Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/16303
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dc.contributor.authorMoreira, Douglas de Souza
dc.contributor.authorFerreira, Rafael Fernandes
dc.contributor.authorMurta, Silvane Maria Fonseca
dc.date.accessioned2016-10-19T13:32:48Z
dc.date.available2016-10-19T13:32:48Z
dc.date.issued2016
dc.identifier.citationMOREIRA, 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.issn0014-4894
dc.identifier.urihttps://www.arca.fiocruz.br/handle/icict/16303
dc.language.isoeng
dc.publisherElsevier Ltda
dc.rightsrestricted access
dc.subject.otherLeishmaniose
dc.subject.otherresistencia a drogas
dc.titleMolecular characterization and functional analysis of pteridine reductase in wild-type and antimony-resistant Leishmania lines
dc.typeArticle
dc.description.abstractenPteridine 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.
dc.creator.affilliationFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brazil
dc.creator.affilliationFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brazil
dc.creator.affilliationFundação Oswaldo Cruz. Centro de Pesquisas Rene Rachou. Laboratorio de Parasitologia Celular e Molecular. Belo Horizonte, MG, Brazil
dc.subject.enantioxidant defense
dc.subject.enDrug resistance
dc.subject.enLeishmania
dc.subject.enPteridine reductase
dc.subject.enSbIII
dc.identifier.eissn10.1016/j.exppara.2015.12.009
Appears in Collections:MG - IRR - Artigos de Periódicos

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