Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines

Detalhes bibliográficos
Autor(a) principal: Moreira, Douglas de Souza
Data de Publicação: 2016
Outros Autores: Ferreira, Rafael Fernandes, Murta, Silvane Maria Fonseca
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/16303
Resumo: 2021-01-01
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spelling Moreira, Douglas de SouzaFerreira, Rafael FernandesMurta, Silvane Maria Fonseca2016-10-19T13:32:48Z2016-10-19T13:32:48Z2016MOREIRA, Douglas de Souza; FERREIRA, Rafael Fernandes; MURTA, Silvane Maria Fonseca. Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines. Exp Parasitol., n. 160, p. 60-66, 20160014-4894https://www.arca.fiocruz.br/handle/icict/1630310.1016/j.exppara.2015.12.009engElsevier LtdaLeishmanioseresistencia a drogasantioxidant defenseDrug resistanceLeishmaniaPteridine reductaseSbIIIMolecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania linesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article2021-01-01Fundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, BrazilPteridine reductase (PTR1) is an NADPH-dependent reductase that participates in the salvage of pteridines, which are essential to maintain growth of Leishmania. In this study, we performed the molecular characterization of ptr1 gene in wild-type (WTS) and SbIII-resistant (SbR) lines from Leishmania guyanensis (Lg), Leishmania amazonensis (La), Leishmania braziliensis (Lb) and Leishmania infantum (Li), evaluating the chromosomal location, mRNA levels of the ptr1 gene and PTR1 protein expression. PFGE results showed that the ptr1 gene is located in a 797 kb chromosomal band in all Leishmania lines analyzed. Interestingly, an additional chromosomal band of 1070 kb was observed only in LbSbR line. Northern blot results showed that the levels of ptr1 mRNA are increased in the LgSbR, LaSbR and LbSbR lines. Western blot assays using the polyclonal anti-LmPTR1 antibody demonstrated that PTR1 protein is more expressed in the LgSbR, LaSbR and LbSbR lines compared to their respective WTS counterparts. Nevertheless, no difference in the level of mRNA and protein was observed between the LiWTS and LiSbR lines. Functional analysis of PTR1 enzyme was performed to determine whether the overexpression of ptr1 gene in the WTS L. braziliensis and L. infantum lines would change the SbIII-resistance phenotype of transfected parasites. Western blot results showed that the expression level of PTR1 protein was increased in the transfected parasites compared to the non-transfected ones. IC50 analysis revealed that the overexpression of ptr1 gene in the WTS L. braziliensis line increased 2-fold the SbIII-resistance phenotype compared to the non-transfected counterpart. Furthermore, the overexpression of ptr1 gene in the WTS L. infantum line did not change the SbIII-resistance phenotype. These results suggest that the PTR1 enzyme may be implicated in the SbIII-resistance phenotype in L. braziliensis line.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/16303/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALve_Moreira_Douglas_Molecular characterization_CPqRR_2016.pdfve_Moreira_Douglas_Molecular characterization_CPqRR_2016.pdfapplication/pdf797543https://www.arca.fiocruz.br/bitstream/icict/16303/2/ve_Moreira_Douglas_Molecular%20characterization_CPqRR_2016.pdf28d1ddca4c4f15ca23cbe35d4962ae8bMD52TEXTve_Moreira_Douglas_Molecular characterization_CPqRR_2016.pdf.txtve_Moreira_Douglas_Molecular characterization_CPqRR_2016.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
title Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
spellingShingle Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
Moreira, Douglas de Souza
Leishmaniose
resistencia a drogas
antioxidant defense
Drug resistance
Leishmania
Pteridine reductase
SbIII
title_short Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
title_full Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
title_fullStr Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
title_full_unstemmed Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
title_sort Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
author Moreira, Douglas de Souza
author_facet Moreira, Douglas de Souza
Ferreira, Rafael Fernandes
Murta, Silvane Maria Fonseca
author_role author
author2 Ferreira, Rafael Fernandes
Murta, Silvane Maria Fonseca
author2_role author
author
dc.contributor.author.fl_str_mv Moreira, Douglas de Souza
Ferreira, Rafael Fernandes
Murta, Silvane Maria Fonseca
dc.subject.other.pt_BR.fl_str_mv Leishmaniose
resistencia a drogas
topic Leishmaniose
resistencia a drogas
antioxidant defense
Drug resistance
Leishmania
Pteridine reductase
SbIII
dc.subject.en.pt_BR.fl_str_mv antioxidant defense
Drug resistance
Leishmania
Pteridine reductase
SbIII
description 2021-01-01
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-10-19T13:32:48Z
dc.date.available.fl_str_mv 2016-10-19T13:32:48Z
dc.date.issued.fl_str_mv 2016
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv MOREIRA, Douglas de Souza; FERREIRA, Rafael Fernandes; MURTA, Silvane Maria Fonseca. Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines. Exp Parasitol., n. 160, p. 60-66, 2016
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/16303
dc.identifier.issn.pt_BR.fl_str_mv 0014-4894
dc.identifier.eissn.none.fl_str_mv 10.1016/j.exppara.2015.12.009
identifier_str_mv MOREIRA, Douglas de Souza; FERREIRA, Rafael Fernandes; MURTA, Silvane Maria Fonseca. Molecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines. Exp Parasitol., n. 160, p. 60-66, 2016
0014-4894
10.1016/j.exppara.2015.12.009
url https://www.arca.fiocruz.br/handle/icict/16303
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier Ltda
publisher.none.fl_str_mv Elsevier Ltda
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/16303/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/16303/2/ve_Moreira_Douglas_Molecular%20characterization_CPqRR_2016.pdf
https://www.arca.fiocruz.br/bitstream/icict/16303/3/ve_Moreira_Douglas_Molecular%20characterization_CPqRR_2016.pdf.txt
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