Proteome characterization of Paracoccidioides lutzii conidia by using nanoUPLC-MSE

Moreira, André; Cruz-Leite, Vanessa Rafaela Milhomem; Silva, Lana O’Hara Souza; Parente, Ana Flávia Alves; Bailão, Alexandre Melo; Soares, Célia Maria de Almeida; Parente-Rocha, Juliana Alves; Hernández, Orville; Borges, Clayton Luiz

doi:10.1016/j.funbio.2020.05.004

Cited by 5 publications

(3 citation statements)

References 120 publications

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“…The formamidase gene of A. nidulans is regulated by NCR and presents a GATA motif in the promoter region [80]. The role of formamidase in the virulence of P. brasiliensis is under intensive investigation due to its high prevalence in the dimorphic phases of fungi, as shown by in vitro and ex vivo omics experiments [46,51]. Due to its association with determinants of virulence and nitrogen metabolism in Paracoccidioides spp., Silva et al [81] employed antisense RNA technology to silence the formamidase gene in P. lutzii.…”

Section: Discussionmentioning

confidence: 99%

“…Therefore, we chose this incubation time for further comparative proteomic analysis. NanoUPLC-MS E was conducted, following a previously outlined methodology [51], and the protein and peptide data generated were processed using PLGS, as depicted in Figure S2. Under NCR-proline conditions, 42.3% of the peptides were obtained from peptide match type data in the first pass, and 10.4% in the second pass.…”

Section: Proteomic Data Quality Analysismentioning

confidence: 99%

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Proteomics of Paracoccidioides lutzii: Overview of Changes Triggered by Nitrogen Catabolite Repression

Cruz-Leite,

Moreira,

Silva

et al. 2023

JoF

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Members of the Paracoccidioides complex are the causative agents of Paracoccidioidomycosis (PCM), a human systemic mycosis endemic in Latin America. Upon initial contact with the host, the pathogen needs to uptake micronutrients. Nitrogen is an essential source for biosynthetic pathways. Adaptation to nutritional stress is a key feature of fungi in host tissues. Fungi utilize nitrogen sources through Nitrogen Catabolite Repression (NCR). NCR ensures the scavenging, uptake and catabolism of alternative nitrogen sources, when preferential ones, such as glutamine or ammonium, are unavailable. The NanoUPLC-MSE proteomic approach was used to investigate the NCR response of Paracoccidioides lutzii after growth on proline or glutamine as a nitrogen source. A total of 338 differentially expressed proteins were identified. P. lutzii demonstrated that gluconeogenesis, β-oxidation, glyoxylate cycle, adhesin-like proteins, stress response and cell wall remodeling were triggered in NCR-proline conditions. In addition, within macrophages, yeast cells trained under NCR-proline conditions showed an increased ability to survive. In general, this study allows a comprehensive understanding of the NCR response employed by the fungus to overcome nutritional starvation, which in the human host is represented by nutritional immunity. In turn, the pathogen requires rapid adaptation to the changing microenvironment induced by macrophages to achieve successful infection.

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Section: Discussionmentioning

confidence: 99%

Section: Proteomic Data Quality Analysismentioning

confidence: 99%

Proteomics of Paracoccidioides lutzii: Overview of Changes Triggered by Nitrogen Catabolite Repression

Cruz-Leite,

Moreira,

Silva

et al. 2023

JoF

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“…Corroborating this, formamidase was identified as an exoantigen of P. lutzii [ 23 ], and was upregulated during ex vivo [ 24 , 25 ] and in vivo [ 26 ] experimental infection with P. brasiliensis. Additionally, formamidase was the most abundant on proteome analysis of P. lutzii conidia [ 27 ] and was upregulated during P. brasiliensis mycelium-to-yeast transition [ 28 ], suggesting a role of this enzyme during the first steps of infection with the Paracoccidioides species.…”

Section: Introductionmentioning

confidence: 99%

Paracoccidioides lutzii Formamidase Contributes to Fungal Survival in Macrophages

et al. 2022

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Nitrogen is a crucial nutrient for microorganisms that compose essential biomolecules. However, hosts limit this nutrient as a strategy to counter infections, therefore, pathogens use adaptive mechanisms to uptake nitrogen from alternative sources. In fungi, nitrogen catabolite repression (NCR) activates transcription factors to acquire nitrogen from alternative sources when preferential sources are absent. Formamidase has been related to nitrogen depletion in Aspergillus nidulans through formamide degradation to use the released ammonia as a nitrogen source. In Paracoccidioides spp., formamidase is highly expressed in transcriptomic and proteomic analyses. Here, we aim to investigate the importance of formamidase to Paracoccidioides lutzii. Thereby, we developed a P. lutzii silenced strain of fmd gene (AsFmd) by antisense RNA technology using Agrobacterium tumefaciens-mediated transformation (ATMT). The AsFmd strain led to increased urease expression, an enzyme related to nitrogen assimilation in other fungi, suggesting that P. lutzii might explore urease as an alternative route for ammonia metabolism as a nitrogen source. Moreover, formamidase was important for fungal survival inside macrophages, as fungal recovery after macrophage infection was lower in AsFmd compared to wild-type (WT) strain. Our findings suggest potential alternatives of nitrogen acquisition regulation in P. lutzii, evidencing formamidase influence in fungal virulence.

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Insight into the metabolic changes during germination of Aspergillus niger conidia using nLC-qTOF

Shankar

2022

Biologia

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Proteome characterization of Paracoccidioides lutzii conidia by using nanoUPLC-MSE

Cited by 5 publications

References 120 publications

Proteomics of Paracoccidioides lutzii: Overview of Changes Triggered by Nitrogen Catabolite Repression

Proteomics of Paracoccidioides lutzii: Overview of Changes Triggered by Nitrogen Catabolite Repression

Paracoccidioides lutzii Formamidase Contributes to Fungal Survival in Macrophages

Insight into the metabolic changes during germination of Aspergillus niger conidia using nLC-qTOF

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