Unraveling the role of Aurora kinases in Trypanosoma cruzi

FASSOLARI, M.; FLAWIÁ, M.M.; TORRES, H.N.; ALONSO, G.D.

Villa Carlos Paz. Cordoba, Argentina.

Congreso; XLIV Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB).; 2008

Cell division and differentiation are key processes for protozoan pathogens to colonize and cause disease. Trypanosoma cruzi, the etiological agent of American Trypanosomiasis, possesses a complex life cycle that required strict checkpoints. Currently the regulation of cell division in T. cruzi is poorly understood. Aurora kinases (AUKs) are a family of conserved S/T kinases that are essential regulators of many events during cell division. Previously, we reported the identification of three AUK orthologs (TcAUK1, 2 and 3) in T. cruzi and expressed them in E. coli. In the present work, to investigate the AUK1 role in T. cruzi, we used pTREX and pTEX-GFP vectors to overexpress this enzyme either as a native protein or fused to the green fluorescent protein (GFP). As an alternative strategy to determinate the subcellular localization of this protein, we are using polyclonal antibodies against TcAUK1 in immunofluorescense assays. Considering that we failed to measure kinase activity of recombinant TcAUKs when we used commercial histones mix as substrate and the fact that T. cruzi histones show high divergence compared to other eukaryote ones, we decided to clone and express histones H3, H3 variant and H4 from T. cruzi in bacterial systems to use them as substrates in kinase activity assays. These results are a new step toward the clarification of TcAUKs role in the parasite life cycle.