Aspergillus fumigatus is an opportunistic and allergenic pathogenic fungus, responsible for fungal infections in humans. A. fumigatus 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 integrity of the cell wall and leading to fungal death. In some A. fumigatus strains, caspofungin treatment at high concentrations induces an increase of fungal growth, a phenomenon called the caspofungin paradoxical effect (CPE). Here, we analyze the proteome and phosphoproteome of the A. fumigatus wild-type strain and of mitogen-activated protein kinase (MAPK) mpkA and sakA null mutant strains during CPE (2 μg/ml caspofungin for 1 h). The wild-type proteome showed 75 proteins and 814 phosphopeptides (corresponding to 520 proteins) altered in abundance in response to caspofungin treatment. The ΔmpkA (ΔmpkA caspofungin/wild-type caspofungin) and ΔsakA (ΔsakA caspofungin/wild-type caspofungin) strains displayed 626 proteins and 1,236 phosphopeptides (corresponding to 703 proteins) and 101 proteins and 1,217 phosphopeptides (corresponding to 645 proteins), respectively, altered in abundance. Functional characterization of the phosphopeptides from the wild-type strain exposed to caspofungin showed enrichment for transcription factors, protein kinases, and cytoskeleton proteins. Proteomic analysis of the ΔmpkA and ΔsakA mutants indicated that control of proteins involved in metabolism, such as in production of secondary metabolites, was highly represented in both mutants. Results of functional categorization of phosphopeptides from both mutants were very similar and showed a high number of proteins with decreased phosphorylation of proteins involved in transcriptional control, DNA/RNA binding, cell cycle control, and DNA processing. This report reveals novel transcription factors involved in caspofungin tolerance.

中文翻译：

---

暴露于抗真菌药卡泊芬净的烟曲霉的蛋白质组学研究。

烟曲霉是一种机会性和致敏性致病真菌，负责人类的真菌感染。烟曲霉感染通常用多烯，唑类或棘孢菌素治疗。Echinocandins，例如卡泊芬净，可以抑制β-1,3-葡聚糖多糖的生物合成，影响细胞壁的完整性并导致真菌死亡。在一些烟曲霉菌株，在高浓度卡泊芬净处理诱导真菌生长的增加，这种现象叫做Ç aspofungin p aradoxical ë ffect（CPE）。在这里，我们分析了烟曲霉的蛋白质组和磷酸化蛋白质组。野生型菌株以及CPE期间的促分裂原活化蛋白激酶（MAPK）mpkA和sakA无效突变菌株（2μg/ ml卡泊芬净1小时）。野生型蛋白质组显示了75种蛋白质和814种磷酸肽（相当于520种蛋白质）响应卡泊芬净治疗而发生了丰度变化。该Δ mpkA（Δ mpkA卡泊芬净/野生型卡泊芬净）和Δ坂（Δ坂卡泊芬净/野生型卡泊芬净）菌株分别显示了626个蛋白和1,236个磷酸肽（对应703个蛋白）和101个蛋白和1,217个磷酸肽（对应645个蛋白），其丰度发生了变化。来自暴露于卡泊芬净的野生型菌株的磷酸肽的功能表征显示出转录因子，蛋白激酶和细胞骨架蛋白的富集。ΔmpkA和ΔsakA的蛋白质组学分析突变体表明，在两个突变体中都高度控制了参与代谢的蛋白质，例如次级代谢产物的产生。来自两个突变体的磷酸肽的功能分类结果非常相似，并且显示出大量蛋白质，其中涉及转录控制，DNA / RNA结合，细胞周期控制和DNA加工的蛋白质的磷酸化降低。该报告揭示了卡泊芬净耐受性中涉及的新型转录因子。