Contribution of Pyruvate Phosphate Dikinase in the Maintenance of the Glycosomal ATP/ADP Balance in the Trypanosoma brucei Procyclic Form

Deramchia, Kamel; Morand, Pauline; Biran, Marc; Millerioux, Yoann; Mazet, Muriel; Wargnies, Marion; Franconi, Jean‐Michel; Bringaud, Frédéric

doi:10.1074/jbc.m114.567230

Cited by 38 publications

(56 citation statements)

References 56 publications

(95 reference statements)

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“…Characterization of the FRDg gene closed a long-standing debate about the existence of bona fide soluble NADH-dependent FRD in trypanosomes Turrens, 1999) and strengthened Gest's hypothesis, as glucose metabolism in the trypanosome glycosomes closely resembles his succinic fermentation model. Indeed, inclusion of the glycosomal succinate branch in the procyclic trypanosomes releases PEP molecules used to produce ATP in the cytosol through pyruvate kinase activity, which has been described as essential for the viability of the procyclic form (Morris et al, 2002;Coustou et al, 2003;Deramchia et al, 2014). The trypanosome FRD enzymes were the first soluble NADH-dependent FRD characterized, although the concept might be more widely applicable as soluble NADH-dependent FRD activity was reported in the parasite Trichomonas foetus (Müller, 2003) and the prokaryote Hydrogenobacter thermophilus (Miura et al, 2008).…”

Section: Succinic Fermentationmentioning

confidence: 99%

“…pyruvate phosphate dikinase (PPDK, step 5) working in the gluconeogenic direction (Bringaud et al, 1998), PEPCK (step 1) and malate dehydrogenase (step 2), may operate without any impact on the glycosomal ATP/ADP balance as ATP molecules consumed by PPDK are regenerated by PEPCK. Its utilization is certainly exceptional, as the procyclic cells growing in standard glucose-rich conditions preferentially use PPDK in the glycolytic direction to participate in the maintenance of this glycosomal ATP/ADP balance (Deramchia et al, 2014). The relevance of this temporarily used hypothetical cycle is its ability to produce cytosolic NADPH sustained by NADH production in the glycosomes, which may compensate for the absence of cytosolic (and mitochondrial) transhydrogenases in trypanosomatids.…”

Section: Connections Between the Acetate And Succinate Branches Provimentioning

confidence: 99%

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Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways

Bringaud

Biran

Millerioux

et al. 2015

Molecular Microbiology

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SummaryNumerous eukaryotes have developed specific metabolic traits that are not present in extensively studied model organisms. For instance, the procyclic insect form of Trypanosoma brucei, a parasite responsible for sleeping sickness in its mammalianspecific bloodstream form, metabolizes glucose into excreted succinate and acetate through pathways with unique features. Succinate is primarily produced from glucose-derived phosphoenolpyruvate in peroxisome-like organelles, also known as glycosomes, by a soluble NADH-dependent fumarate reductase only described in trypanosomes so far. Acetate is produced in the mitochondrion of the parasite from acetyl-CoA by a CoA-transferase, which forms an ATP-producing cycle with succinylCoA synthetase. The role of this cycle in ATP production was recently demonstrated in procyclic trypanosomes and has only been proposed so far for anaerobic organisms, in addition to trypanosomatids. We review how nuclear magnetic resonance spectrometry can be used to analyze the metabolic network perturbed by deletion (knockout) or downregulation (RNAi) of the candidate genes involved in these two particular metabolic pathways of procyclic trypanosomes. The role of succinate and acetate production in trypanosomes is discussed, as well as the connections between the succinate and acetate branches, which increase the metabolic flexibility probably required by the parasite to deal with environmental changes such as oxidative stress.

