Glycerol supports growth of the Trypanosoma brucei bloodstream forms in the absence of glucose: Analysis of metabolic adaptations on glycerol-rich conditions
| Autor: | Cyril Masante, Jean-William Dupuy, Stefan Allmann, Loïc Rivière, Frédéric Bringaud, Brice Rotureau, Arnaud Mourier, Marc Biran, Erika Pineda, Magali Thonnus, Hanna Kulyk, Muriel Mazet, Jean-Charles Portais, Edern Cahoreau |
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| Přispěvatelé: | Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Résonance magnétique des systèmes biologiques (RMSB), Institut de biochimie et génétique cellulaires (IBGC), Laboratoire d'Ingénierie des Systèmes Biologiques et des Procédés (LISBP), Centre National de la Recherche Scientifique (CNRS)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Recherche Agronomique (INRA), Centre Génomique Fonctionnelle Bordeaux [Bordeaux] (CGFB), Institut Polytechnique de Bordeaux-Université de Bordeaux Ségalen [Bordeaux 2], Biologie cellulaire des Trypanosomes, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de résonance magnétique des systèmes biologiques (CRMSB), Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB), Biologie cellulaire des Trypanosomes - Trypanosome Cell Biology, We thank Paul A. Michels (Edinburgh, Scotland) for the anti-FBPase and anti-GK immune sera, and Peter Overath (Tubingen, Germany) and Minu Chaudhuri (Nashville, TN, USA) for the anti-ISG75 and anti-PAO antibodies, respectively., ANR-10-BLAN-1319,ACETOTRYP,Metabolisme de l'acetyl-CoA et de l'acetate chez les trypanosomes: identification de nouvelles voies métaboliques spécifiques aux parasites(2010), ANR-15-CE15-0025,GLYCONOV,Voies métaboliques glycosomales non glycolytiques: nouvelles fonctions pour le développement et la virulence des trypanosomes(2015), ANR-11-LABX-0024,ParaFrap,Alliance française contre les maladies parasitaires(2011), Microbiologie Fondamentale et Pathogénicité (MFP), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Bordeaux, CNRS, European Project: 290080,EC:FP7:PEOPLE,FP7-PEOPLE-2011-ITN,PARAMET(2012), Bidault, Floran, BLANC - Metabolisme de l'acetyl-CoA et de l'acetate chez les trypanosomes: identification de nouvelles voies métaboliques spécifiques aux parasites - - ACETOTRYP2010 - ANR-10-BLAN-1319 - BLANC - VALID, Voies métaboliques glycosomales non glycolytiques: nouvelles fonctions pour le développement et la virulence des trypanosomes - - GLYCONOV2015 - ANR-15-CE15-0025 - AAPG2015 - VALID, Laboratoires d'excellence - Alliance française contre les maladies parasitaires - - ParaFrap2011 - ANR-11-LABX-0024 - LABX - VALID, A systematic analysis of parasite metabolism - from metabolism to intervention - PARAMET - - EC:FP7:PEOPLE2012-06-01 - 2016-05-31 - 290080 - VALID, Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Université de Bordeaux, the Agence Nationale de la Recherche (ANR) through grants ACETOTRYP of the ANR-BLANC-2010 call and GLYCONOV of the 'Générique' call, the Laboratoire d’Excellence (LabEx) ParaFrap ANR-11-LABX-0024 and the ParaMet PhD programme of Marie Curie Initial Training Network, ANR-11-LABX-0024/11-LABX-0024,ParaFrap,Alliance française contre les maladies parasitaires(2011) |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Glycerol Glycerol kinase [SDV]Life Sciences [q-bio] Succinic Acid Biochemistry chemistry.chemical_compound Tandem Mass Spectrometry Enzyme Chemistry lcsh:QH301-705.5 Protozoans chemistry.chemical_classification Glucose metabolism Organic Compounds Monomers Monosaccharides Microbiology and Parasitology Eukaryota Microbiologie et Parasitologie Mitochondria [SDV] Life Sciences [q-bio] Chemistry [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Adipose Tissue Physical Sciences Carbohydrate Metabolism Oxygen metabolism Glycolysis [SDV.MP.PAR] Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Research Article lcsh:Immunologic diseases. Allergy Cell Physiology Alternative oxidase Trypanosoma Trypanosoma brucei brucei Immunology Carbohydrates Carbohydrate metabolism Microbiology [SDV.MP.PRO]Life Sciences [q-bio]/Microbiology and Parasitology/Protistology Cell Line Phosphates 03 medical and health sciences Enzyme metabolism Virology Genetics Metabolomics Hexose [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology Molecular Biology Dihydroxyacetone phosphate Cell metabolism Organic Chemistry Gluconeogenesis Organisms Chemical Compounds Biology and Life Sciences Cell Biology Metabolism Polymer Chemistry Parasitic Protozoans Culture Media 030104 developmental biology Glucose lcsh:Biology (General) chemistry Enzymology Parasitology maladie du sommeil lcsh:RC581-607 trypanosoma brucei |
| Zdroj: | PLoS Pathogens PLoS Pathogens, Public Library of Science, 2018, 14 (11), pp.e1007412. ⟨10.1371/journal.ppat.1007412⟩ Plos Pathogens 11 (14), 25 p.. (2018) PLoS Pathogens, 2018, 14 (11), pp.e1007412. ⟨10.1371/journal.ppat.1007412⟩ PLoS Pathogens, Vol 14, Iss 11, p e1007412 (2018) |
