Resumo

The Paracoccidioides genus comprises human fungal pathogens which infect nearly 10 million people in Latin America, causing a systemic mycoses. The genus comprises four phylogenetic lineages (S1, PS2, PS3 and Pb01-like). P.lutzii strain, Pb01, and P. brasiliensis strain Pb18, that belong to the Pb01-like and S1 phylogenetic groups, respectively, were used in this study. Nucleus is an organelle responsible for containing the DNA responsible to carry the cell information. Proteins are directly connected interacting with DNA. Nuclear proteomic studies are important due the fact that proteins can be involved in regulatory process, nucleosome biogenesis, among others. Enriched nuclear proteome fraction of Paracaccidioides Pb01, was obtained. Immunofluorescence and electron microscopy were also performed. The proteins were digested using trypsin and identified using nanoUPLC-MS/MS, a two-dimensional liquid chromatography tandem mass spectrometry approach. Furthermore, in silico analysis was performed to determine the nuclear localization of the predicted proteins from the genomes of Paracoccidioides, at (http://www.broadinstitute.org/). For this purpose, we used the WoLF PSORT (http://wolfpsort.sep.cbrc.jp/) and NucPred (http://www.sbc.su.se/~maccallr/nucpred/) as bioinformatic tools. We found that the Pb01 genome presented 39 and 15% of the total genes encoding proteins which were classified as nuclear using WoLF PSORT and NucPred bioinformatic tools, respectively. Proteomic analysis showed 15 and 2% of proteins that were also classified as nuclear using the same strategy. To our knowledge, this is the first description of the nuclear proteome of Paracoccidioides. Additional studies will be performed comparing the proteomes of members of the different phylogenetic groups in the genus Paracoccidoides.