The epigenetic modulator BDF2 is involved in the normal progression of Trypanosoma cruzi life cycle.

PEZZA A; SERRA EC; ALONSO VL; TAVERNELLI LE

Mar del Plata

Congreso; Reunion Anual de Sociedades de Biociencias SAIC -SAFE - SAB - SAP; 2019

Sociedad Argentina de Protozoologia

Trypansoma cruzi is the causative agent of Chagas? disease. In this parasite, protein-coding genes are transcribed polycistronically without canonical promoters or typical gene transcription initiation control. In this context, epigenetic regulation and post-trancriptional regulating mechanisms Bromodomains are protein interaction modules that selectively recognize and bind acetyl-lysine residues present in histone and non-histone proteins The bromodomain factor 2 of T. cruzi (TcBDF2) is a nuclear protein containing a bromodomain on its N-terminal portion. This domain enables the protein to interact whit acetylated lysine on chromatin proteins, like H4 histones, becoming an important factor on epigenetic regulation. Working with truncated mutant version of BDF2, we determine the nuclear localization signal of the protein by inmunofluorescent microscopy. In addition we constructed mutant versions of the protein and corroborates by in vitro assays that these punctual modifications impaired the ligand binding. We analyzed the inducible overexpression of the wild type and mutant proteins over the life cycle of the parasite determining that Bdf2 is an important factor for the normal growth of the epimastigotes. Moreover, this protein is involved on the metacyclic trypomastigote differentiation and is crucial for the tripomastigotes release from VERO cells once the infection is established. Finally, using a fluorescence quenching assay we determined the ability of BDF2 to bind to commercial bromodomain inhibitors. We determined the dissociation constant for bromosporine and I-BET151.