Aspergillus fumigatus High Osmolarity Glycerol Mitogen Activated Protein Kinases SakA and MpkC Physically Interact During Osmotic and Cell Wall Stresses

Author(s)

Manfiolli, Adriana Oliveira ^[1] ; Mattos, Eliciane Cevolani ^[1] ; de Assis, Leandro Jose ^[1] ; Silva, Lilian Pereira ^[1] ; Ulas, Mevlut ^[2] ; Brown, Neil Andrew ^[3] ; Silva-Rocha, Rafael ^[4] ; Bayram, Ozgur ^[2] ; Goldman, Gustavo H. ^[1]

Affiliation

^[1] Univ Sao Paulo, Fac Ciencias Farmaceut Ribeirao Preto, Ribeirao Preto - Brazil

^[2] Maynooth Univ, Dept Biol, Maynooth, Kildare - Ireland

^[3] Univ Bath, Dept Biol & Biochem, Bath, Avon - England

^[4] Univ Sao Paulo, Fac Med Ribeirao Preto, Ribeirao Preto - Brazil

• Document type: Journal article
• Source: FRONTIERS IN MICROBIOLOGY; v. 10, MAY 7 2019.
• Web of Science Citations: 1

Abstract

Aspergillus fumigatus, a saprophytic filamentous fungus, is a serious opportunistic pathogen of mammals and it is the primary causal agent of invasive aspergillosis (IA). Mitogen activated protein Kinases (MAPKs) are important components involved in diverse cellular processes in eukaryotes. A. fumigatus MpkC and SakA, the homologs of the Saccharomyces cerevisiae Hog1 are important to adaptations to oxidative and osmotic stresses, heat shock, cell wall damage, macrophage recognition, and full virulence. We performed protein pull-down experiments aiming to identify interaction partners of SakA and MpkC by mass spectrometry analysis. In presence of osmotic stress with sorbitol, 118, and 213 proteins were detected as possible protein interactors of SakA and MpkC, respectively. Under cell wall stress caused by congo red, 420 and 299 proteins were detected interacting with SakA and MpkC, respectively. Interestingly, a group of 78 and 256 proteins were common to both interactome analysis. Co-immunoprecipitation (Co-IP) experiments showed that SakA::GFP is physically associated with MpkC:3xHA upon osmotic and cell wall stresses. We also validated the association between SakA:GFP and the cell wall integrity MAPK MpkA:3xHA and the phosphatase PtcB:3xHA, under cell wall stress. We further characterized A. fumigatus PakA, the homolog of the S. cerevisiae sexual developmental serine/threonine kinase Ste20, as a component of the SakA/MpkC MAPK pathway. The ApakA strain is more sensitive to cell wall damaging agents as congo red, calcofluor white, and caspofungin. Together, our data supporting the hypothesis that SakA and MpkC are part of an osmotic and general signal pathways involved in regulation of the response to the cell wall damage, oxidative stress, drug resistance, and establishment of infection. This manuscript describes an important biological resource to understand SakA and MpkC protein interactions. Further investigation of the biological roles played by these protein interactors will provide more opportunities to understand and combat IA. (AU)

• FAPESP's process: 16/07870-9 - The influence of mitogen activated protein kinases (MAPK) on the expression of genetic determinants important for Aspergillus fumigatus virulence
• Grantee: Gustavo Henrique Goldman
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 17/19288-5 - Characterization of MAPKs dependent-phosphoproteomes of Aspergillus fumigatus under stress conditions
• Grantee: Eliciane Cevolani Mattos
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/00789-6 - Functional characterization of Aspergillus nidulans phosphatases involved on glucose metabolism
• Grantee: Leandro Jose de Assis
• Support Opportunities: Scholarships in Brazil - Post-Doctoral