Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery

Autores
Krakovka, Sascha; Ranjbarian, Farahnaz; Luján, Lucas Agustín; Saura, Alicia; Larsen, Nicolai B.; Jiménez González, Alejandro; Reggenti, Anna; Lujan, Hugo Daniel; Svärd, Staffan G.; Hofer, Anders
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Giardiasis is a diarrheal disease caused by the unicellular parasite Giardia intestinalis, for which metronidazole is the main treatment option. The parasite is dependent on exogenous deoxyribonucleosides for DNA replication and thus is also potentially vulnerable to deoxyribonucleoside analogs. Here, we characterized the G. intestinalis thymidine kinase, a divergent member of the thymidine kinase 1 family that consists of two weakly homologous parts within one polypeptide. We found that the recombinantly expressed enzyme is monomeric, with 100-fold higher catalytic efficiency for thymidine compared to its second-best substrate, deoxyuridine, and is furthermore subject to feedback inhibition by dTTP. This efficient substrate discrimination is in line with the lack of thymidylate synthase and dUTPase in the parasite, which makes deoxy-UMP a dead-end product that is potentially harmful if converted to deoxy-UTP. We also found that the antiretroviral drug azidothymidine (AZT) was an equally good substrate as thymidine and was active against WT as well as metronidazole-resistant G. intestinalis trophozoites. This drug inhibited DNA synthesis in the parasite and efficiently decreased cyst production in vitro, which suggests that it could reduce infectivity. AZT also showed a good effect in G. intestinalis?infected gerbils, reducing both the number of trophozoites in the small intestine and the number of viable cysts in the stool. Taken together, these results suggest that the absolute dependency of the parasite on thymidine kinase for its DNA synthesis can be exploited by AZT, which has promise as a future medication effective against metronidazole-refractory giardiasis.
Fil: Krakovka, Sascha. Uppsala Universitet; Suecia
Fil: Ranjbarian, Farahnaz. Universidad de Umea; Suecia
Fil: Luján, Lucas Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Saura, Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Larsen, Nicolai B.. No especifíca;
Fil: Jiménez González, Alejandro. Uppsala Universitet; Suecia
Fil: Reggenti, Anna. Universidad de Umea; Suecia
Fil: Lujan, Hugo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Svärd, Staffan G.. Uppsala Universitet; Suecia
Fil: Hofer, Anders. Universidad de Umea; Suecia
Materia
AZIDOTHYMIDINE
DEOXYNUCLEOSIDE KINASE
DEOXYNUCLEOSIDE SALVAGE
DEOXYRIBONUCLEOSIDE KINASE
DEOXYRIBONUCLEOSIDE SALVAGE
GIARDIA DUODENUM
GIARDIA INTESTINALIS
GIARDIA LAMBLIA
THYMIDINE KINASE
ZIDOVUDINE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/222423
Acceder
id CONICETDig_9f4387914305283fff19ded120f45bfb
oai_identifier_str oai:ri.conicet.gov.ar:11336/222423
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discoveryKrakovka, SaschaRanjbarian, FarahnazLuján, Lucas AgustínSaura, AliciaLarsen, Nicolai B.Jiménez González, AlejandroReggenti, AnnaLujan, Hugo DanielSvärd, Staffan G.Hofer, AndersAZIDOTHYMIDINEDEOXYNUCLEOSIDE KINASEDEOXYNUCLEOSIDE SALVAGEDEOXYRIBONUCLEOSIDE KINASEDEOXYRIBONUCLEOSIDE SALVAGEGIARDIA DUODENUMGIARDIA INTESTINALISGIARDIA LAMBLIATHYMIDINE KINASEZIDOVUDINEhttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Giardiasis is a diarrheal disease caused by the unicellular parasite Giardia intestinalis, for which metronidazole is the main treatment option. The parasite is dependent on exogenous deoxyribonucleosides for DNA replication and thus is also potentially vulnerable to deoxyribonucleoside analogs. Here, we characterized the G. intestinalis thymidine kinase, a divergent