Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse

Gao, Jianpeng; Tian, Mingxing; Bao, Yanqing; Liu, Jiameng; Ding, Chan; Wang, Shaohui; Li, Tao; Yu, Shuang

doi:10.1186/s13567-016-0372-7

Cited by 20 publications

(18 citation statements)

References 25 publications

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“…The importance of B. abortus glycolysis in infection is supported by data showing that the deletion of pyruvate kinase, which presumably renders the strain unable to execute the last step of glycolysis, results in attenuation in macrophage and mouse infection models (40). However, this mutation is pleiotropic and may result in attenuation for a variety of reasons.…”

Section: Discussionmentioning

confidence: 58%

Brucella abortus Induces a Warburg Shift in Host Metabolism That Is Linked to Enhanced Intracellular Survival of the Pathogen

2017

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Intracellular bacterial pathogens exploit host cell resources to replicate and survive inside the host. Targeting these host systems is one promising approach to developing novel antimicrobials to treat intracellular infections. We show that human macrophage-like cells infected with Brucella abortus undergo a metabolic shift characterized by attenuated tricarboxylic acid cycle metabolism, reduced amino acid consumption, altered mitochondrial localization, and increased lactate production. This shift to an aerobic glycolytic state resembles the Warburg effect, a change in energy production that is well described in cancer cells and also occurs in activated inflammatory cells. B. abortus efficiently uses lactic acid as its sole carbon and energy source and requires the ability to metabolize lactate for normal survival in human macrophage-like cells. We demonstrate that chemical inhibitors of host glycolysis and lactate production do not affect in vitro growth of B. abortus in axenic culture but decrease its survival in the intracellular niche. Our data support a model in which infection shifts host metabolism to a Warburg-like state, and B. abortus uses this change in metabolism to promote intracellular survival. Pharmacological perturbation of these features of host cell metabolism may be a useful strategy to inhibit infection by intracellular pathogens.IMPORTANCE Brucella spp. are intracellular bacterial pathogens that cause disease in a range of mammals, including livestock. Transmission from livestock to humans is common and can lead to chronic human disease. Human macrophage-like cells infected with Brucella abortus undergo a Warburg-like metabolic shift to an aerobic glycolytic state where the host cells produce lactic acid and have reduced amino acid catabolism. We provide evidence that the pathogen can exploit this change in host metabolism to support growth and survival in the intracellular niche. Drugs that inhibit this shift in host cell metabolism inhibit intracellular replication and decrease the survival of B. abortus in an in vitro infection model; these drugs may be broadly useful therapeutics for intracellular infections.KEYWORDS infection, Warburg effect, lactate, therapeutics, antimetabolite drug, 3-bromopyruvic acid, NHI-2, 2-deoxy-D-glucose, brucellosis, antibiotic, metabolism, zoonotic infection M acrophages can be activated by pathogen-associated proinflammatory molecules, such as lipopolysaccharide (LPS). Classically activated (i.e., M1) macrophages undergo a major metabolic shift in which the tricarboxylic acid (TCA) cycle is downregulated, and energy production transitions from oxidative phosphorylation to the less efficient process of aerobic glycolysis (1, 2). This change in metabolism, known as the Warburg effect (3), is a well-described metabolic feature of cancer cells (4).

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Section: Discussionmentioning

confidence: 58%

Brucella abortus Induces a Warburg Shift in Host Metabolism That Is Linked to Enhanced Intracellular Survival of the Pathogen

2017

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“…PykM and PpdK make independent contributions to the virulence of B. abortus 2308 in mice. The data presented here and elsewhere (23,27) strongly suggest that the pyruvate kinase PykM and pyruvate phosphate dikinase PpdK make different contributions to glucose metabolism in Brucella, with PykM being required for wild-type catabolism of this sugar and PpdK being needed for its synthesis from gluconeogenic substrates. Both of these enzymes are required for the wild-type virulence of B. abortus 2308 in BALB/c mice (23,27).…”

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confidence: 52%

The Manganese-Dependent Pyruvate Kinase PykM Is Required for Wild-Type Glucose Utilization by Brucella abortus 2308 and Its Virulence in C57BL/6 Mice

