Localization and Enzymatic Activity of Aurora Kinases from Trypanosoma cruzi

FASSOLARI M; MEYER CG; TORRES HN; FLAWIÁ MM; ALONSO GD

Puerto Madryn, Chubut.

Congreso; XLVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2010

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB)

In all eukaryotes protein kinases are crucial players in the mechanism of cell cycle regulation. Among them are the Aurora kinases controlling important processes during mitosis. This family of enzymes was characterized in several organisms, from protozoan to mammals. One of the most interesting protozoan families are the trypanosomatids because of their complex life cycle, here are included T. cruzi, T. brucei and Leishmania major. Aurora kinases were identified in L. major and were deeply studied in T. brucei (TbAUK1). As in mammals, TbAUK1 is involved in mitotic events like spindle formation and dynamics, chromatin segregation and cytokinesis. In T. cruzi we?ve already identified three genes that codify for aurora kinases (TcAUK1, 2 and 3) and have described their genomic organization and expression profile at mRNA level. We have also established TcAUK1 as a nuclear protein in epimastigotes that its expression seems to be regulated in a cell cycle-dependent way. In this work we extend the study of cellular localization to the others TcAUKs, expressing them as fusion proteins with GFP in epimastigotes of T. cruzi. As part of the biochemical characterization of these enzymes we performed kinase assays, using recombinant TcAUKs expressed in E. coli. With these assays we could established that TcAUKs show greater activity levels when Mn2+ is used as cofactor.