Paracoccidioides brasiliensis PbP27 gene: Knockdown procedures and functional characterization

Paracoccidioides brasiliensis PbP27 gene encodes a protein localized in both the fungal cytoplasm and cell wall. The parasitic infectious form produces this protein preferentially with the gene's expression varying between the fungus phylogenetic species. The biological function of the native p...

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Autores:
Tipo de recurso:
Fecha de publicación:
2014
Institución:
Universidad del Rosario
Repositorio:
Repositorio EdocUR - U. Rosario
Idioma:
eng
OAI Identifier:
oai:repository.urosario.edu.co:10336/24057
Acceso en línea:
https://doi.org/10.1111/1567-1364.12099
https://repository.urosario.edu.co/handle/10336/24057
Palabra clave:
Complementary RNA
Gamma interferon
HLA DR antigen
Oligonucleotide
Protein p27
Epitope
Fungal protein
Gamma interferon
HLA DR antigen
Peptide
Protein binding
Agrobacterium tumefaciens
Allele
Amino acid sequence
Article
Cell structure
Cell viability
Fungal gene
Fungal virulence
Fungus growth
Gene expression
Gene frequency
Macrophage
Mycelium
Nonhuman
Nucleotide sequence
Paracoccidioides brasiliensis
Pbp27 gene
Protein function
Real time polymerase chain reaction
T lymphocyte
Yeast cell
Chemistry
Drug effects
Epitope mapping
Gene expression regulation
Gene silencing
Genetics
Immunology
Metabolism
Microbiology
Paracoccidioides
Agrobacterium tumefaciens
Fungi
Paracoccidioides brasiliensis
Amino Acid Sequence
Epitope Mapping
Fungal Proteins
Gene Knockdown Techniques
HLA-DR Antigens
Interferon-gamma
Macrophages
Paracoccidioides
Peptides
Protein Binding
Antisense RNA
Fungal survival
Gene silencing
P27
Paracoccidioides brasiliensis
Pbp27
Fungal
T-Lymphocyte
Epitopes
Gene Expression Regulation
Rights
License
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Description
Summary:Paracoccidioides brasiliensis PbP27 gene encodes a protein localized in both the fungal cytoplasm and cell wall. The parasitic infectious form produces this protein preferentially with the gene's expression varying between the fungus phylogenetic species. The biological function of the native p27 has yet to be determined during either growth of the yeast or host infection. Therefore, in this study, through the use of antisense RNA technology and Agrobacterium tumefaciens-mediated transformation, we generated mitotically stable PbP27 mutants (PbP27 aRNA) with the goal to evaluate the role of p27 in the biology and virulence of this fungus. PbP27 expression was reduced 60-75% in mutants, as determined by real-time PCR in correlation with a decrease in p27 expression. No alterations in the growth curve or in the ability to shift from mycelia to yeast or from yeast to mycelia were observed in PbP27 aRNA strains; however, we did observe a reduction in cell vitality. Moreover, a decrease in cell viability of PbP27 aRNA yeast cells after interaction with IFN-?-stimulated macrophages was detected. Based on these results, we propose that p27 plays a role in yeast cell architecture and represents one of the mechanisms employed by this fungus for its interaction with the monocyte/macrophage system. p27 plays a role in Paracoccidioides brasiliensis yeast cell architecture and represents one of the mechanisms employed by the fungus for its interaction with the monocyte/macrophage system. © 2013 Federation of European Microbiological Societies. Published by John Wiley and Sons Ltd. All rights reserved.