title: Unveiling the Trypanosoma cruzi Nuclear Proteome.
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.
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Agenor de Castro Moreira
authors.initials: Ade C
authors.lastname: dos Santos Júnior
authors.affiliation: Laboratório Interdisciplinar de Pesquisas Médicas, Instituto Oswaldo Cruz (IOC), Fundação Oswaldo Cruz (FIOCRUZ), 21040-360 Manguinhos, Rio de Janeiro, RJ, Brasil.
authors.forename: Dário Eluan
authors.initials: DE
authors.lastname: Kalume
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Camargo
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Diana Paola
authors.initials: DP
authors.lastname: Gómez-Mendoza
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: José Raimundo
authors.initials: JR
authors.lastname: Correa
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Charneau
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Marcelo Valle
authors.initials: MV
authors.lastname: de Sousa
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Beatriz Dolabela
authors.initials: BD
authors.lastname: de Lima
authors.affiliation: Department of Cell Biology, Institute of Biology, University of Brasilia, Campus Darcy Ribeiro, Asa Norte, 70910-900, Brasília, Brazil.
authors.forename: Carlos André Ornelas
authors.initials: CA
authors.lastname: Ricart
mesh_terms: D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D053719:Tandem Mass Spectrometry; D014349:Trypanosoma cruzi / Q000235:genetics / Q000378:metabolism*
chemical_list: D020543:Proteome; D015800:Protozoan Proteins