Bacterial-fungal confrontations after 66 hours of incubation.

<p>Different letters above boxplots indicate significant difference based on ANOVA followed by Tukey's HSD test (<i>p</i><0.05). <b>A.</b> Ratio R (ratio of the diameter of the fungal colony growing in the direction of the bacterial colonies over the diameter of the colony growing in the orthogonal direction, see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0147100#sec002" target="_blank">Materials and Methods</a>) of the fungal growth in presence of one bacterial strain, as a function of the treatment. <i>n</i> = 25 bacterial strains per treatment with 5 replicates per strain. Control corresponded to <i>P</i>. <i>chrysosporium</i> growing alone. <b>B.</b> Relationships between the genus of the bacterial strains and the effect on the fungal growth based on the R ratio. For <i>Burkholderia</i>, <i>n</i> = 90 strains; for <i>Collimonas</i>, <i>n</i> = 3; for <i>Dyella</i>, <i>n</i> = 8; for <i>Luteibacter</i>, <i>n</i> = 23. <i>Cupriavidus</i> was excluded from the analysis because <i>n</i> = 1.</p

Metabolic profiles of 125 bacterial strains randomly selected in the 5 treatments (E0, E1B, E1BF, E3B, E3BF) (<i>n</i> = 25).

<p>The heatmap shows the 51/95 substrates with statistically significant differences between treatments (enrichment step E x presence (BF) or absence (B) of <i>P</i>. <i>chrysosporium</i>) (ANOVA, <i>p</i><0.05).</p

16S rRNA gene-based dendrogram generated from the UPGMA clustering based on unweighted UniFrac distances.

<p>Branches are proportional to this distance. <i>n</i> = 25 sequences per treatment.</p

Capabilities of the strains able to grow on the selective media to use different substrates.

<p>Different letters above barplots indicate significant difference based on glm under a binomial distribution (<i>p</i><0.05). <b>A.</b> Cellulolytic activity on carboxymethyl-cellulose medium. <i>n</i> = 25 bacterial strains per treatment with 3 replicates per strain. <b>B.</b> Xylanolytic activity on beechwood xylan medium. <i>n</i> = 25 bacterial strains per treatment with 3 replicates per strain. <b>C.</b> Siderophore production assay on CAS medium. <i>n</i> = 22 bacterial strains for E0, <i>n</i> = 10 for E1B, <i>n</i> = 18 for E1BF, <i>n</i> = 19 for E3B, <i>n</i> = 22 for E3BF; with 3 replicates for each strain. Variation in the number of strains is due to the fact that not all strains could grow on CAS medium.</p

supp data sequences

fasta file of the sequences used for the phylogenetic analysi

ScECM4 glutathione binding site.

<p>The catalytic cysteine C46, the residues involved in polar interactions with glutathione (GS) and the tyrosines Y224, Y301 and Y346 from the triad are represented as sticks. Polar interactions (yellow dashed lines) were observed between: E173 carboxylate group and GSH N-terminal amine group; NH of S174 and carboxylate group of GSH γ-glutamyl residue; S174 hydroxyl group and carboxylate group of GSH γ-glutamyl residue; R15 guanidinium group and carbonyl group of GSH γ-glutamyl residue; W79 indole group and carbonyl group of GSH cysteine; NHs and COs of V158 and of GSH cysteine; R155 guanidinium group and carboxylate group of GSH glycinyl residue.</p

ScECM4 monomer with its specific features.

<p>The monomer of ScECM4 is represented as cartoon (N-terminal domain in cyan, C-terminal domain in violet). The N-terminal end is colored in wheat, the loop β2-α2 is colored in red and the extension of helix α5 along with helix α5’ are colored in blue.</p

Structural alignment of the GSTXs available in the PDB and corresponding secondary structure elements.

<p>Structural alignment of 5 GSTXs available in the Protein Data Bank (PDB codes PcGHR1 3PPU, PtGHR1 4USS, EcYqjG 4G0K, ScPcpF 4FQU, GbGSTX 4PTS) with highlight of the secondary structure elements (helices in yellow, sheets in gray). Catalytic residues (the conserved catalytic cysteine and the triad of tyrosines) are highlighted in black. The symbols <b>$</b> indicate the positions of the residues that form interactions with the glutathione and the symbols <b>#</b> indicate the positions of the putative (hydro)quinone binding site residues. PtGHR1: GHR from <i>Populus trichocarpa</i>, PcGHR1: GHR1 from <i>Phanerochaete chrysosporium</i>, EcYqjG: YqjG from <i>Escherichia coli</i>, ScPcpF: PcpF from <i>Sphingobium chlorophenolicum</i>, GbGSTX: putative GHR from <i>Gordonia bronchialis</i>.</p

Activity of ScECM4 with GS-menadione.

<p><b>Top,</b> UV/visible spectrum of 100 μM GS-menadione in Tris-EDTA buffer, as recorded before addition of 1 μM ScECM4 and 1 mM GSH (solid line), and then after 10 min of reaction with these compounds (dotted line). <b>Bottom,</b> HPLC profile at 250 nm after 15 min incubation of ScECM4 with GS-menadione and GSH. GS-menadione shows a typical peak at a retention time of 10 min (solid line) and menadione at 12 min (dotted line).</p

ScECM4 active site with superimposed GS-menadione.

<p>Residues of ScECM4 that form putative interactions with GS-menadione are labelled and represented as sticks. In addition to the G-site residues described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0164678#pone.0164678.g005" target="_blank">Fig 5</a>, the drawing highlights the putative roles of W48, F228, H345, H350 and a water molecule (H₂O) for the binding of the (hydro)quinone moiety, depending on its orientation.</p