title: Combining reverse genetics and nuclear magnetic resonance-based metabolomics unravels trypanosome-specific metabolic pathways.
abstract: Numerous 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 mammalian-specific 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 succinyl-CoA 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.
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Bringaud
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Biran
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Millerioux
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Marion
authors.initials: M
authors.lastname: Wargnies
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Stefan
authors.initials: S
authors.lastname: Allmann
authors.affiliation: Centre de Résonance Magnétique des Systèmes Biologiques, UMR-5536 Université de Bordeaux, CNRS, 146 rue Léo Saignat, 33076, Bordeaux, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Mazet
mesh_terms: D000085:Acetates / Q000378:metabolism; D000105:Acetyl Coenzyme A / Q000235:genetics / Q000378:metabolism; D000214:Acyl Coenzyme A / Q000235:genetics / Q000378:metabolism; D000818:Animals; D055786:Gene Knockout Techniques; D005947:Glucose / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D053858:Metabolic Networks and Pathways / Q000235:genetics / Q000502:physiology; D055432:Metabolomics / Q000379:methods; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D018384:Oxidative Stress / Q000235:genetics / Q000502:physiology; D034622:RNA Interference; D059386:Reverse Genetics; D019802:Succinic Acid / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
chemical_list: D000085:Acetates; D000214:Acyl Coenzyme A; D000105:Acetyl Coenzyme A; D019802:Succinic Acid; C012046:succinyl-coenzyme A; D005947:Glucose