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Combining reverse genetics and nuclear magnetic resonance‐based metabolomics unravels trypanosome‐specific metabolic pathways
Author(s) -
Bringaud Frédéric,
Biran Marc,
Millerioux Yoann,
Wargnies Marion,
Allmann Stefan,
Mazet Muriel
Publication year - 2015
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/mmi.12990
Subject(s) - biology , fumarate reductase , metabolic pathway , trypanosoma brucei , biochemistry , citric acid cycle , mitochondrion , metabolomics , organelle , phosphoenolpyruvate carboxykinase , peroxisome , gene , microbiology and biotechnology , enzyme , succinate dehydrogenase , bioinformatics
Summary Numerous eukaryotes have developed specific metabolic traits that are not present in extensively studied model organisms. For instance, the procyclic insect form of T rypanosoma 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.