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The Trypanosome UDP-Glucose Pyrophosphorylase Is Imported by Piggybacking into Glycosomes, Where Unconventional Sugar Nucleotide Synthesis Takes Place

dc.rights.licenseopenen_US
hal.structure.identifierMicrobiologie Fondamentale et Pathogénicité [MFP]
dc.contributor.authorVILLAFRAZ, Oriana
hal.structure.identifierMicrobiologie Fondamentale et Pathogénicité [MFP]
hal.structure.identifierCentre de résonance magnétique des systèmes biologiques [CRMSB]
dc.contributor.authorBAUDOUIN, Hélène
hal.structure.identifierMicrobiologie Fondamentale et Pathogénicité [MFP]
hal.structure.identifierCentre de résonance magnétique des systèmes biologiques [CRMSB]
dc.contributor.authorMAZET, Muriel
hal.structure.identifierMetaboHUB-MetaToul
dc.contributor.authorKULYK, Hanna
hal.structure.identifierPlateforme Protéome [Bordeaux]
dc.contributor.authorDUPUY, Jean-William
hal.structure.identifierMicrobiologie Fondamentale et Pathogénicité [MFP]
dc.contributor.authorPINEDA, Erika
hal.structure.identifierInstitute for Advanced Biosciences / Institut pour l'Avancée des Biosciences (Grenoble) [IAB]
dc.contributor.authorBOTTÉ, Cyrille
dc.contributor.authorINAOKA, Daniel Ken
hal.structure.identifierMetaboHUB-MetaToul
hal.structure.identifierGeroscience and rejuvenation research center [RESTORE]
dc.contributor.authorPORTAIS, Jean-Charles
hal.structure.identifierMicrobiologie Fondamentale et Pathogénicité [MFP]
hal.structure.identifierCentre de résonance magnétique des systèmes biologiques [CRMSB]
dc.contributor.authorBRINGAUD, Frédéric
dc.date.accessioned2023-05-22T13:32:41Z
dc.date.available2023-05-22T13:32:41Z
dc.date.issued2021-06-29
dc.identifier.issn2161-2129en_US
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/182229
dc.description.abstractEnGlycosomes are peroxisome-related organelles of trypanosomatid para-sites containing metabolic pathways, such as glycolysis and biosynthesis of sugar nu-cleotides, usually present in the cytosol of other eukaryotes. UDP-glucose pyrophos-phorylase (UGP), the enzyme responsible for the synthesis of the sugar nucleotide UDP-glucose, is localized in the cytosol and glycosomes of the bloodstream and pro-cyclic trypanosomes, despite the absence of any known peroxisome-targeting signal (PTS1 and PTS2). The questions that we address here are (i) is the unusual glycoso-mal biosynthetic pathway of sugar nucleotides functional and (ii) how is the PTS-free UGP imported into glycosomes? We showed that UGP is imported into glycosomes by piggybacking on the glycosomal PTS1-containing phosphoenolpyruvate carboxy-kinase (PEPCK) and identified the domains involved in the UGP/PEPCK interaction. Proximity ligation assays revealed that this interaction occurs in 3 to 10% of glyco-somes, suggesting that these correspond to organelles competent for protein import. We also showed that UGP is essential for the growth of trypanosomes and that both the glycosomal and cytosolic metabolic pathways involving UGP are func-tional, since the lethality of the knockdown UGP mutant cell line ((RNAi)UGP, where RNAi indicates RNA interference) was rescued by expressing a recoded UGP (rUGP) in the organelle ((RNAi)UGP/(EXP)rUGP-GPDH, where GPDH is glycerol-3-phosphate dehy-drogenase). Our conclusion was supported by targeted metabolomic analyses (ion chromatography-high-resolution mass spectrometry [IC-HRMS]) showing that UDP-glucose is no longer detectable in the (RNAi)UGP mutant, while it is still produced in cells expressing UGP exclusively in the cytosol (PEPCK null mutant) or glycosomes ((RNAi)UGP/(EXP)rUGP-GPDH). Trypanosomatids are the only known organisms to have selected functional peroxisomal (glycosomal) sugar nucleotide biosynthetic pathways in addition to the canonical cytosolic ones.IMPORTANCE Unusual compartmentalization of metabolic pathways within organelles is one of the most enigmatic features of trypanosomatids. These unicellular eukar-yotes are the only organisms that sequestered glycolysis inside peroxisomes (glyco-somes), although the selective advantage of this compartmentalization is still not clear. Trypanosomatids are also unique for the glycosomal localization of enzymes of the sugar nucleotide biosynthetic pathways, which are also present in the cytosol. Here, we showed that the cytosolic and glycosomal pathways are functional. As in all other eukaryotes, the cytosolic pathways feed glycosylation reactions; however, the role of the duplicated glycosomal pathways is currently unknown. We also showed that one of these enzymes (UGP) is imported into glycosomes by piggybacking on another glycosomal enzyme (PEPCK); they are not functionally related. The UGP/PEPCK association is unique since all piggybacking examples reported to date involve functionally related interacting partners, which broadens the possible combinations of carrier-cargo proteins being imported as hetero-oligomers.
dc.description.sponsorshipVoies métaboliques glycosomales non glycolytiques: nouvelles fonctions pour le développement et la virulence des trypanosomes - ANR-15-CE15-0025en_US
dc.description.sponsorshipAlliance française contre les maladies parasitairesen_US
dc.language.isoENen_US
dc.rights.urihttp://creativecommons.org/licenses/by/
dc.subject.enperoxisomes
dc.subject.enpiggybacking
dc.subject.enprocyclic form
dc.subject.enTrypanosoma brucei
dc.subject.enUDP-glucose pyrophosphorylase
dc.subject.englycosomes
dc.title.enThe Trypanosome UDP-Glucose Pyrophosphorylase Is Imported by Piggybacking into Glycosomes, Where Unconventional Sugar Nucleotide Synthesis Takes Place
dc.typeArticle de revueen_US
dc.identifier.doi10.1128/mBio.00375-21en_US
dc.subject.halSciences du Vivant [q-bio]/Microbiologie et Parasitologieen_US
bordeaux.journalmBioen_US
bordeaux.volume12en_US
bordeaux.hal.laboratoriesMFP (Laboratoire Microbiologie Fondamentale et Pathogénicité) - UMR 5234en_US
bordeaux.issue3en_US
bordeaux.institutionCNRSen_US
bordeaux.institutionUniversité de Bordeaux
bordeaux.peerReviewedouien_US
bordeaux.inpressnonen_US
bordeaux.import.sourcehal
hal.identifierhal-03353350
hal.version1
hal.exportfalse
workflow.import.sourcehal
dc.rights.ccCC BYen_US
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=mBio&rft.date=2021-06-29&rft.volume=12&rft.issue=3&rft.eissn=2161-2129&rft.issn=2161-2129&rft.au=VILLAFRAZ,%20Oriana&BAUDOUIN,%20H%C3%A9l%C3%A8ne&MAZET,%20Muriel&KULYK,%20Hanna&DUPUY,%20Jean-William&rft.genre=article


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