A quantitative view of the morphological phases of Paracoccidioides brasiliensis using proteomics

• Published in: Journal of proteomics 2011-12, Vol.75 (2), p.572-587
• Main Authors: Rezende, Tereza C.V., Borges, Clayton L., Magalhães, Adriana D., de Sousa, Marcelo Valle, Ricart, Carlos A.O., Bailão, Alexandre M., Soares, Célia M.A.
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; conidia; Diverse techniques; Fundamental and applied biological sciences. Psychology; Gel electrophoresis; gene expression regulation; Infection; mass spectrometry; Mass spectroscopy; Molecular and cellular biology; Morphogenesis; Morphological transition; Mycelia; mycelium; Paracoccidioides brasiliensis; Pathogens; Polymerase chain reaction; proteins; proteome; Proteomic analysis; proteomics; reverse transcriptase polymerase chain reaction; Transcription; two-dimensional gel electrophoresis; Virulence; Western blotting; yeasts
• ISSN: 1874-3919
• DOI: 10.1016/j.jprot.2011.08.020

Paracoccidioides brasiliensis is a fungal pathogen with a broad distribution in Latin American countries. The mycelia-to-yeast morphological transition of P. brasiliensis is involved in the virulence of this pathogen, and this event is essential to the establishment of infection. Here, we report the first proteomic comparison between the mycelia, the mycelia-to-yeast transition and the yeast cells. Changes in the relative abundance of the components of the proteome during phase conversion of P. brasiliensis were analyzed by two-dimensional gel electrophoresis coupled to mass spectrometry. Using MALDI-TOF-MS, we identified 100 total proteins/isoforms. We show that 18, 30 and 33 proteins/isoforms in our map are overexpressed in the mycelia, the mycelia-to-yeast transition and in yeast cells, respectively. Nineteen proteins/isoforms did not present significant differences in the volume spots in the three analyzed conditions. The differential expression was confirmed for six different proteins by Western blot analysis. The quantitative differences observed by the proteomic analysis were correlated with the transcript levels, as determined by quantitative RT-PCR of the analyzed conditions, including conidial formation and the transition from conidia-to-yeast cells. The analysis of the functional categories to which these proteins belong provided an integrated view of the metabolic reorganization during the morphogenesis of P. brasiliensis. [Display omitted] ► The first proteomic analysis of Paracoccidioides brasiliensis dimorphism. ► The transition to the yeast parasitic phase involves a shift in the metabolism. ► Proteins are differentially accumulated in the early transition process. ► Post translational modifications of proteins conform to different isoforms.