Metabolic selection of a homologous recombination-mediated gene loss protects Trypanosoma brucei from ROS production by glycosomal fumarate reductase
| Autor: | Wargnies, Marion, Plazolles, Nicolas, Schenk, Robin, Villafraz, Oriana, Dupuy, Jean-William, Biran, Marc, Bachmaier, Sabine, Baudouin, Hélène, Clayton, Christine, Boshart, Michael, Bringaud, Frédéric |
|---|---|
| Přispěvatelé: | Université de Bordeaux (UB), Centre de résonance magnétique des systèmes biologiques (CRMSB), Centre National de la Recherche Scientifique (CNRS)-Université de Bordeaux (UB), Microbiologie cellulaire et moléculaire et pathogénicité (MCMP), Université Bordeaux Segalen - Bordeaux 2-Centre National de la Recherche Scientifique (CNRS), Ludwig-Maximilians-Universität München (LMU), Plateforme Protéome [Bordeaux], Centre Génomique Fonctionnelle Bordeaux [Bordeaux] (CGFB), Institut Polytechnique de Bordeaux-Université de Bordeaux Ségalen [Bordeaux 2]-Institut Polytechnique de Bordeaux-Université de Bordeaux Ségalen [Bordeaux 2], Universität Heidelberg [Heidelberg] |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
phosphoenolpyruvate carboxykinase (PEPCK)
Trypanosoma brucei brucei homologous recombination Microbodies ROS reactive oxygen species PPDK pyruvate phosphate dikinase positive selection Flavins [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry Molecular Biology/Genomics [q-bio.GN] [SDV.MP.PAR]Life Sciences [q-bio]/Microbiology and Parasitology/Parasitology Cells Cultured ComputingMilieux_MISCELLANEOUS trypanosoma Cytb5R domain reactive oxygen species parasite differentiation PEP phosphoenolpyruvate PCF procyclic form ACN acetonitrile covalent flavinylation FRD fumarate reductase Succinate Dehydrogenase Glucose genomic rearrangement PEPCK phosphoenolpyruvate carboxykinase NADH-dependent fumarate reductase (FRD) Research Article |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2021, 296, pp.100548. ⟨10.1016/j.jbc.2021.100548⟩ The Journal of Biological Chemistry |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1016/j.jbc.2021.100548⟩ |
| Popis: | International audience; The genome of trypanosomatids rearranges by using repeated sequences as platforms for amplification or deletion of genomic segments. These stochastic recombination events have a direct impact on gene dosage and foster the selection of adaptive traits in response to environmental pressure. We provide here such an example by showing that the phosphoenolpyruvate carboxykinase (PEPCK) gene knockout (Δpepck) leads to the selection of a deletion event between two tandemly arranged fumarate reductase (FRDg and FRDm2) genes to produce a chimeric FRDg-m2 gene in the Δpepck∗ cell line. FRDg is expressed in peroxisome-related organelles, named glycosomes, expression of FRDm2 has not been detected to date, and FRDg-m2 is nonfunctional and cytosolic. Re-expression of FRDg significantly impaired growth of the Δpepck∗ cells, but FRD enzyme activity was not required for this negative effect. Instead, glycosomal localization as well as the covalent flavinylation motif of FRD is required to confer growth retardation and intracellular accumulation of reactive oxygen species (ROS). The data suggest that FRDg, similar to Escherichia coli FRD, can generate ROS in a flavin-dependent process by transfer of electrons from NADH to molecular oxygen instead of fumarate when the latter is unavailable, as in the Δpepck background. Hence, growth retardation is interpreted as a consequence of increased production of ROS, and rearrangement of the FRD locus liberates Δpepck∗ cells from this obstacle. Interestingly, intracellular production of ROS has been shown to be required to complete the parasitic cycle in the insect vector, suggesting that FRDg may play a role in this process |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|