et al. 2018

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Pyruvate kinase plays a central role in glucose catabolism in bacteria, and efficient utilization of this hexose has been linked to the virulence of Brucella strains in mice. The brucellae produce a single pyruvate kinase which is an ortholog of the Bradyrhizobium manganese (Mn)-dependent pyruvate kinase PykM. A biochemical analysis of the Brucella pyruvate kinase and phenotypic analysis of a Brucella abortus mutant defective in high-affinity Mn import indicate that this enzyme is an authentic PykM ortholog which functions as a Mn-dependent enzyme in vivo. The loss of PykM has a negative impact on the capacity of the parental 2308 strain to utilize glucose, fructose, and galactose but not on its ability to utilize ribose, xylose, arabinose, or erythritol, and a pykM mutant displays significant attenuation in C57BL/6 mice. Although the enzyme pyruvate phosphate dikinase (PpdK) can substitute for the loss of pyruvate kinase in some bacteria and is also an important virulence determinant in Brucella, a phenotypic analysis of B. abortus 2308 and isogenic pykM, ppdK, and pykM ppdK mutants indicates that PykM and PpdK make distinctly different contributions to carbon metabolism and virulence in these bacteria. IMPORTANCE Mn plays a critical role in the physiology and virulence of Brucella strains, and the results presented here suggest that one of the important roles that the high-affinity Mn importer MntH plays in the pathogenesis of these strains is supporting the function of the Mn-dependent kinase PykM. A better understanding of how the brucellae adapt their physiology and metabolism to sustain their intracellular persistence in host macrophages will provide knowledge that can be used to design improved strategies for preventing and treating brucellosis, a disease that has a significant impact on both the veterinary and public health communities worldwide. Citation Pitzer JE, Zeczycki TN, Baumgartner JE, Martin DW, Roop RM, II. 2018. The manganesedependent pyruvate kinase PykM is required for wild-type glucose utilization by Brucella abortus 2308 and its virulence in C57BL/6 mice. J Bacteriol 200:e00471-18. https://doi.org/10 .1128/JB. Downloaded fromin vitro cultivation (5), but for others, this metal only appears to be required when the bacteria are subjected to an environmental stress, such as an exposure to reactive oxygen species (ROS) (6). Mn-dependent superoxide dismutases such as SodA, for instance, are important cytoplasmic antioxidants in bacteria (7), and in some cases, bacteria can substitute Mn for Fe in cellular proteins that do not require the redox activity of the latter metal for their function as a means of protecting these proteins from damage mediated by ROS via Fenton chemistry (6).Brucella strains produce a single high-affinity Mn transporter, MntH, and a phenotypic analysis of an isogenic mntH mutant derived from B. abortus 2308 indicates that Mn plays an important role in the basic physiology of these bacteria (8). The B. abortus mntH mutant, for instance, displays delayed growth compared to...

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“…The relevance of a Brucella -induced increase in aerobic glycolysis and lactate production to in vivo infection biology in natural hosts remains unclear. The importance of B. abortus glycolysis in infection is supported by data showing that deletion of pyruvate kinase, which presumably renders the strain unable to execute the last step of glycolysis, results in attenuation in macrophage and mouse infection models (40). However, this mutation is pleiotropic and may result in attenuation for a variety of reasons.…”

Section: Discussionmentioning

confidence: 98%

Brucella abortusinduces a Warburg shift in host metabolism that is linked to enhanced intracellular survival of the pathogen

Czyż

Willett

Crosson

2017

Preprint

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Brucella abortus induces a Warburg shift in host metabolism that is linked toHoward Taylor Ricketts Laboratory, University of Chicago, Argonne National that is well-described in cancer cells, and also occurs in activated inflammatory cells. B. 5abortus efficiently uses lactic acid as its sole carbon and energy source and requires the 2 6 ability to metabolize lactate for normal survival in human macrophage-like cells. We 25, 2017; 3 host cells produce lactic acid and have reduced amino acid catabolism. We provide defenses by increasing concentrations of ROS and reactive nitrogen intermediates (6). response to support infection. 4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http://dx.doi.org/10.1101/120527 doi: bioRxiv preprint first posted online Mar.4.0 International license peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was not . http:/ RESULTS 2Brucella abortus infection of human macrophage-like cells disrupts mitochondrial To characterize the effect of intracellular colonization by B. abortus on human were cultivated in minimal essential medium in the presence of central metabolic substrates specifically selected to measure mitochondrial function.

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Pyruvate kinase is necessary for Brucella abortus full virulence in BALB/c mouse

Cited by 20 publications

References 25 publications

Brucella abortus Induces a Warburg Shift in Host Metabolism That Is Linked to Enhanced Intracellular Survival of the Pathogen

Brucella abortus Induces a Warburg Shift in Host Metabolism That Is Linked to Enhanced Intracellular Survival of the Pathogen

The Manganese-Dependent Pyruvate Kinase PykM Is Required for Wild-Type Glucose Utilization by Brucella abortus 2308 and Its Virulence in C57BL/6 Mice

Brucella abortusinduces a Warburg shift in host metabolism that is linked to enhanced intracellular survival of the pathogen

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