INVESTIGADORES
FRACCAROLI Laura Virginia
congresos y reuniones científicas
Título:
Identification and characterization of the Trypanosoma cruzi Pantothenate Transporter (TcPPT1) and its role in survival under stress conditions
Autor/es:
MD. RUIZ; L. FRACCAROLI; D. BALCAZAR; L. LAROCCA; P. TORRES; C. CARRILLO
Lugar:
Buenos Aires
Reunión:
Congreso; 18th International Congress on Infectious Diseases; 2018
Resumen:
Background: Chagas disease is an endemic parasitosis originally from Latin America, caused by the protozoan Trypanosoma cruzi. The current therapies are limited in efficacy and show multiple side effects. Thus, there is a need to identify new targets to develop novel, more effective and specific trypanocidalstrategies. Vitamins are essential micronutrients for all living cells; in particular, B vitamins are relevant in the biology of T. cruzi. Provided that pantothenate, the precursor of Coenzyme A, is involved in numerous essentialcellular reactions and that our previous in silico studies suggested that T. cruzi is auxotrophous for this vitamin, the aim of this work was to study pantothenate role in T. cruzi epimastigotes and to identify and characterize potential pantothenate-transporters.Methods & Materials: In order to approach this aim, we evaluated T. cruzi epimastigotes (Y-GFP strain) proliferation and viability performing growth curves and MTT assay in a culture media with low concentration of pantothenate. Provided that our bioinformatic studies yielded the identification of a putative pantothenate transporter (TcPPT1), we constructed a mutant Y-GFP strain that overexpress TcPPT1 fused to a mCherry tag. Then we evaluated differential proliferative response under different stressing conditions: nutrient deprivation (20nM pantothenate), oxidative stress (0-150uM H2O2) and trypanocidal drugs (0-100uM Benznidazole and Nifurtimox). Results: T. cruzi epimastigotes proliferation rate was significantly decreased in culture media with low concentration of pantothenate. This effect was reverted, in a concentration dependent manner, when the vitamin was added to the media. The pantothenate proliferative effect evidenced on T. cruzi epimastigotes was not observed in other axenic cultures of related trypanosomatids (T. brucei, L. mexicana and C. fasciculata). TcPPT1 overexpression was assessed by PCR, RT-PCR and fluorescence microscopy in mutant Y-GFP clones. Proliferation rate of TcPPT1-mCherry epimastigotes was significantly higher than mCherry epimastigotes (control) in low pantothenate conditions (p