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UNVEILING THE TRYPANOSOMA CRUZI NUCLEAR PROTEOME
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Affilliation
Abstract
Replication of Trypanosoma cruzi, the etiological agent of Chagas disease, displays peculiar
features, such as absence of chromosome condensation and closed mitosis. Although
previous proteome and subproteome analyses of T. cruzi have been carried out, the nuclear
subproteome of this protozoan has not been described. Here, we report, for the first time to
the best of our knowledge, the isolation and proteome analysis of T. cruzi nuclear fraction.
For that, T. cruzi epimastigote cells were lysed and subjected to cell fractionation using two
steps of sucrose density gradient centrifugation. The purity of the nuclear fraction was confirmed
by phase contrast and fluorescence microscopy. Liquid chromatography coupled to
tandem mass spectrometry (LC-MS/MS) allowed the identification of 864 proteins. Among
those, 272 proteins were annotated as putative uncharacterized, and 275 had not been previously
reported on global T. cruzi proteome analysis. Additionally, to support our enrichment
method, bioinformatics analysis in DAVID was carried out. It grouped the nuclear
proteins in 65 gene clusters, wherein the clusters with the highest enrichment scores harbor
members with chromatin organization and DNA binding functions.
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