Effect of Nitrosative Stress on theS-Nitroso-Proteome ofParacoccidioides brasiliensis

Author(s)

Navarro, Marina V. ^[1] ; Chaves, Alison F. A. ^[1] ; Castilho, Daniele G. ^[1] ; Casula, Isis ^[2] ; Calado, Juliana C. P. ^[1] ; Conceicao, Palloma M. ^[2] ; Iwai, Leo K. ^[3] ; de Castro, Beatriz F. ^[2] ; Batista, Wagner L. ^{[1, 2]}

Affiliation

^[1] Univ Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Escola Paulista Med, Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, Dept Pharmaceut Sci, Inst Ciencias Ambientais Quim & Farmaceut, Diadema - Brazil

^[3] Inst Butantan, Lab Appl Toxinol, Ctr Toxins Immune Response & Cell Signaling, Sao Paulo - Brazil

• Document type: Journal article
• Source: FRONTIERS IN MICROBIOLOGY; v. 11, JUN 4 2020.
• Web of Science Citations: 0

Abstract

The fungiParacoccidioides brasiliensisandParacoccidioides lutziiare the causative agents of paracoccidioidomycosis (PCM), a systemic mycosis endemic to Latin America. This fungus is considered a facultative intracellular pathogen that is able to survive and replicate inside macrophages. The survival of the fungus during infection depends on its adaptability to various conditions, such as nitrosative/oxidative stress produced by the host immune cells, particularly alveolar macrophages. Currently, there is little knowledge about theParacoccidioidesspp. signaling pathways involved in the fungus evasion mechanism of the host defense response. However, it is known that some of these pathways are triggered by reactive oxygen species and reactive nitrogen species (ROS/RNS) produced by host cells. Considering that the effects of NO (nitric oxide) on pathogens are concentration dependent, such effects could alter the redox state of cysteine residues by influencing (activating or inhibiting) a variety of protein functions, notablyS-nitrosylation, a highly important NO-dependent posttranslational modification that regulates cellular functions and signaling pathways. It has been demonstrated by our group thatP. brasiliensisyeast cells proliferate when exposed to low NO concentrations. Thus, this work investigated the modulation profile ofS-nitrosylated proteins ofP. brasiliensis, as well as identifyingS-nitrosylation sites after treatment with RNS. Through mass spectrometry analysis (LC-MS/MS) and label-free quantification, it was possible to identify 474 proteins in theS-nitrosylated proteome study. With this approach, we observed that proteins treated with NO at low concentrations presented a proliferative response pattern, with several proteins involved in cellular cycle regulation and growth being activated. These proteins appear to play important roles in fungal virulence. On the other hand, fungus stimulated by high NO concentrations exhibited a survival response pattern. Among theseS-nitrosylated proteins we identified several potential molecular targets for fungal disease therapy, including cell wall integrity (CWI) pathway, amino acid and folic acid metabolisms. In addition, we detected that the transnitrosylation/denitrosylation redox signaling are preserved in this fungus. Finally, this work may help to uncover the beneficial and antifungal properties of NO in theP. brasiliensisand point to useful targets for the development of antifungal drugs. (AU)

• FAPESP's process: 17/04592-0 - Evaluation of S-nitrosylated proteins of the Paracoccidioides brasiliensis after nitrosative stress and identification of potential redox signaling pathways
• Grantee: Wagner Luiz Batista
• Support Opportunities: Regular Research Grants
• FAPESP's process: 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling
• Grantee: Hugo Aguirre Armelin
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 16/04000-3 - Pharmacology of oligopeptidases and intracellular peptides
• Grantee: Emer Suavinho Ferro
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 15/09654-9 - Global analysis of S-nitrosylation sites in proteins of Paracoccidioides brasiliensis after nitrosative stress
• Grantee: Marina Valente Navarro
• Support Opportunities: Scholarships in Brazil - Master
• FAPESP's process: 17/17943-6 - Proteomic and peptidomic analysis of snake venom effect on cancer cell lines and mouse brain tissues
• Grantee: Leo Kei Iwai
• Support Opportunities: Regular Research Grants