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Section: Succinic Fermentationmentioning

confidence: 99%

Section: Connections Between the Acetate And Succinate Branches Provimentioning

confidence: 99%

Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways

Bringaud

Biran

Millerioux

et al. 2015

Molecular Microbiology

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“…In conclusion, we describe here a new mechanism for the regulation of nutrient utilization based on the competition between two enzymes (kinases) for a common substrate (ATP). The sequestration of the two kinases in the glycosomes is key to this mechanism since these peroxisome-like organelles show limited or no nucleotide exchange with the cytosol at a metabolic timescale (Deramchia et al, 2014; Haanstra et al, 2014). Hence, the ATP pool available to glycosomal kinases is limited, offering a situation where significant excess of one kinase (here GK) can abolish almost completely the flux through the other one (HK).…”

Section: Resultsmentioning

confidence: 99%

“Metabolic contest”, a new way to control carbon source preference

Allmann¹,

Wargnies²,

Cahoreau³

et al. 2019

Preprint

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SUMMARYMicroorganisms must make the right choice for nutrient consumption to adapt to their changing environment. As a consequence, bacteria and yeasts have developed regulatory mechanisms involving nutrient sensing and signaling, allowing to redirect cell metabolism to maximize the consumption of an energy-efficient carbon source. Here, we report a new mechanism, named “metabolic contest”, for regulating the use of carbon sources without nutrient sensing and signaling. In contrast to most microorganisms, trypanosomes show a glycerol-to-glucose preference that depends on the combination of three conditions: (i) the sequestration of both metabolic pathways in the same subcellular compartment, here in the peroxisomal-like organelles named glycosomes; (ii) the competition for the same substrate, here ATP, with the first enzymatic step of the glycerol and glucose metabolic pathways being both ATP-dependent (glycerol kinase and hexokinase, respectively) and (iii) an unbalanced activity between the competing enzymes, here the glycerol kinase activity being ~80-fold higher than the hexokinase activity.

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“…To investigate the biological role of TbSLP in the procyclic form of T. brucei , the TbSLP gene was deleted by gene replacement as previously described . Replacement of both TbSLP alleles by the puromycin and blasticidin selectable markers in the procyclic EATRO1125.T7T background was confirmed by PCR (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Replacement of the TbSLP gene by the blasticidin and puromycin resistance markers via homologous recombination was performed with DNA fragments containing the resistance marker gene flanked by TbSLP UTR sequences, as described previously . Briefly, the pGEMt plasmid was used to clone a HpaI DNA fragment containing either the blasticidin or puromycin resistance marker gene preceded by the TbSLP 5'‐UTR fragment (758 bp) and followed by the TbSLP 3'‐UTR fragment (754 bp).…”

Section: Methodsmentioning

confidence: 99%

The SCP2-thiolase-like protein (SLP) ofTrypanosoma bruceiis an enzyme involved in lipid metabolism

et al. 2016

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Bioinformatics studies have shown that the genomes of trypanosomatid species each encode one SCP2-thiolase-like protein (SLP), which is characterized by having the YDCF thiolase sequence fingerprint of the Cβ2-Cα2 loop. SLPs are only encoded by the genomes of these parasitic protists and not by those of mammals, including human. Deletion of the Trypanosoma brucei SLP gene (TbSLP) increases the doubling time of procyclic T. brucei and causes a 5-fold reduction of de novo sterol biosynthesis from glucose- and acetate-derived acetyl-CoA. Fluorescence analyses of EGFP-tagged TbSLP expressed in the parasite located the TbSLP in the mitochondrion. The crystal structure of TbSLP (refined at 1.75 Å resolution) confirms that TbSLP has the canonical dimeric thiolase fold. In addition, the structures of the TbSLP-acetoacetyl-CoA (1.90 Å) and TbSLP-malonyl-CoA (2.30 Å) complexes reveal that the two oxyanion holes of the thiolase active site are preserved. TbSLP binds malonyl-CoA tightly (Kd 90 µM), acetoacetyl-CoA moderately (Kd 0.9 mM) and acetyl-CoA and CoA very weakly. TbSLP possesses low malonyl-CoA decarboxylase activity. Altogether, the data show that TbSLP is a mitochondrial enzyme involved in lipid metabolism. Proteins 2016; 84:1075-1096. © 2016 Wiley Periodicals, Inc.

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Contribution of Pyruvate Phosphate Dikinase in the Maintenance of the Glycosomal ATP/ADP Balance in the Trypanosoma brucei Procyclic Form

Cited by 38 publications

References 56 publications

Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways

Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways

“Metabolic contest”, a new way to control carbon source preference

The SCP2-thiolase-like protein (SLP) ofTrypanosoma bruceiis an enzyme involved in lipid metabolism

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