Marjorie Montenegro1,2, Claudia Cuervo1, Constanza Cardenas3, Silvia Duarte1, Jenny R Díaz1, M Carmen Thomas2, Manuel C Lopez2, Concepcion J Puerta1,+
1Pontificia Universidad Javeriana, Facultad de Ciencias, Departamento de Microbiología, Laboratorio de Parasitología Molecular, Bogotá, Colombia
2Consejo Superior de Investigaciones Científicas, Instituto de Parasitología y Biomedicina López Neyra, Granada, Spain
3Pontificia Universidad Católica de Valparaíso, Núcleo de Biotecnología Curauma, Valparaíso, Chile
Trypanosomatid type I nitroreductases (NTRs), i.e., mitochondrial enzymes that metabolise nitroaromatic pro-drugs, are essential for parasite growth, infection, and survival. Here, a type I NTR of non-virulent protozoan Trypanosoma rangeli is described and compared to those of other trypanosomatids. The NTR gene was isolated from KP1(+) and KP1(-) strains, and its corresponding transcript and 5’ untranslated region (5’UTR) were determined. Bioinformatics analyses and nitro-drug activation assays were also performed. The results indicated that the type I NTR gene is present in both KP1(-) and KP1(+) strains, with 98% identity. However, the predicted subcellular localisation of the protein differed among the strains (predicted as mitochondrial in the KP1(+) strain). Comparisons of the domains and 3D structures of the NTRs with those of orthologs demonstrated that the nitroreductase domain of T. rangeli NTR is conserved across all the strains, including the residues involved in the interaction with the FMN cofactor and in the tertiary structure characteristics of this oxidoreductase protein family. mRNA processing and expression were also observed. In addition, T. rangeli was shown to be sensitive to benznidazole and nifurtimox in a concentration-dependent manner. In summary, T. rangeli appears to have a newly discovered functional type I NTR.
Financial support: Colciencias Research (Project No. RC-595-2009), Pontificia Universidad Javeriana Research (Project No. 6728). MM and JRD were supported by the programs Jóvenes Investigadores e Innovadores 2011 and 2016 from Colciencias; MCL and MCT were supported by grants SAF2016-81003R and SAF2016-80998, respectively, from Plan Nacional I+D+i (MICINN) and RD12/0018/0021 from ISCIII-RETIC (MICINN), Spain, and FEDER.
Received 12 December 2016
Accepted 22 March 2017