Type

Journal Article

Authors

Sean Doyle
Martin Clynes
Eoin Mulvihill
Stephen Dolan
Michael Henry
Paula Meleady
Rebecca A Owens
Nicola M Moloney

Subjects

Microbiology

Topics
membrane proteins iron acquisition proteome humans iron environment biosynthesis aspergillus fumigatus

The iron-responsive microsomal proteome of Aspergillus fumigatus. (2015)

Abstract Aspergillus fumigatus is an opportunistic fungal pathogen. Siderophore biosynthesis and iron acquisition are essential for virulence. Yet, limited data exist with respect to the adaptive nature of the fungal microsomal proteome under iron-limiting growth conditions, as encountered during host infection. Here, we demonstrate that under siderophore biosynthetic conditions - significantly elevated fusarinine C (FSC) and triacetylfusarinine C (TAFC) production (p<0.0001), extensive microsomal proteome remodelling occurs. Specifically, a four-fold enrichment of transmembrane-containing proteins was observed with respect to whole cell lysates following ultracentrifugation-based microsomal extraction. Comparative label-free proteomic analysis of microsomal extracts isolated following iron-replete and -deplete growth identified 710 unique proteins. Scatterplot analysis (MaxQuant) demonstrated high correlation amongst biological replicates from each growth condition (Pearson correlation >0.96 within groups; biological replicates (n=4). Quantitative and qualitative comparison revealed 231 proteins with a significant change in abundance between the iron-replete and iron-deplete conditions (p<0.05, fold change ≥2), with 96 proteins showing increased abundance and 135 with decreased abundance following iron limitation, including predicted siderophore transporters. Fluorescently labelled FSC was only sequestered following A. fumigatus growth under iron-limiting conditions. Interestingly, human sera exhibited significantly increased reactivity (p<0.0001) against microsomal protein extracts obtained following iron-deplete growth. The opportunistic fungal pathogen Aspergillus fumigatus must acquire iron to facilitate growth and pathogenicity. Iron-chelating non-ribosomal peptides, termed siderophores, mediate iron uptake via membrane-localised transporter proteins. Here we demonstrate for the first time that growth of A. fumigatus under iron-deplete conditions, concomitant with siderophore biosynthesis, leads to an extensive remodelling of the microsomal proteome which includes significantly altered levels of 231 constituent proteins (96 increased and 135 decreased in abundance), many of which have not previously been localised to the microsome. We also demonstrate the first synthesis of fluorescent version of fusarinine C, an extracellular A. fumigatus siderophore, and its uptake and localization under iron-restricted conditions. This infers the use of an A. fumigatus siderophore as a 'Trojan horse' to potentiate the efficacy of anti-fungal drugs. Finally, in addition to revealing the Aspergillus-specific IgG reactivity in normal human sera against microsomal proteins, there appears to be a significantly increased reactivity against microsomal proteins obtained following iron-restricted growth. We hypothesise that iron-limiting environment in humans, which has evolved to nutritionally limit pathogen growth in vivo, may also alter the fungal microsomal proteome.
Collections Ireland -> Dublin City University -> PubMed

Full list of authors on original publication

Sean Doyle, Martin Clynes, Eoin Mulvihill, Stephen Dolan, Michael Henry, Paula Meleady, Rebecca A Owens, Nicola M Moloney

Experts in our system

1
Sean Doyle
Maynooth University
Total Publications: 119
 
2
Martin Clynes
Dublin City University
Total Publications: 209
 
3
Eoin Mulvihill
Maynooth University
Total Publications: 4
 
4
Stephen K Dolan
Maynooth University
Total Publications: 21
 
5
Michael Henry
Dublin City University
Total Publications: 78
 
6
Paula Meleady
Dublin City University
Total Publications: 95
 
7
Rebecca A Owens
Maynooth University
Total Publications: 22
 
8
Nicola M Moloney
Maynooth University
Total Publications: 5