppdK) and malic enzyme (mae) genes. In gluconeogenic but not in rich media, growth of mutants ΔppdK and Δmae was severely impaired and growth of the double Δfbp-ΔglpX mutant was reduced. In macrophages, only ΔppdK and Δmae showed reduced multiplication, and studies with ΔppdK confirmed that it reached the replicative niche. Similarly, only ΔppdK and Δmae were attenuated in mice, the former being cleared by week 10 and the latter persisting longer than 12 weeks. We also investigated the glyoxylate cycle. Although aceA (isocitrate lyase) promoter activity was enhanced in rich medium, aceA disruption had no effect in vitro or on multiplication in macrophages or mouse spleens. The results suggest that B. abortus grows intracellularly using a limited supply of 6 C (and 5 C) sugars that is compensated by glutamate and possibly other amino acids entering the Krebs cycle without a critical role of the glyoxylate shunt.",
author = "Amaia Z{\'u}{\~n}iga-Ripa and Thibault Barbier and Raquel Conde-{\'A}lvarez and Estrella Mart{\'i}nez-G{\'o}mez and Leyre Palacios-Chaves and Yolanda Gil-Ram{\'i}rez and Grill{\'o}, {Mar{\'i}a Jes{\'u}s} and Jean-Jacques Letesson and Maite Iriarte and Ignacio Moriy{\'o}n",
note = "Copyright {\circledC} 2014, American Society for Microbiology. All Rights Reserved.",
year = "2014",
doi = "10.1128/JB.01663-14",
language = "English",
journal = "J Bacteriol",
issn = "0021-9193",
publisher = "American Society for Microbiology",

}

Brucella abortus depends on pyruvate phosphate dikinase and malic enzyme but not on Fbp and GlpX fructose-1,6-bisphosphatases for full virulence in laboratory models. / Zúñiga-Ripa, Amaia; Barbier, Thibault; Conde-Álvarez, Raquel; Martínez-Gómez, Estrella; Palacios-Chaves, Leyre; Gil-Ramírez, Yolanda; Grilló, María Jesús; Letesson, Jean-Jacques; Iriarte, Maite; Moriyón, Ignacio.

Dans: Journal of Bacteriology, 2014.

Résultats de recherche: Contribution à un journal/une revueArticle

TY - JOUR

T1 - Brucella abortus depends on pyruvate phosphate dikinase and malic enzyme but not on Fbp and GlpX fructose-1,6-bisphosphatases for full virulence in laboratory models

AU - Zúñiga-Ripa, Amaia

AU - Barbier, Thibault

AU - Conde-Álvarez, Raquel

AU - Martínez-Gómez, Estrella

AU - Palacios-Chaves, Leyre

AU - Gil-Ramírez, Yolanda

AU - Grilló, María Jesús

AU - Letesson, Jean-Jacques

AU - Iriarte, Maite

AU - Moriyón, Ignacio

N1 - Copyright © 2014, American Society for Microbiology. All Rights Reserved.

PY - 2014

Y1 - 2014

N2 - The brucellae are the etiological agents of brucellosis, a worldwide-distributed zoonosis. These bacteria are facultative intracellular parasites, and thus are able to adjust their metabolism to the extra and intracellular environments encountered during an infectious cycle. However, this aspect of Brucella biology is imperfectly known and the nutrients available in the intracellular niche are unknown. Here, we investigated the central pathways of C metabolism used by Brucella abortus by deleting the putative fructose-1,6-bisphosphatases (fbp and glpX), phosphoenolpyruvate carboxykinase (pckA), pyruvate phosphate dikinase (ppdK) and malic enzyme (mae) genes. In gluconeogenic but not in rich media, growth of mutants ΔppdK and Δmae was severely impaired and growth of the double Δfbp-ΔglpX mutant was reduced. In macrophages, only ΔppdK and Δmae showed reduced multiplication, and studies with ΔppdK confirmed that it reached the replicative niche. Similarly, only ΔppdK and Δmae were attenuated in mice, the former being cleared by week 10 and the latter persisting longer than 12 weeks. We also investigated the glyoxylate cycle. Although aceA (isocitrate lyase) promoter activity was enhanced in rich medium, aceA disruption had no effect in vitro or on multiplication in macrophages or mouse spleens. The results suggest that B. abortus grows intracellularly using a limited supply of 6 C (and 5 C) sugars that is compensated by glutamate and possibly other amino acids entering the Krebs cycle without a critical role of the glyoxylate shunt.

AB - The brucellae are the etiological agents of brucellosis, a worldwide-distributed zoonosis. These bacteria are facultative intracellular parasites, and thus are able to adjust their metabolism to the extra and intracellular environments encountered during an infectious cycle. However, this aspect of Brucella biology is imperfectly known and the nutrients available in the intracellular niche are unknown. Here, we investigated the central pathways of C metabolism used by Brucella abortus by deleting the putative fructose-1,6-bisphosphatases (fbp and glpX), phosphoenolpyruvate carboxykinase (pckA), pyruvate phosphate dikinase (ppdK) and malic enzyme (mae) genes. In gluconeogenic but not in rich media, growth of mutants ΔppdK and Δmae was severely impaired and growth of the double Δfbp-ΔglpX mutant was reduced. In macrophages, only ΔppdK and Δmae showed reduced multiplication, and studies with ΔppdK confirmed that it reached the replicative niche. Similarly, only ΔppdK and Δmae were attenuated in mice, the former being cleared by week 10 and the latter persisting longer than 12 weeks. We also investigated the glyoxylate cycle. Although aceA (isocitrate lyase) promoter activity was enhanced in rich medium, aceA disruption had no effect in vitro or on multiplication in macrophages or mouse spleens. The results suggest that B. abortus grows intracellularly using a limited supply of 6 C (and 5 C) sugars that is compensated by glutamate and possibly other amino acids entering the Krebs cycle without a critical role of the glyoxylate shunt.

U2 - 10.1128/JB.01663-14

DO - 10.1128/JB.01663-14

M3 - Article

C2 - 24936050

JO - J Bacteriol

JF - J Bacteriol

SN - 0021-9193

ER -