A 32-Kilodalton Hydrolase Plays an Important Role in Paracoccidioides brasiliensis Adherence to Host Cells and Influences Pathogenicity

Abstract

One of the most crucial events during infection with the dimorphic fungus

Decreased expression of 14-3-3 inParacoccidioides brasiliensisconfirms its involvement in fungal pathogenesis

“…Pb18 yeasts transformed with an empty vector (PbEV) were used as a Pb18 recombination control as previously described. 24,44 After transformation, the cells were spread onto selective BHI media supplemented with 200 mg mL ¡1 hygromycin B (Sigma-Aldrich). Prior to incubation, the plates were airdried in a safety cabinet for 30 min.…”

Lung Diseases - Selected State of the Art Reviews

“…In addition, P. brasiliensis is known to express adhesin-like molecules on its surface that recognize and bind to ECM proteins (Barbosa et al, 2006;González et al, 2005a). More recently, in elegant experiments using antisense technology, the role of a P. brasiliensisadhesin molecule was confirmed when a 32-kDa protein, a putative adhesion member of the haloacid dehalogenase (HAD) superfamily of hydrolases, conferred both adherence capacity to pulmonary human epithelial cells and virulence in a mouse model of infection (Hernández et al, 2010) The observations described above may suggest that ECM proteins modulate the pathogenesis of PCM by means of two mechanisms: 1) participation in the migration of proinflammatory cells into the lungs and 2) serving as a binding protein for P. brasiliensis expressing on its surface adhesin-like molecules (receptors). These interactions may serve the fungus both for attachment to host tissue (evading the physical barriers) and dissemination to other organs, contributing in this manner to the establishment of the disease process.…”

RNAi technology targeting Pb<i>GP</i>43 and Pb<i>P</i>27 in <i>Paracoccidioides brasiliensis</i>

“…However further studies need to be conducted for the synthesis of new and more stable constructions based on RNAi technology. Presently, use of the transformation system mediated by A. tumefaciens, which could show best results in P. brasiliensis gene silencing [23][24][25][26]. This is the first report using electrotransformation of hairpin construction based on RNAi technology targeting two important P. brasiliensis antigens (gp43 and p27).…”

“…Currently, a gene silencing strategy based on antisense-RNA (asRNA) has been efficiently employed in P. brasiliensis to induce the knockdown of PbCDC42, PbHAD32, PbAOX, and PbHSP90 [23][24][25][26]. Additionally, based in our previous results using the antisense RNA strategy targeting PbGP43 and PbP27 (unpublished), and the results obtained in this work using RNA interference strategy, we conclude that the non-stability in gene silencing is due to strategy and transformation methodology employed to down regulate specific gene.…”