Aspergillus fumigatushigh osmolarity glycerol mitogen activated protein kinases saka and mpkc physically interact during osmotic and cell wall stresses

Adriana Oliveira Manfiolli, Eliciane Cevolani Mattos, Leandro José De Assis, Lilian Pereira Silva, Mevlüt Ulas, Neil Andrew Brown, Rafael Silva-Rocha, Özgür Bayram, Gustavo H. Goldman

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 δpakA 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.

Original languageEnglish
Article number918
Pages (from-to)1-17
Number of pages17
JournalFrontiers in Microbiology
Volume10
DOIs
Publication statusPublished - 7 May 2019

Keywords

  • Aspergillus fumigatus
  • HOG
  • Mitogen activate protein kinase
  • MPKC
  • SakA

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Aspergillus fumigatushigh osmolarity glycerol mitogen activated protein kinases saka and mpkc physically interact during osmotic and cell wall stresses. / Manfiolli, Adriana Oliveira; Mattos, Eliciane Cevolani; De Assis, Leandro José; Silva, Lilian Pereira; Ulas, Mevlüt; Brown, Neil Andrew; Silva-Rocha, Rafael; Bayram, Özgür; Goldman, Gustavo H.

In: Frontiers in Microbiology, Vol. 10, 918, 07.05.2019, p. 1-17.

Research output: Contribution to journalArticle

Manfiolli, Adriana Oliveira ; Mattos, Eliciane Cevolani ; De Assis, Leandro José ; Silva, Lilian Pereira ; Ulas, Mevlüt ; Brown, Neil Andrew ; Silva-Rocha, Rafael ; Bayram, Özgür ; Goldman, Gustavo H. / Aspergillus fumigatushigh osmolarity glycerol mitogen activated protein kinases saka and mpkc physically interact during osmotic and cell wall stresses. In: Frontiers in Microbiology. 2019 ; Vol. 10. pp. 1-17.
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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 δpakA 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.",
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AU - Manfiolli, Adriana Oliveira

AU - Mattos, Eliciane Cevolani

AU - De Assis, Leandro José

AU - Silva, Lilian Pereira

AU - Ulas, Mevlüt

AU - Brown, Neil Andrew

AU - Silva-Rocha, Rafael

AU - Bayram, Özgür

AU - Goldman, Gustavo H.

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AB - 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 δpakA 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.

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