member of the thymidine kinase 1 family that consists of two weakly homologous parts within one polypeptide. We found that the recombinantly expressed enzyme is monomeric, with 100-fold higher catalytic efficiency for thymidine compared to its second-best substrate, deoxyuridine, and is furthermore subject to feedback inhibition by dTTP. This efficient substrate discrimination is in line with the lack of thymidylate synthase and dUTPase in the parasite, which makes deoxy-UMP a dead-end product that is potentially harmful if converted to deoxy-UTP. We also found that the antiretroviral drug azidothymidine (AZT) was an equally good substrate as thymidine and was active against WT as well as metronidazole-resistant G. intestinalis trophozoites. This drug inhibited DNA synthesis in the parasite and efficiently decreased cyst production in vitro, which suggests that it could reduce infectivity. AZT also showed a good effect in G. intestinalis?infected gerbils, reducing both the number of trophozoites in the small intestine and the number of viable cysts in the stool. Taken together, these results suggest that the absolute dependency of the parasite on thymidine kinase for its DNA synthesis can be exploited by AZT, which has promise as a future medication effective against metronidazole-refractory giardiasis.Fil: Krakovka, Sascha. Uppsala Universitet; SueciaFil: Ranjbarian, Farahnaz. Universidad de Umea; SueciaFil: Luján, Lucas Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; ArgentinaFil: Saura, Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; ArgentinaFil: Larsen, Nicolai B.. No especifíca;Fil: Jiménez González, Alejandro. Uppsala Universitet; SueciaFil: Reggenti, Anna. Universidad de Umea; SueciaFil: Lujan, Hugo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; ArgentinaFil: Svärd, Staffan G.. Uppsala Universitet; SueciaFil: Hofer, Anders. Universidad de Umea; SueciaAmerican Society for Biochemistry and Molecular Biology2022-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/222423Krakovka, Sascha; Ranjbarian, Farahnaz; Luján, Lucas Agustín; Saura, Alicia; Larsen, Nicolai B.; et al.; Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 6; 6-2022; 1-160021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2022.102028info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:23:22Zoai:ri.conicet.gov.ar:11336/222423instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:23:23.217CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
title Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
spellingShingle Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
Krakovka, Sascha
AZIDOTHYMIDINE
DEOXYNUCLEOSIDE KINASE
DEOXYNUCLEOSIDE SALVAGE
DEOXYRIBONUCLEOSIDE KINASE
DEOXYRIBONUCLEOSIDE SALVAGE
GIARDIA DUODENUM
GIARDIA INTESTINALIS
GIARDIA LAMBLIA
THYMIDINE KINASE
ZIDOVUDINE
title_short Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
title_full Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
title_fullStr Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
title_full_unstemmed Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
title_sort Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery
dc.creator.none.fl_str_mv Krakovka, Sascha
Ranjbarian, Farahnaz
Luján, Lucas Agustín
Saura, Alicia
Larsen, Nicolai B.
Jiménez González, Alejandro
Reggenti, Anna
Lujan, Hugo Daniel
Svärd, Staffan G.
Hofer, Anders
author Krakovka, Sascha
author_facet Krakovka, Sascha
Ranjbarian, Farahnaz
Luján, Lucas Agustín
Saura, Alicia
Larsen, Nicolai B.
Jiménez González, Alejandro
Reggenti, Anna
Lujan, Hugo Daniel
Svärd, Staffan G.
Hofer, Anders
author_role author
author2 Ranjbarian, Farahnaz
Luján, Lucas Agustín
Saura, Alicia
Larsen, Nicolai B.
Jiménez González, Alejandro
Reggenti, Anna
Lujan, Hugo Daniel
Svärd, Staffan G.
