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Trees were based on phylogeny reconstruction and generated from Muscle Alignments (75) using MEGA 6 software (76) at the following settings: Gap Open, -2.9 Gap Extend, 0 Hydrophobicity Multiplier, 1.2 for the alignment. Neighbor joining trees for Crp1 (A) and Prx1 (B) homologous proteins within the Saccharomycotina. Data represent the mean and standard deviation of three biological replicates with asterisks indicating statistical significance in Student's t-test comparing all conditions to the one w/o DO for the individual strains ( ** p < 0.01). parapsilosis strains was measured as an optical density at a wavelength of 600 nm in minimal medium supplemented with (w) or without (w/o) an amino acid dropout solution (DO) containing (+) or lacking (−) methionine. parapsilosis Δ/Δprx1 is not restored by D-methionine. Error bars indicate the standard deviation. All data show average expression levels relative to time point 0 based on three biological and three technical replicates. parapsilosis was analyzed by qRT-PCR using total RNA isolated after exposure to human monocyte-derived macrophages. Gene expression of PRX1, CRP1 and CTR1 during confrontation with primary human macrophages. * growth of unicellular organism as a thread to attached cells (BP:GO:0070783)

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A full list of genes and enriched GO terms in provided in Suppl. The numbers indicate the number of DEG assign to the particular cluster. Enriched GO terms, displayed in the pie charts, we determined by the Benjamini–Hochberg corrected p-value. Positively and negatively regulated genes we assign to categories according the molecular function (MF), cellular component (CC) and biological process (BP) using an on-line tool Slim Mapper at Candida Genome Database. Cluster analysis of differentially expressed genes in C.

Single images were captured after 30 min of coincubation (A) or were selected from time-lapse microscopy showing the FITC and Lysotracker channels, a brightfield (BF) image and over lays of selected channels (B). aurantium in predator-prey ratio 1:10 and seeded into 4-well Glass Bottom μ-Slides. parapsilosis (fluorescein isothiocyanate, FITC) resides in acidic phagolysomes (Lysotracker) of P. All of them were found to contain signal peptides and these could be the interacting partners with the specific vesicle proteins from P. aurantium with its predicted functional annotation with reference to UniProt. parapsilosis from the vesicles proteome of P. Out of 56 identified proteins, 25 proteins (44.6%) were found to contain signal peptides.ĭataset S3b: Identified proteins of C. Identified proteins from the vesicles proteome of P. Gene ontology annotations for enrichment of defined categories in molecular function, cell component, and biological process.Ĭmi1338-Supinfo3.xls圎xcel 2007 spreadsheetĭataset S3a. aurantium.Ĭmi1338-Supinfo2.xls圎xcel 2007 spreadsheetĭataset S2. parapsilosis during confrontation with P.

Tables of all differentially expressed genes in C. Take AwayĬmi1338-Supinfo1.xls圎xcel 2007 spreadsheetĭataset S1. Although completely unknown proteins were dominant, many of them could be categorised as hydrolytic enzymes targeting the fungal cell wall, indicating that fungal cell wall structures are under selection pressure by predatory phagocytes in natural environments. Proteomic analysis identified 56 vesicular proteins from P. Following their isolation, the content of these vesicles induced immediate killing and lysis of C. The fungicidal activity was largely retained in intracellular vesicles of the amoebae. On the fungal side, a functional genomic approach identified copper and redox homeostasis as primary targets of amoeba predation, with the highly expressed copper exporter gene CRP1 and the peroxiredoxin gene PRX1 contributing to survival when encountered with P. Here we show that upon phagocytic ingestion by the amoeba, Candida parapsilosis is confronted with an oxidative burst and undergoes lysis within minutes of processing in acidified phagolysosomes. The fungivorous amoeba Protostelium aurantium has a wide fungal food spectrum including foremost pathogenic members of the genus Candida. Predatory interactions among microbes are major evolutionary driving forces for biodiversity.

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