Please use this identifier to cite or link to this item: http://www.arca.fiocruz.br/handle/icict/3730
Title: 1,3-β-d-Glucan synthase of Paracoccidioides brasiliensis: recombinant protein, expression and cytolocalization in the yeast and mycelium phases
Authors: Tomazett, Patrícia Kott
Félix, Carlos Roberto
Lenzi, Henrique Leonel
Faria, Fabrícia de Paula
Soares, Célia Maria de Almeida
Pereira, Maristela
Affilliation: Universidade Federal de Goiás. Instituto de Ciências Biológicas. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular. Goiânia, GO, Brasil
Universidade de Brasília. Instituto de Biologia. Laboratório de Enzimologia. Brasília, DF, Brasil
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Patologia. Rio de Janeiro, RJ, Brasil
Universidade Federal de Goiás. Instituto de Ciências Biológicas. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular. Goiânia, GO, Brasil
Universidade Federal de Goiás. Instituto de Ciências Biológicas. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular. Goiânia, GO, Brasil
Universidade Federal de Goiás. Instituto de Ciências Biológicas. Departamento de Bioquímica e Biologia Molecular. Laboratório de Biologia Molecular. Goiânia, GO, Brasil
Abstract: Paracoccidioides brasiliensis is a thermo-dimorphic human pathogenic fungus that in the mycelium phase lives at 23 C in environment and in the yeast phase at 37 C in the host tissues. In P. brasiliensis, the main polymers that compound the cell wall are chitin, 1,3-b-Dglucan and 1,3-a-glucan. They make a primary barrier responsible for the structural integrity and form of the cell wall. In P. brasiliensis, just one homologue of 1,3-b-D-glucan synthase gene (PbFKS1) was found. Here, the active recombinant protein (PbFks1pc) containing the catalytic region was obtained in Escherichia coli. In addition, a paradoxical dissociation was detected between the expression of the PbFKS1 transcript and the level of the corresponding protein PbFks1p, which was higher in the yeast phase, versus the amount of 1,3-b-D-glucan polymer, which was higher in the mycelium phase. Western blot analysis using protein extracts of cellular fractions showed that PbFks1p is present in the membrane-enriched fraction of mycelium and yeast cells and in the cell wall-enriched fractions of yeast cells. Confocal-immunocytolocalization of PbFks1p identified the protein in the apical growing region of the mycelium and distributed on the surface of the yeast cell. Two possible mechanisms could explain the above-mentioned discrepancy between the data: (a) overexpression of Rho1 GTPase as a regulator of 1,3-b-D-glucan synthase; (b) possible post-translational regulation of PbFks1p in P. brasiliensis isolates.
Keywords: Cell wall
Cytolocalization
1,3-b-D-glucan
Synthase
Paracoccidioides brasiliensis
Recombinant protein
DeCS: Paracoccidioides brasiliensis
Proteínas Recombinantes
Parede Celular
Issue Date: 2010
Citation: TOMAZETT, P. K. et al. 1,3-β-d-Glucan synthase of Paracoccidioides brasiliensis: recombinant protein, expression and cytolocalization in the yeast and myceliumphases. Fungal Biology, v. 114, p. 809-816, 2010.
DOI: http://dx.doi.org/10.1016/j.funbio.2010.07.007
ISSN: 1878-6146
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

Files in This Item:
File Description SizeFormat 
1,3-β-d-Glucan synthase of Paracoccidioides brasiliensi.pdf383.57 kBAdobe PDFThumbnail
    Request a copy


FacebookTwitterDeliciousLinkedInGoogle BookmarksBibTex Format mendeley Endnote DiggMySpace

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.