Overexpression of superoxide dismutase in Trypanosoma cruzi results in increased sensitivity to the trypanocidal agents gentian violet and benznidazole.
The parasitic protozoan Trypanosoma cruzi is exposed to toxic oxygen metabolites which arise from drug metabolism or immune mechanisms, in addition to those produced by endogenous processes. Identification and functional analysis of parasite enzymes which confer protection against oxidative stress is therefore of importance. To investigate the role of T. cruzi superoxide dismutase (SOD) we transfected epimastigotes with an expression vector containing a putative Fe-SOD gene homologue and achieved overexpression of enzyme activity (5-8 fold). Inhibition studies carried out on the partially purified enzyme revealed azide and H2O2 sensitivity and cyanide insensitivity, the profile expected of an Fe-isoform. Phenotypic analysis of transformed parasites showed that they were more susceptible than control cells to growth inhibition by the trypanocidal drug benznidazole and by gentian violet, an agent which can be used to decontaminate blood supplies in endemic areas. These results may reflect an imbalance in the antioxidant defences of the parasite produced as a result of overexpression of Fe-SOD.
Temperton
N J
NJ
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, UK.
Wilkinson
S R
SR
Meyer
D J
DJ
Kelly
J M
JM
D000818:Animals; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D017125:Genes, Protozoan; D005840:Gentian Violet / Q000494:pharmacology*; D009593:Nitroimidazoles / Q000494:pharmacology*; D010641:Phenotype; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics
D009593:Nitroimidazoles; D014344:Trypanocidal Agents; D013482:Superoxide Dismutase; D005840:Gentian Violet; C009999:benzonidazole