Alterations of protein expression in conditions of copper-deprivation for Paracoccidioides lutzii in the presence of extracellular matrix components

Abstract

BackgroundParacoccidioides spp is a fungi genus and the agent of paracoccidioidomycosis. The strategies of infection used by these pathogens involve the expression of proteins related to adaptation to the host, particularly regarding the uptake of micronutrients. This study analyzed the adhesion of Paracoccidioides lutzii during conditions of copper (Cu) and iron (Fe) deprivation, while also evaluating the proteins expressed in conditions of Cu depletion in the presence of four extracellular matrix (ECM) components (laminin, fibronectin and types I and IV collagen).ResultsWe cultured the P. lutzii in a chemically defined media without Cu and Fe. The fungus was then placed in contact with different ECM components and adhesion was evaluated. A significant increase in binding to all ECM components was observed when the fungus was cultured without Cu; which might be related to some adhesins expression. A proteomic assay was developed and revealed 39 proteins expressed that are involved in processes such as virulence, protein synthesis, metabolism, energy, transcription, transport, stress response and the cell cycle when the fungus was interacting with the ECM components. The up-regulated expression of two important adhesins, enolase and 14-3-3, was observed at the fungal cell wall during the interaction with the ECM components, indicating the role of these proteins in the Paracoccidioides–host interaction.ConclusionsThis study is important for determining prospective proteins that may be involved in the interaction of Paracoccidioides with a host. Understanding the adaptive response to different growth conditions, elucidating the processes of adhesion and cell invasion, and identifying the proteins that are differentially expressed during the fungus-host interaction may help elucidate mechanisms used for survival and growth of Paracoccidioides in various human tissues.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-014-0302-7) contains supplementary material, which is available to authorized users.

Interactome of Glyceraldehyde-3-Phosphate Dehydrogenase Points to the Existence of Metabolons in Paracoccidioides lutzii

“…Glyceraldehyde-3-phosphate dehydrogenase is a moonlight protein with a multitude of functions (Barbosa et al, 2006; Rezende et al, 2011; Weber et al, 2012; de Oliveira et al, 2013, 2014; Parente et al, 2013) due to interactions with other proteins. Here, GAPDH interacts with several proteins related to energy metabolism via glycolysis, TCA, respiration, glyoxylate and methylcitrate pathways.…”

“…GAPDH has been intensely investigated in Paracoccidioides . It is differentially expressed in several approaches such as copper and zinc deprivation in P. lutzii (Parente et al, 2013; de Oliveira et al, 2014), up regulated in P. brasiliensis yeast cells (Rezende et al, 2011), acting as adhesin (Barbosa et al, 2006), present in the secretome of mycelia and yeast cells (Weber et al, 2012) and interacting with malate synthase (de Oliveira et al, 2013). As a moonlight macromolecule, GAPDH interacts with a large variety of proteins, forming multiprotein complexes that play unique roles within cells (Brandina et al, 2006; Ferreira et al, 2013).…”

Metabolic Adaptation of Paracoccidioides brasiliensis in Response to in vitro Copper Deprivation

“…These important processes are related to detoxification (Peña et al, 1998), respiration (Horng et al, 2004) and capture of metals (Georgatsou et al, 1997). Therefore, the absence of copper may compromise the fungus' ability to adapt, survive and infect the host (Ding et al, 2014b;De Oliveira et al, 2014;Broxton and Culotta, 2016). Through a proteomic approach, we identified differentially accumulated proteins related to beta oxidation, glyoxylate cycle, pentose phosphate pathway and cell wall polymer synthesis, as well as others compromised with cellular respiration and detoxification.…”

Involvement of Dihydrolipoyl Dehydrogenase in the Phagocytosis and Killing of Paracoccidioides brasiliensis by Macrophages

“…infection is a survival strategy (Lima et al, 2014 ). P. lutzii , when grown under copper deprivation, showed induction of the pyruvate dehydrogenase complex, suggesting that availability of acetyl-CoA by the generated pyruvate could be important for the survival of P. lutzii in inhospitable conditions (de Oliveira et al, 2014 ). Parente-Rocha et al ( 2015 ) demonstrated that the fungi could modify their metabolism when subjected to conditions that mimic macrophage infection by P. brasiliensis .…”