Screening of molecules directed to the host cell nucleus in Trypanosoma cruzi

DURANTE I; CARMONA SJ; AGÜERO F; A BUSCAGLIA, CARLOS

San Luis

Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2011

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Trypanosoma cruzi (Tc), the protozoan parasite causative of Chagas` disease, secretes abundant molecules in all developmental stages of its complex life cycle. One intriguing possibility is that intracellular trypomastigotes (T) and amastigotes (A), secrete molecules to the host cell nucleus to co-opt cellular processes. In this work, an in silico strategy was developed to search for candidate molecules potentially directed to the host cell nucleus during a Tc Infection, using the bioinformatic tool TDRTargets. The genome of Tc was screened looking for molecules with a number of defined features: a) Presence of a Signal Peptide (SP), b), Consensus for the addition of GPI, c) Presence of Nuclear Localization Signals (NLS), e) SP-independent secretion, and d) Nuclear features, among others. After manual curation of results, some candidates were selected for biochemical characterization, involving: a), Evaluation of predicted NLS functionality, b) Study of expression and subcellular localization in Tc developmental forms. As a result of these studies, some interesting candidates emerged: a protein with homology to CEST (Chaperone for E. coli Secretion of Tir), a protein with a WD40 domain, homologue of CAF-1B (Chromatin Assembly Factor 1B), and a protein with unknown putative function. These candidates are being subjected to further biochemical characterization.