Differential PbP27 expression in the yeast and mycelial forms of the Paracoccidioides brasiliensis species complex

p27 is an antigenic protein produced by Paracoccidioides brasiliensis, the etiologic agent of paracoccidioidomycosis (PCM). Despite its unknown function, it has been suggested as a putative virulence factor, proposed as a suitable target for the design of diagnostic tools and vaccines, and considered as an enhancer in antifungal treatment of PCM. We evaluated sequence polymorphisms of PbP27 gene sequence among isolates, finding some polymorphisms associated with the isolates' phylogenetic origin. In order to determine if there was a differential expression pattern between morphological states and among isolates, we also evaluated PbP27 expression, at transcriptional and translational levels, in mycelia and yeast cultures in 14 isolates belonging to the P. brasiliensis species complex (S1, PS2, PS3, and 'Pb01-like', proposed to be named Paracoccidioides lutzii) by two techniques, real time RT-PCR (RT-qPCR) and protein dot blot. For the latter, four protein extracts from different cell localizations (SDS or ?-mercaptoethanol, cytoplasmic and extracellular proteins) were analyzed for each isolate. p27 was present in the four extracts evaluated, mainly in the SDS extract, corresponding to an extract containing proteins loosely attached to the cell wall. This information correlates with immunohistochemical analysis, where positive staining of the yeasts' cell wall was observed. We found that p27 was present in all isolates, mainly in the yeast form. This pattern was corroborated by RT-qPCR results, with higher expression levels found in the yeast form for most of the isolates. The results provide new insights into the expression patterns of this protein, and further characterize it in view of potential uses as a diagnostic and/or therapeutic tool. © 2011 Elsevier Inc.

• Fecha de publicación: 2011
• Institución: Universidad del Rosario
• Repositorio: Repositorio EdocUR - U. Rosario
• Idioma: eng
• OAI Identifier: oai:repository.urosario.edu.co:10336/24108
• Acceso en línea: https://doi.org/10.1016/j.fgb.2011.09.001; https://repository.urosario.edu.co/handle/10336/24108
• Palabra clave: Cytoplasm protein; Fungus antigen; Mercaptoethanol; Protein p27; Antigen expression; Article; Cell wall; Cellular distribution; Controlled study; Dna polymorphism; Dot hybridization; Fungal gene; Fungal genetics; Fungal phenomena and functions; Fungal strain; Fungus culture; Fungus isolate; Gene sequence; Genetic transcription; Immunohistochemistry; Mycelium; Nonhuman; Nucleotide sequence; Paracoccidioides; Paracoccidioides brasiliensis; Paracoccidioides brasiliensis species complex; Paracoccidioides brasiliensis type pb01 like; Paracoccidioides brasiliensis type ps2; Paracoccidioides brasiliensis type ps3; Paracoccidioides brasiliensis type s1; Paracoccidioides lutzii; Pbp27 gene; Phylogeny; Polyacrylamide gel electrophoresis; Priority journal; Protein localization; Real time polymerase chain reaction; Reverse transcription polymerase chain reaction; Rna translation; Sequence analysis; South american blastomycosis; Strain difference; Western blotting; Yeast; Cell wall; Fungal proteins; Immunohistochemistry; Paracoccidioides; Real-time polymerase chain reaction; Reverse transcriptase polymerase chain reaction; Yeasts; Paracoccidioides; Paracoccidioides brasiliensis; Dimorphism; Dot blot; P27 antigen; Paracoccidioides brasiliensis; Paracoccidioidomycosis; Qpcr; Western blot; polyacrylamide gel; genetic; fungal; fungal; dna; fungal; western; Antigens; Blotting; Electrophoresis; Gene expression regulation; Genes; Polymorphism; Sequence analysis
• License: Abierto (Texto Completo)