Phosphoproteomics of Aspergillus fumigatus exposed to the antifungal drug caspofungin, Supplementary files

<em>Aspergillus fumigatus</em> is an opportunistic and allergenic pathogenic fungus, responsible for fungal infections in humans. <em>A. fumigatus </em>infections are usually treated with polyenes, azoles, or echinocandins. Echinocandins, such as caspofungin, can inhibit the biosynthesis of the β-1,3-glucan polysaccharide, affecting the cell wall integrity and leading to fungal death. In some <em>A. fumigatus </em>strains, caspofungin treatment at high concentrations induces an increase of fungal growth, a phenomenon called Caspofungin Paradoxical Effect (CPE). Here, we analyze the proteome and phosphoproteome of <em>A. fumigatus</em> wild-type and the mitogen-activated protein kinases <em>mpkA</em> and <em>sakA </em>null mutant strains during CPE (2 µg/ml caspofungin for 1 hour). The wild-type proteome shows 75 proteins and 814 phosphopeptides (corresponding to 520 proteins) altered in abundance in response to caspofungin treatment. The <em>ΔmpkA </em>(<em>ΔmpkA </em>caspofungin/wild-type caspofungin) and <em>ΔsakA</em> (<em>ΔsakA </em>caspofungin/wild-type caspofungin) display 626 proteins and 1236 phosphopeptides (corresponding to 703 proteins) and 101 proteins and 1217 phosphopeptides (corresponding to 645 proteins), respectively, altered in abundance. Functional characterization of the phosphopeptides from the wild-type exposed to caspofungin show enrichment for transcription factors, protein kinases, and cytoskeleton proteins. Proteomic analysis for the Δ<em>mpkA</em> and Δ<em>sakA</em> mutants indicates that the control of proteins involved in metabolism, such as in secondary metabolites production, was highly represented in both mutants. Functional categorization of phosphopeptides from both mutants is very similar, and show a high number of proteins with decreased phosphorylation of proteins involved in transcriptional control, DNA/RNA binding, cell cycle control, and DNA processing. This study reveals novel transcription factors involved in caspofungin tolerance.