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Emerging function for a glutathione transferase in response to wood extractives toxicity
2014 - IRG/WP 14-10824
The first steps of wood degradation by fungi lead to the release of toxic compounds known as extractives. To better understand how lignolytic fungi cope with these products, a transcriptomic analysis of Phanerochaete chrysosporium genes was performed in presence of oak acetonic extractives. The most up-regulated genes relate to nutrition, nucleic acid modification, gene regulation, signalling and stress responses. Focusing on the induced antioxidant and detoxification systems, a glutathione transferase of the GTT2 class has been selected for functional characterization. This enzyme does not possess any classical glutathione transferase activity but rather a peroxidase activity. The occurrence of the GTT2 gene within the genome is closely linked to the wood decay capabilities of the fungi. This example suggests that the intracellular detoxification system could have evolved concomitantly with the extracellular ligninolytic machinery in relation to the capacity to degrade wood.
A Thuillier, K Chibani, A Deroy, S Dumarçay, P Gérardin, A Kohler, J-P Jacquot, E Gelhaye, M Morel-Rouhier

Interactions between fungal glutathione transferases and wood extractives
2017 - IRG/WP 17-10894
The interactions between extractives from several tree species of French Guyana and glutathione transferases (GST) from the white-rot Trametes versicolor have been studied. These interactions have been monitored using a thermal shift assay. It appears from these experiments that TvGSTO2S interacts strongly with acetonic extract of Bagassa guianensis and in particular with oxyresveratrol. In agreement with previous results, the obtained data suggest that the GST network could give insights on the adaptation of wood decaying fungi to their chemical environment.
T Perrot, F Saiag, N Amusant, S Dumarçay, P Gérardin, M Morel-Rouhier, R SormaniI, E Gelhaye

Interactions between wood polyphenols and detoxification enzymes of the white rot Trametes versicolor
2018 - IRG/WP 18-10906
Wood decay fungi have complex detoxification systems that enable them to cope with secondary metabolites produced by plants. Although the number of genes encoding for glutathione transferases (GSTs) is especially expanded in lignolytic fungi, little is known about their physiological target molecules. In this study, by combining thermal shift assay and affinity crystallography we highlighted interaction between polyphenols and GSTs Omega from the white-rot fungus Trametes versicolor (TvGSTOs). Two distinct ligand-binding sites were found, an open valley at the dimer interface and a deep closed hydrophobic site that corresponds to a peculiar H-site. This site appears to be optimized for aromatic ligand binding, and retains the flavonoid dihydrowogonin from a partially purified wild-cherry extract. Enzymatic inhibition assay and X-ray crystallography confirmed TvGSTOs affinity for polyphenols. The non-catalytic interaction observed suggests that TvGSTOs could be involved in the transport of polyphenols as reported for plant GSTs which have been associated with the transport of anthocyanins into the vacuole.
M Schwartz, T Perrot, E Aubert, S Dumarçay, F Favier, P Gérardin, M Morel-Rouhier, G Mulliert, F Saiag, C Didierjean, E Gelhaye

Glutathione transferases as functional biomarkers of wood degradation?
2010 - IRG/WP 10-10735
The recent release of several fungal genome sequences allows the report of an overview of fungal glutathione transferases (GSTs) focused on their function in the wood-degrading basidiomycete Phanerochaete chrysosporium. A genomic and phylogenetic analysis of GST classes in various sequenced fungi –zygomycetes, ascomycetes and basidiomycetes- revealed some specificity in GSTs distribution, in comparison with previous analysis on ascomycetes. In addition to the classical identified families, this analysis highlighted new GSTs classes of Omega- and Etherase-types. Some of these new GST-types have been produced and purified as recombinant proteins in Escherichia coli and their enzymatic activities have been determined using various substrates. In addition, the expression of their corresponding genes was analyzed in P. chrysosporium grown in conditions of oxidative stress and on wood. Of interest, a member of the etherase family displays a high peroxidase activity against cumene peroxides. Furthermore, while the different genes coding for etherases are highly expressed in P. chrysosporium in liquid cultures, one is strongly induced when the fungus was grown on wood compared to malt agar medium. From these results, it appears that some of these encoding-GSTs genes could be potentially used as functional markers.
E Gelhaye, M Morel

Connecting fungal genomic variations to wood chemical composition: A way to understand the adaptation of wood decaying fungi
2012 - IRG/WP 12-10776
The development of new sequencing methods allows now exploring the detoxication systems of wood-decaying fungi. These systems, called xenome, are mainly composed of oxidative enzymes (phase I) and of conjugation enzymes (phase II). These enzymes belong to superfamilies, which are extended in the plant cell-wall interacting fungi. From different approaches coupling in particular comparative genomic and biochemical and structural protein biochemistry, we develop in this paper the idea that the expanded xenomic network could reflect the adaptation of these wood-decaying fungi to their molecular environment and particularly to the wood chemical composition.
E Gelhaye, Y Mathieu, E Meux, A Thuillier, M Mélanie

Glutathione transferases and wood extracts
2015 - IRG/WP 15-10845
Wood rotting fungi have developed an efficient detoxification system allowing them to be well adapted to their way of life, these fungi generating and encountering potential toxic compounds during wood degradation. Belonging to this detoxification network, glutathione transferases are particularly extended in these fungi and their presence could reflect the chemical environments encountered by the fungi. To test this hypothesis, we developed biochemical assays to quantify interactions between wood extracts and glutathione transferases from from two white-rot fungi: Trametes versicolor and Phanerochaete chrysosporium. The obtained preliminary results reported in this paper demonstrate that GSTs from white-rot fungi could be used to discriminate wood extracts from different origins in accordance with their chemical compositions.
A Deroy, F Saiag, Z Kebbi-Benkeder, N Touahri, M Morel-Rouhier, F Colin, S Dumarcay, P Gérardin,

From fungal detoxification systems to wood durability in neotropical forests
2019 - IRG/WP 19-10946
White-rot fungi are able to mineralize all the wood components due in particular to an efficient detoxification system. We hypothesised that components of this detoxification systems, glutathione transferases, could be used as tools to explore the natural durability of neotropical wood species. Analysis of the interactions between six glutathione transferases of Trametes versicolor and extracts of 21 wood species from French Guiana revealed a positive correlation between natural durability of the tested wood species (soil tests) and these interactions (Glutathione Transferase Assay). The obtained data suggest that the developed biochemical test could be used to estimate wood natural durability
T Perrot, G Salzet, N Amusant, J Duchene, E Gelhaye