Role of the Trypanosoma cruzi Pantothenate transporter (TcPPT1) in survival under stress conditions

D, RUIZ; L. FRACCAROLI,; BALCAZAR, D; L. LAROCCA; TORRES, PS; CARRILLO C

Congreso; XXIX Reunión de la Sociedad Argentina de Protozoología, en el marco de la Reunión Conjunta de Sociedades de Biociencias; 2017

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 based on their poor specificity. Thus, there is a need to identify new targets to develop novel tripanocidal strategies.Vitamins are essential micronutrients for all living cells; in particular B vitamins are relevant in the biology of T. cruzi. Previously, we have shown that pantothenate, the precursor of Coenzyme A, has a key role in T. cruzi metabolism. In silico studies suggested that T. cruzi is auxotrophous for this vitamin, and we identified a putative pantothenate transporter (TcPPT1). The aim of this work is to study TcPPT1 role in T. cruzi proliferation under stress conditions.To approach this aim, we constructed mutant Y-GFP strain epimastigotes that overexpress TcPPT1 fused to a mCherry tag. Its presence and expression were assessed by PCR, RT-PCR and fluorescence microscopy.To evaluate possible differential proliferative responses we performed cultures of wild type (mCherry) and transgenic (TcPPT1-mCherry) epimastigotes under different stressing conditions: nutrient deprivation (20nM pantothenate), oxidative stress (H2O2 0-150uM) and tripanocidal drugs (Bnz and Nfx 0-100uM). Proliferation rate of TcPPT1-mCherry epimastigotes was significantly higher than mCherry epimastigotes in low pantothenate conditions (p