Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity

[Background] Malaria remains as a major global problem, being one of the infectious diseases that engender highest mortality across the world. Due to the appearance of resistance and the lack of an effective vaccine, the search of novel anti-malarials is required. Deoxyuridine 5′-triphosphate nucleo...

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Autores: Pérez-Moreno, Guiomar, Sánchez-Carrasco, Paula, Ruiz-Pérez, Luis Miguel, Johansson, N.G., Müller, Sylke, Baragaña, Beatriz, Hampton, Shahienaz, Gilbert, Ian H., Kaiser, Marcel, Sarkar, Sandipan, Pandurangan, Thiyagamurthy, Kumar, Vijeesh, González-Pacanowska, Dolores
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/196266
Acceso en línea:http://hdl.handle.net/10261/196266
Access Level:acceso abierto
Palabra clave:lasmodium falciparum
Deoxyuridine 5′-triphosphate nucleotido-hydrolase
5′-tritylated deoxyuridine analogues
Mode of action
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spelling Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activityPérez-Moreno, GuiomarSánchez-Carrasco, PaulaRuiz-Pérez, Luis MiguelJohansson, N.G.Müller, SylkeBaragaña, BeatrizHampton, ShahienazGilbert, Ian H.Kaiser, MarcelSarkar, SandipanPandurangan, ThiyagamurthyKumar, VijeeshGonzález-Pacanowska, Doloreslasmodium falciparumDeoxyuridine 5′-triphosphate nucleotido-hydrolase5′-tritylated deoxyuridine analoguesMode of action[Background] Malaria remains as a major global problem, being one of the infectious diseases that engender highest mortality across the world. Due to the appearance of resistance and the lack of an effective vaccine, the search of novel anti-malarials is required. Deoxyuridine 5′-triphosphate nucleotido-hydrolase (dUTPase) is responsible for the hydrolysis of dUTP to dUMP within the parasite and has been proposed as an essential step in pyrimidine metabolism by providing dUMP for thymidylate biosynthesis. In this work, efforts to validate dUTPase as a drug target in Plasmodium falciparum are reported.[Methods] To investigate the role of PfdUTPase in cell survival different strategies to generate knockout mutants were used. For validation of PfdUTPase as the intracellular target of four inhibitors of the enzyme, mutants overexpressing PfdUTPase and HsdUTPase were created and the IC50 for each cell line with each compound was determined. The effect of these compounds on dUTP and dTTP levels from P. falciparum was measured using a DNA polymerase assay. Detailed localization studies by indirect immunofluorescence microscopy and live cell imaging were also performed using a cell line overexpressing a Pfdut-GFP fusion protein.[Results] Different attempts of disruption of the dut gene of P. falciparum were unsuccessful while a 3′ replacement construct could recombine correctly in the locus suggesting that the enzyme is essential. The four 5′-tritylated deoxyuridine analogues described are potent inhibitors of the P. falciparum dUTPase and exhibit antiplasmodial activity. Overexpression of the Plasmodium and human enzymes conferred resistance against selective compounds, providing chemical validation of the target and confirming that indeed dUTPase inhibition is involved in anti-malarial activity. In addition, incubation with these inhibitors was associated with a depletion of the dTTP pool corroborating the central role of dUTPase in dTTP synthesis. PfdUTPase is mainly localized in the cytosol.[Conclusion] These results strongly confirm the pivotal and essential role of dUTPase in pyrimidine biosynthesis of P. falciparum intraerythrocytic stages.This work was funded by the Junta de Andalucía (BIO-199); the Plan Nacional de Investigación Científica, Instituto de Salud Carlos III-Subdirección General de Redes y Centros de Investigación Cooperativa-Red de Investigación Cooperativa en Enfermedades Tropicales (RICET: RD16/0027/0014); The European Union (contract 037587); and the Plan Nacional (SAF2016-79957-R).Peer reviewedBioMed CentralJunta de AndalucíaMinisterio de Economía y Competitividad (España)European CommissionConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2019201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/196266reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2016-79957-Rhttps://doi.org/10.1186/s12936-019-3025-2Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1962662026-05-22T06:33:51Z
dc.title.none.fl_str_mv Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
title Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
spellingShingle Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
Pérez-Moreno, Guiomar
lasmodium falciparum
Deoxyuridine 5′-triphosphate nucleotido-hydrolase
5′-tritylated deoxyuridine analogues
Mode of action
title_short Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
title_full Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
title_fullStr Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
title_full_unstemmed Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
title_sort Validation of Plasmodium falciparum dUTPase as the target of 5′-tritylated deoxyuridine analogues with anti-malarial activity
dc.creator.none.fl_str_mv Pérez-Moreno, Guiomar
Sánchez-Carrasco, Paula
Ruiz-Pérez, Luis Miguel
Johansson, N.G.
Müller, Sylke
Baragaña, Beatriz
Hampton, Shahienaz
Gilbert, Ian H.
Kaiser, Marcel
Sarkar, Sandipan
Pandurangan, Thiyagamurthy
Kumar, Vijeesh
González-Pacanowska, Dolores
author Pérez-Moreno, Guiomar
author_facet Pérez-Moreno, Guiomar
Sánchez-Carrasco, Paula
Ruiz-Pérez, Luis Miguel
Johansson, N.G.
Müller, Sylke
Baragaña, Beatriz
Hampton, Shahienaz
Gilbert, Ian H.
Kaiser, Marcel
Sarkar, Sandipan
Pandurangan, Thiyagamurthy
Kumar, Vijeesh
González-Pacanowska, Dolores
author_role author
author2 Sánchez-Carrasco, Paula
Ruiz-Pérez, Luis Miguel
Johansson, N.G.
Müller, Sylke
Baragaña, Beatriz
Hampton, Shahienaz
Gilbert, Ian H.
Kaiser, Marcel
Sarkar, Sandipan
Pandurangan, Thiyagamurthy
Kumar, Vijeesh
González-Pacanowska, Dolores
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Junta de Andalucía
Ministerio de Economía y Competitividad (España)
European Commission
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv lasmodium falciparum
Deoxyuridine 5′-triphosphate nucleotido-hydrolase
5′-tritylated deoxyuridine analogues
Mode of action
topic lasmodium falciparum
Deoxyuridine 5′-triphosphate nucleotido-hydrolase
5′-tritylated deoxyuridine analogues
Mode of action
description [Background] Malaria remains as a major global problem, being one of the infectious diseases that engender highest mortality across the world. Due to the appearance of resistance and the lack of an effective vaccine, the search of novel anti-malarials is required. Deoxyuridine 5′-triphosphate nucleotido-hydrolase (dUTPase) is responsible for the hydrolysis of dUTP to dUMP within the parasite and has been proposed as an essential step in pyrimidine metabolism by providing dUMP for thymidylate biosynthesis. In this work, efforts to validate dUTPase as a drug target in Plasmodium falciparum are reported.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/196266
url http://hdl.handle.net/10261/196266
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2016-79957-R
https://doi.org/10.1186/s12936-019-3025-2
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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