UNUSUAL T. cruzi ADENYLATE KINASE ISOFORM LOCATED IN THE CELL NUCLEUS

CAMARA, MARIA DE LOS MILAGROS; BOUVIER, LEÓN ALBERTO; GIACOMETTI, ALINA; MIRANDA, MARIANA; SAYÉ, MELISA; PEREIRA, CLAUDIO

Puerto Madryn

Encuentro; XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010

Cellular energetic homeostasis is partially maintained by the enzymes adenylate kinases (Adks) and nucleoside diphosphate kinases (NDPKs), which are involved in nucleotide interconvertion. In typical eukaryotic cells, there are very few isoforms of Adks and NDPKs, however, in trypanosomatids there have been characterized a large number of isoforms. This unusual number could be explained by the compartmentalized metabolism, which characterizes these organisms. In this work we studied a nuclear Adk isoform (Adkn) of T. cruzi. We measured the biochemical activity of this enzyme and generated transgenic parasites which express the protein fused to GFP to determine its subcellular localization. We also studied the Adkn trafficking between cytoplasm and nucleus under different drugs or media treatments, and also along the parasites growth curve. Furthermore, we mapped the non-canonical nuclear localization signal, by segmenting the protein and making fusions to GFP. The identification of the factors that regulate the subcellular localization and expression of this group of enzymes could be a powerful resource in the understanding cellular phosphotransfer network.