Employing proteomic analysis to compare Paracoccidioides lutzii yeast and mycelium cell wall proteins

Araújo, Danielle Silva; Lima, Patrícia de Sousa; Baeza, Lílian Cristiane; Parente, Ana Flávia Alves; Bailão, Alexandre Melo; Borges, Clayton Luiz; Soares, Célia Maria de Almeida

doi:10.1016/j.bbapap.2017.08.016

Cited by 18 publications

(12 citation statements)

References 95 publications

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“…Ecm33 is a GPI-linked cell wall protein that plays an important role for cell wall integrity and architecture in C. albicans (Martinez-Lopez et al, 2004, 2006). In P. lutzii , this protein occurs in the mycelium cell wall, in agreement to the data here presented (Araújo et al, 2017).…”

Section: Discussionsupporting

confidence: 92%

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

et al. 2019

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Paracoccidioidomycosis (PCM), a systemic mycosis with a high incidence in Latin America, is caused by thermodimorphic fungi of the Paracoccidioides genus. The contact with host occurs by the inhalation of conidia or mycelial propagules which once reaching the pulmonary alveoli differentiate into yeast cells. This transition process is vital in the pathogenesis of PCM allowing the fungus survival in the host. Thus, the present work performed a comparative proteome analysis of mycelia, mycelia-to-yeast transition, and yeast cells of Paracoccidioides brasiliensis . For that, tryptic peptides were labeled with iTRAQ and identified by LC–MS/MS and computational data analysis, which allowed the identification of 312 proteins differentially expressed in different morphological stages. Data showed that P. brasiliensis yeast cells preferentially employ aerobic beta-oxidation and the tricarboxylic acid cycle accompanied by oxidative phosphorylation for ATP production, in comparison to mycelia and the transition from mycelia-to-yeast cells. Furthermore, yeast cells show a metabolic reprogramming in amino acid metabolism and in the induction of virulence determinants and heat shock proteins allowing adaptation to environmental conditions during the increase of the temperature. In opposite of that, the alcoholic fermentation found to P. lutzii , at least under laboratory conditions, is strongly favored in mycelium compared to yeast cells. Thereby, the data strongly support substantial metabolic differences among members of the Paracoccidioides complex, when comparing the saprobiotic mycelia and the yeast parasitic phases.

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Section: Discussionsupporting

confidence: 92%

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

et al. 2019

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“…Although intriguing, our results are in accordance with findings in the literature. Approaches of the cell wall proteomes of P. brasiliensis and C. albicans also evidenced the presence of classic cytoplasmic proteins in the cell wall [28,55,56]. Some authors argue that the presence of these proteins in the cell wall could be due to the methodology used to obtain the protein extracts.…”

Section: Discussionmentioning

confidence: 99%

“…To obtain P. lutzii CWPs, we employed the methodology described by Araújo et al, (2017) [28], with some modifications. After 48 h of fungal culture in the presence of hemoglobin (treatment) or in iron deprivation (control), the cultures were collected and centrifuged at 800× g for 10 min at 4 • C, and cells were washed 5 times with lysis buffer (10 mM Tris-HCl pH 8.5, 2 mM CaCl 2 , 1:10 protease inhibitor).…”

Section: Extraction Of P Lutzii Cell Wall Proteins (Cwps)mentioning

confidence: 99%

“…The authors obtained F1 cell wall enriched extracts and identified proteins, such as serine proteinase and fructose-1,6-bisphosphate aldolase interacting with macrophage surface proteins, and confirmatory assays showed that these proteins undergo positive regulation during the interaction of the fungus with the host [34]. The studies performed by Araújo et al (2017) [28] and Tomazett et al (2019) [34] demonstrated that the identification of the cell wall proteins is a good alternative to list new targets in the context of host-pathogen interaction.…”

Section: Introductionmentioning

confidence: 99%

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Interacting with Hemoglobin: Paracoccidioides spp. Recruits hsp30 on Its Cell Surface for Enhanced Ability to Use This Iron Source

