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Pyrimidine salvage enzymes are essential for de novo biosynthesis of Deoxypyrimidine nucleotides in Trypanosoma brucei

2016Artigo científico Acesso aberto
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dc.contributor.authorLeija, Christopher
dc.contributor.authorRijo-Ferreira, Filipa
dc.contributor.authorKinch, Lisa N.
dc.contributor.authorGrishin, Nick V.
dc.contributor.authorNischan, Nicole
dc.contributor.authorKohler, Jennifer J.
dc.contributor.authorHu, Zeping
dc.contributor.authorPhillips, Margaret A.
dc.date.accessioned2022-01-18T15:16:00Z
dc.date.available2022-01-18T15:16:00Z
dc.date.issued2016
dc.description© 2016 Leija et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.pt_PT
dc.description.abstractThe human pathogenic parasite Trypanosoma brucei possess both de novo and salvage routes for the biosynthesis of pyrimidine nucleotides. Consequently, they do not require salvageable pyrimidines for growth. Thymidine kinase (TK) catalyzes the formation of dTMP and dUMP and is one of several salvage enzymes that appear redundant to the de novo pathway. Surprisingly, we show through analysis of TK conditional null and RNAi cells that TK is essential for growth and for infectivity in a mouse model, and that a catalytically active enzyme is required for its function. Unlike humans, T. brucei and all other kinetoplastids lack dCMP deaminase (DCTD), which provides an alternative route to dUMP formation. Ectopic expression of human DCTD resulted in full rescue of the RNAi growth phenotype and allowed for selection of viable TK null cells. Metabolite profiling by LC-MS/MS revealed a buildup of deoxypyrimidine nucleosides in TK depleted cells. Knockout of cytidine deaminase (CDA), which converts deoxycytidine to deoxyuridine led to thymidine/deoxyuridine auxotrophy. These unexpected results suggested that T. brucei encodes an unidentified 5'-nucleotidase that converts deoxypyrimidine nucleotides to their corresponding nucleosides, leading to their dead-end buildup in TK depleted cells at the expense of dTTP pools. Bioinformatics analysis identified several potential candidate genes that could encode 5'-nucleotidase activity including an HD-domain protein that we show catalyzes dephosphorylation of deoxyribonucleotide 5'-monophosphates. We conclude that TK is essential for synthesis of thymine nucleotides regardless of whether the nucleoside precursors originate from the de novo pathway or through salvage. Reliance on TK in the absence of DCTD may be a shared vulnerability among trypanosomatids and may provide a unique opportunity to selectively target a diverse group of pathogenic single-celled eukaryotes with a single drug.pt_PT
dc.description.sponsorshipThis work was supported by National Institutes of Health (grants AI078962 and AI034432) to MAP (https://www.niaid.nih.gov) and (grant GM007062) to CL (https://www.nigms.nih. gov), the Welch Foundation (grant I-1257) to MAP and (grant I-1686) to JJK (http://www.welch1.org), and Fundac ̧ão para a Ciência e Tecnologia (FCT, Portugal) SFRH/BD/51286/2010 (http://www.fct.pt) to FRF.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationPLoS Pathog. 2016 Nov 7;12(11):e1006010pt_PT
dc.identifier.doi10.1371/journal.ppat.1006010pt_PT
dc.identifier.eissn1553-7374
dc.identifier.issn1553-7366
dc.identifier.urihttp://hdl.handle.net/10451/50879
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherPLOSpt_PT
dc.relation.publisherversionhttps://journals.plos.org/plospathogens/pt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.titlePyrimidine salvage enzymes are essential for de novo biosynthesis of Deoxypyrimidine nucleotides in Trypanosoma bruceipt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/SFRH/SFRH%2FBD%2F51286%2F2010/PT
oaire.citation.issue11pt_PT
oaire.citation.titlePLOS Pathogenspt_PT
oaire.citation.volume12pt_PT
oaire.fundingStreamSFRH
person.familyNameRijo-Ferreira
person.givenNameFilipa
person.identifier750870
person.identifier.ciencia-id7E1A-ED0E-636C
person.identifier.orcid0000-0002-4003-020X
person.identifier.scopus-author-id56261883100
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication56fd4f5e-09a9-4739-bf21-6165f21cf944
relation.isAuthorOfPublication.latestForDiscovery56fd4f5e-09a9-4739-bf21-6165f21cf944
relation.isProjectOfPublication99755d1c-477a-4303-85a2-f7265f48b340
relation.isProjectOfPublication.latestForDiscovery99755d1c-477a-4303-85a2-f7265f48b340

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