| ISSN: | 1553-7366 1553-7374 |
| Popis: | The bloodstream forms of Trypanosoma brucei (BSF), the parasite protist causing sleeping sickness, primarily proliferate in the blood of their mammalian hosts. The skin and adipose tissues were recently identified as additional major sites for parasite development. Glucose was the only carbon source known to be used by bloodstream trypanosomes to feed their central carbon metabolism, however, the metabolic behaviour of extravascular tissue-adapted parasites has not been addressed yet. Since the production of glycerol is an important primary function of adipocytes, we have adapted BSF trypanosomes to a glucose-depleted but glycerol-rich culture medium (CMM_Glyc/GlcNAc) and compared their metabolism and proteome to those of parasites grown in standard glucose-rich conditions (CMM_Glc). BSF were shown to consume 2-folds more oxygen per consumed carbon unit in CMM_Glyc/GlcNAc and were 11.5-times more sensitive to SHAM, a specific inhibitor of the plant-like alternative oxidase (TAO), which is the only mitochondrial terminal oxidase expressed in BSF. This is consistent with (i) the absolute requirement of the mitochondrial respiratory activity to convert glycerol into dihydroxyacetone phosphate, as deduced from the updated metabolic scheme and (ii) with the 1.8-fold increase of the TAO expression level compared to the presence of glucose. Proton NMR analysis of excreted end products from glycerol and glucose metabolism showed that these two carbon sources are metabolised through the same pathways, although the contributions of the acetate and succinate branches are more important in the presence of glycerol than glucose (10.2% versus 3.4% of the excreted end products, respectively). In addition, metabolomic analyses by mass spectrometry showed that, in the absence of glucose, 13C-labelled glycerol was incorporated into hexose phosphates through gluconeogenesis. As expected, RNAi-mediated down-regulation of glycerol kinase expression abolished glycerol metabolism and was lethal for BSF grown in CMM_Glyc/GlcNAc. Interestingly, BSF have adapted their metabolism to grow in CMM_Glyc/GlcNAc by concomitantly increasing their rate of glycerol consumption and decreasing that of glucose. However, the glycerol kinase activity was 7.8-fold lower in CMM_Glyc/GlcNAc, as confirmed by both western blotting and proteomic analyses. This suggests that the huge excess in glycerol kinase that is not absolutely required for glycerol metabolism, might be used for another yet undetermined non-essential function in glucose rich-conditions. Altogether, these data demonstrate that BSF trypanosomes are well-adapted to glycerol-rich conditions that could be encountered by the parasite in extravascular niches, such as the skin and adipose tissues. Author summary Until very recently, the bloodstream forms (BSF) of the Trypanosoma brucei group species have been considered to propagate exclusively in the mammalian fluids, including the blood, the lymphatic network and the cerebrospinal fluid. All these fluids are rich in glucose, which is widely considered by the scientific community as the only carbon source used by the parasite to feed its central carbon metabolism and its ATP production. Here, we show for the first time that the BSF trypanosomes efficiently grow in glucose-free conditions as long as glycerol is supplied. The raison d'être of this capacity developed by BSF trypanosomes to grow in glycerol-rich conditions regardless of the glucose concentration, including in glucose-free conditions, is not yet understood. However, the recent discovery that trypanosomes colonize and proliferate in the skin and the adipose tissues of their mammalian hosts may provide a rational explanation for the development of a glycerol-based metabolism in BSF. Indeed, the adipocytes composing adipose tissues and also abundantly present in subcutaneous layers excrete large amounts of glycerol produced from the catabolism of glucose and triglycerides. We also show that BSF trypanosomes adapted to glucose-depleted conditions activate gluconeogenesis to produce the essential hexose phosphates from glycerol metabolism. Interestingly, the constitutive expression of the key gluconeogenic enzyme fructose-1,6-bisphosphatase, which is not used for glycolysis, suggests that BSF trypanosomes maintained in the standard glucose-rich medium are pre-adapted to glucose-depleted conditions. This further strengthens the new paradigm that BSF trypanosomes can use glycerol in tissues producing this carbon source, such as the skin the adipose tissues. |
| Databáze: | OpenAIRE |
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