Souza

Tomazett

Silva

et al. 2021

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Paracoccidioides spp. are thermally dimorphic fungi that cause paracoccidioidomycosis and can affect both immunocompetent and immunocompromised individuals. The infection can lead to moderate or severe illness and death. Paracoccidioides spp. undergo micronutrients deprivation within the host, including iron. To overcome such cellular stress, this genus of fungi responds in multiple ways, such as the utilization of hemoglobin. A glycosylphosphatidylinositol (GPI)-anchored fungal receptor, Rbt5, has the primary role of acquiring the essential nutrient iron from hemoglobin. Conversely, it is not clear if additional proteins participate in the process of using hemoglobin by the fungus. Therefore, in order to investigate changes in the proteomic level of P. lutzii cell wall, we deprived the fungus of iron and then treated those cells with hemoglobin. Deprived iron cells were used as control. Next, we performed cell wall fractionation and the obtained proteins were submitted to nanoUPLC-MSE. Protein expression levels of the cell wall F1 fraction of cells exposed to hemoglobin were compared with the protein expression of the cell wall F1 fraction of iron-deprived cells. Our results showed that P. lutzii exposure to hemoglobin increased the level of adhesins expression by the fungus, according to the proteomic data. We confirmed that the exposure of the fungus to hemoglobin increased its ability to adhere to macrophages by flow cytometry. In addition, we found that HSP30 of P. lutzii is a novel hemoglobin-binding protein and a possible heme oxygenase. In order to investigate the importance of HSP30 in the Paracoccidioides genus, we developed a Paracoccidioides brasiliensis knockdown strain of HSP30 via Agrobacterium tumefaciens-mediated transformation and demonstrated that silencing this gene decreases the ability of P. brasiliensis to use hemoglobin as a nutrient source. Additional studies are needed to establish HSP30 as a virulence factor, which can support the development of new therapeutic and/or diagnostic approaches.

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“…Accordingly, in P . lutzii proteomic studies showed that chitinase 3 was only detected in the cell wall of the mycelium (Araújo et al, 2017). Similarly, in Candida albicans the chitinase activity is higher in hypha and production, and hydrolysis of chitin was induced during yeast-hypha morphogenesis (Selvaggini et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

Cell Wall Synthesis, Development of Hyphae and Metabolic Pathways Are Processes Potentially Regulated by MicroRNAs Produced Between the Morphological Stages of Paracoccidioides brasiliensis

et al. 2018

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MicroRNAs are molecules involved in post-transcriptional gene regulation. In pathogenic fungi, microRNAs have been described at different morphological stages by regulating targets involved in processes such as morphogenesis and energy production. Members of the Paracoccidioides complex are the main etiological agents of a systemic mycosis in Latin America. Fungi of the Paracoccidioides complex present a wide range of plasticity to colonize different niches. In response to environmental changes these fungi undergo a morphological switch, remodel their cellular metabolism and modulate structural cell wall components. However, the underlying mechanisms regulating the gene expression is not well understood. By using high performance sequencing and bioinformatics analyses, this work characterizes microRNAs produced by Paracoccidioides brasiliensis. Here, we demonstrated that the transcript encoding proteins involved in microRNA biogenesis were differentially expressed in each morphological stage. In addition, 49 microRNAs were identified in cDNA libraries with 44 differentially regulated among the libraries. Sixteen microRNAs were differentially regulated in comparison to the mycelium in the mycelium-to-yeast transition phase. The yeast parasitic phase revealed a complete remodeling of the expression of these small RNAs. Analyses of targets of the induced microRNAs, from the different libraries, revealed that these molecules may potentially regulate in the cell wall, by repressing genes involved in the synthesis and degradation of glucans and chitin. Furthermore, mRNAs involved in cellular metabolism and development were predicted to be regulated by microRNAs. Therefore, this work describes a putative post transcriptional regulation, mediated by microRNAs in P. brasiliensis and its influence on the adaptive processes of thermal dimorphic fungus.

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Employing proteomic analysis to compare Paracoccidioides lutzii yeast and mycelium cell wall proteins

Cited by 18 publications

References 95 publications

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

Interacting with Hemoglobin: Paracoccidioides spp. Recruits hsp30 on Its Cell Surface for Enhanced Ability to Use This Iron Source

Cell Wall Synthesis, Development of Hyphae and Metabolic Pathways Are Processes Potentially Regulated by MicroRNAs Produced Between the Morphological Stages of Paracoccidioides brasiliensis

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