De novo biosynthesis of sterols and fatty acids in the Trypanosoma brucei procyclic form: Carbon source preferences and metabolic flux redistributions
| Autor: | Millerioux, Yoann, Mazet, Muriel, Bouyssou, Guillaume, Allmann, Stefan, Kiema, Tiila-Riikka, Bertiaux, Eloise, Fouillen, Laetitia, Thapa, Chandan, Biran, Marc, Plazolles, Nicolas, Dittrich-Domergue, Franziska, Crouzols, Aline, Wierenga, Rik, Rotureau, Brice, MOREAU, Patrick, Bringaud, Frédéric |
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| Přispěvatelé: | Microbiologie Fondamentale et Pathogénicité (MFP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Centre de résonance magnétique des systèmes biologiques (CRMSB), Université de Bordeaux (UB)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de biogenèse membranaire (LBM), University of Oulu, Biologie cellulaire des Trypanosomes - Trypanosome Cell Biology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), This research 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 (grant number ANR-2010-BLAN-1319-02) of the ANR-BLANC-2010 call and GLYCONOV (grant number ANR-15-CE15-0025-01) of the 'Générique' 2015 call, the Laboratoire d’Excellence (LabEx) ParaFrap (grant number ANR-11-LABX-0024), the Institut Pasteur and by the European Commission FP7 Marie Curie Initial Training Network 'ParaMet' (grant number 290080). EB was funded by a doctoral fellowship from French National Ministry for Research and Technology (doctoral school CDV515)., 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-11-LABX-0024,ParaFrap,Alliance française contre les maladies parasitaires(2011), European Project: 290080,EC:FP7:PEOPLE,FP7-PEOPLE-2011-ITN,PARAMET(2012), Résonance magnétique des systèmes biologiques (RMSB), Laboratoire Microorganismes : Génome et Environnement (LMGE), Université Blaise Pascal - Clermont-Ferrand 2 (UBP)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Centre National de la Recherche Scientifique (CNRS), Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Biologie cellulaire des Trypanosomes, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB), Rotureau, Brice, 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, 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 |
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Threonine
[SDV]Life Sciences [q-bio] blood-sream forms Acetates Biochemistry molecular characterization Gene Knockout Techniques dependent enzyme Glucose Metabolism proline metabolism MESH: Animals Amino Acids MESH: Threonine lcsh:QH301-705.5 MESH: Gene Knockout Techniques Protozoans MESH: Tsetse Flies Organic Compounds Fatty Acids Monosaccharides Eukaryota cell-cycle acetyl-coa energy-metabolism lipbiosyntesis leishmania-mexicana succinate coa-transferase MESH: Mevalonic Acid Lipids MESH: Gene Expression Regulation [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry Molecular Biology/Biomolecules [q-bio.BM] MESH: Fatty Acids [SDV] Life Sciences [q-bio] MESH: Glucose Sterols Chemistry [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology Physical Sciences Carbohydrate Metabolism MESH: Acetates [SDV.MP.PAR] Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Research Article lcsh:Immunologic diseases. Allergy Trypanosoma Proline Tsetse Flies Trypanosoma brucei brucei Carbohydrates Mevalonic Acid MESH: Carbon MESH: Insect Vectors Biosynthesis [SDV.MP.PRO]Life Sciences [q-bio]/Microbiology and Parasitology/Protistology MESH: Alcohol Oxidoreductases Acetyl Coenzyme A Acetyltransferases Leucine Hydroxyl Amino Acids MESH: Acyl Coenzyme A Animals [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology [SDV.MP] Life Sciences [q-bio]/Microbiology and Parasitology MESH: Proline Organic Chemistry Organisms Chemical Compounds Biology and Life Sciences Proteins MESH: Trypanosoma brucei brucei MESH: Acetyltransferases Carbon Parasitic Protozoans Insect Vectors Alcohol Oxidoreductases Glucose Metabolism MESH: Leucine Gene Expression Regulation Aliphatic Amino Acids lcsh:Biology (General) MESH: Sterols Acyl Coenzyme A lcsh:RC581-607 MESH: Acetyl Coenzyme A |
| Zdroj: | PLoS Pathogens PLoS Pathogens, 2018, 14 (5), pp.e1007116. ⟨10.1371/journal.ppat.1007116⟩ PLoS Pathogens, Vol 14, Iss 5, p e1007116 (2018) Plos Pathogens 5 (14), . (2018) PLoS Pathogens, Public Library of Science, 2018, 14 (5), pp.e1007116. ⟨10.1371/journal.ppat.1007116⟩ |
| ISSN: | 1553-7366 1553-7374 |
| DOI: | 10.1371/journal.ppat.1007116⟩ |
| Popis: | De novo biosynthesis of lipids is essential for Trypanosoma brucei, a protist responsible for the sleeping sickness. Here, we demonstrate that the ketogenic carbon sources, threonine, acetate and glucose, are precursors for both fatty acid and sterol synthesis, while leucine only contributes to sterol production in the tsetse fly midgut stage of the parasite. Degradation of these carbon sources into lipids was investigated using a combination of reverse genetics and analysis of radio-labelled precursors incorporation into lipids. For instance, (i) deletion of the gene encoding isovaleryl-CoA dehydrogenase, involved in the leucine degradation pathway, abolished leucine incorporation into sterols, and (ii) RNAi-mediated down-regulation of the SCP2-thiolase gene expression abolished incorporation of the three ketogenic carbon sources into sterols. The SCP2-thiolase is part of a unidirectional two-step bridge between the fatty acid precursor, acetyl-CoA, and the precursor of the mevalonate pathway leading to sterol biosynthesis, 3-hydroxy-3-methylglutaryl-CoA. Metabolic flux through this bridge is increased either in the isovaleryl-CoA dehydrogenase null mutant or when the degradation of the ketogenic carbon sources is affected. We also observed a preference for fatty acids synthesis from ketogenic carbon sources, since blocking acetyl-CoA production from both glucose and threonine abolished acetate incorporation into sterols, while incorporation of acetate into fatty acids was increased. Interestingly, the growth of the isovaleryl-CoA dehydrogenase null mutant, but not that of the parental cells, is interrupted in the absence of ketogenic carbon sources, including lipids, which demonstrates the essential role of the mevalonate pathway. We concluded that procyclic trypanosomes have a strong preference for fatty acid versus sterol biosynthesis from ketogenic carbon sources, and as a consequence, that leucine is likely to be the main source, if not the only one, used by trypanosomes in the infected insect vector digestive tract to feed the mevalonate pathway. Author summary In this study, we have (i) determined the carbon sources used by the Trypanosoma brucei procyclic insect form to feed the essential lipid biosynthetic pathways, (ii) further characterized the metabolic pathways leading to their degradation into acetyl-CoA (fatty acid precursor) and 3-hydroxy-3-methylglutaryl-CoA (sterol precursor) and (iii) showed that reduction of the ketogenic carbon sources degradation, favors their incorporation into fatty acids, instead of sterols. This fatty acid preference is compensated by an increase of leucine incorporation into sterols, which highlights the parasite adaptation capacity regarding carbon source availability by modulating the metabolic flux between branches within the network. This metabolic flexibility is particularly relevant for the insect stages of trypanosomes that evolve in the midgut and the salivary glands of their blood-feeding insect vector. One may also consider that, the metabolic flow redistribution towards the mevalonate pathway (sterol production) described in vitro also occurs in vivo, depending on the carbon source composition of the tsetse fly micro-environment, which may considerably vary along the digestive tract and depending on the fly feeding status, as well as in the other infected fly organs. |
| Databáze: | OpenAIRE |
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