Hofer, Anders
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv AZIDOTHYMIDINE
DEOXYNUCLEOSIDE KINASE
DEOXYNUCLEOSIDE SALVAGE
DEOXYRIBONUCLEOSIDE KINASE
DEOXYRIBONUCLEOSIDE SALVAGE
GIARDIA DUODENUM
GIARDIA INTESTINALIS
GIARDIA LAMBLIA
THYMIDINE KINASE
ZIDOVUDINE
topic AZIDOTHYMIDINE
DEOXYNUCLEOSIDE KINASE
DEOXYNUCLEOSIDE SALVAGE
DEOXYRIBONUCLEOSIDE KINASE
DEOXYRIBONUCLEOSIDE SALVAGE
GIARDIA DUODENUM
GIARDIA INTESTINALIS
GIARDIA LAMBLIA
THYMIDINE KINASE
ZIDOVUDINE
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Giardiasis is a diarrheal disease caused by the unicellular parasite Giardia intestinalis, for which metronidazole is the main treatment option. The parasite is dependent on exogenous deoxyribonucleosides for DNA replication and thus is also potentially vulnerable to deoxyribonucleoside analogs. Here, we characterized the G. intestinalis thymidine kinase, a divergent member of the thymidine kinase 1 family that consists of two weakly homologous parts within one polypeptide. We found that the recombinantly expressed enzyme is monomeric, with 100-fold higher catalytic efficiency for thymidine compared to its second-best substrate, deoxyuridine, and is furthermore subject to feedback inhibition by dTTP. This efficient substrate discrimination is in line with the lack of thymidylate synthase and dUTPase in the parasite, which makes deoxy-UMP a dead-end product that is potentially harmful if converted to deoxy-UTP. We also found that the antiretroviral drug azidothymidine (AZT) was an equally good substrate as thymidine and was active against WT as well as metronidazole-resistant G. intestinalis trophozoites. This drug inhibited DNA synthesis in the parasite and efficiently decreased cyst production in vitro, which suggests that it could reduce infectivity. AZT also showed a good effect in G. intestinalis?infected gerbils, reducing both the number of trophozoites in the small intestine and the number of viable cysts in the stool. Taken together, these results suggest that the absolute dependency of the parasite on thymidine kinase for its DNA synthesis can be exploited by AZT, which has promise as a future medication effective against metronidazole-refractory giardiasis.
Fil: Krakovka, Sascha. Uppsala Universitet; Suecia
Fil: Ranjbarian, Farahnaz. Universidad de Umea; Suecia
Fil: Luján, Lucas Agustín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Saura, Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Larsen, Nicolai B.. No especifíca;
Fil: Jiménez González, Alejandro. Uppsala Universitet; Suecia
Fil: Reggenti, Anna. Universidad de Umea; Suecia
Fil: Lujan, Hugo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas. Universidad Católica de Córdoba. Centro de Investigación y Desarrollo en Inmunología y Enfermedades Infecciosas; Argentina
Fil: Svärd, Staffan G.. Uppsala Universitet; Suecia
Fil: Hofer, Anders. Universidad de Umea; Suecia
description Giardiasis is a diarrheal disease caused by the unicellular parasite Giardia intestinalis, for which metronidazole is the main treatment option. The parasite is dependent on exogenous deoxyribonucleosides for DNA replication and thus is also potentially vulnerable to deoxyribonucleoside analogs. Here, we characterized the G. intestinalis thymidine kinase, a divergent member of the thymidine kinase 1 family that consists of two weakly homologous parts within one polypeptide. We found that the recombinantly expressed enzyme is monomeric, with 100-fold higher catalytic efficiency for thymidine compared to its second-best substrate, deoxyuridine, and is furthermore subject to feedback inhibition by dTTP. This efficient substrate discrimination is in line with the lack of thymidylate synthase and dUTPase in the parasite, which makes deoxy-UMP a dead-end product that is potentially harmful if converted to deoxy-UTP. We also found that the antiretroviral drug azidothymidine (AZT) was an equally good substrate as thymidine and was active against WT as well as metronidazole-resistant G. intestinalis trophozoites. This drug inhibited DNA synthesis in the parasite and efficiently decreased cyst production in vitro, which suggests that it could reduce infectivity. AZT also showed a good effect in G. intestinalis?infected gerbils, reducing both the number of trophozoites in the small intestine and the number of viable cysts in the stool. Taken together, these results suggest that the absolute dependency of the parasite on thymidine kinase for its DNA synthesis can be exploited by AZT, which has promise as a future medication effective against metronidazole-refractory giardiasis.
publishDate 2022
dc.date.none.fl_str_mv 2022-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/222423
Krakovka, Sascha; Ranjbarian, Farahnaz; Luján, Lucas Agustín; Saura, Alicia; Larsen, Nicolai B.; et al.; Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 6; 6-2022; 1-16
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/222423
identifier_str_mv Krakovka, Sascha; Ranjbarian, Farahnaz; Luján, Lucas Agustín; Saura, Alicia; Larsen, Nicolai B.; et al.; Giardia intestinalis thymidine kinase is a high-affinity enzyme crucial for DNA synthesis and an exploitable target for drug discovery; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 6; 6-2022; 1-16
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2022.102028
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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