Leucine metabolism regulates TRI6 expression and affects deoxynivalenol production and virulence in Fusarium graminearum
Subramaniam, R., Narayanan, S., Walkowiak, S., Wang, L., Joshi, M., Rocheleau, H., Ouellet, T., Harris, L.J. (2015). Leucine metabolism regulates TRI6 expression and affects deoxynivalenol production and virulence in Fusarium graminearum, 98(4), 760-769. http://dx.doi.org/10.1111/mmi.13155
© 2015 John Wiley & Sons Ltd. TRI6 is a positive regulator of the trichothecene gene cluster and the production of trichothecene mycotoxins [deoxynivalenol (DON)] and acetylated forms such as 15-Acetyl-DON) in the cereal pathogen Fusarium graminearum. As a global transcriptional regulator, TRI6 expression is modulated by nitrogen-limiting conditions, sources of nitrogen and carbon, pH and light. However, the mechanism by which these diverse environmental factors affect TRI6 expression remains underexplored. In our effort to understand how nutrients affect TRI6 regulation, comparative digital expression profiling was performed with a wild-type F.graminearum and a Δtri6 mutant strain, grown in nutrient-rich conditions. Analysis showed that TRI6 negatively regulates genes of the branched-chain amino acid (BCAA) metabolic pathway. Feeding studies with deletion mutants of MCC, encoding methylcrotonyl-CoA-carboxylase, one of the key enzymes of leucine metabolism, showed that addition of leucine specifically down-regulated TRI6 expression and reduced 15-ADON accumulation. Constitutive expression of TRI6 in the Δmcc mutant strain restored 15-ADON production. A combination of cellophane breach assays and pathogenicity experiments on wheat demonstrated that disrupting the leucine metabolic pathway significantly reduced disease. These findings suggest a complex interaction between one of the primary metabolic pathways with a global regulator of mycotoxin biosynthesis and virulence in F.graminearum. Global regulator TRI6 in F. graminearum links primary and secondary metabolism. In nutrient-rich conditions, TRI6, in addition to auto-regulate its expression, also acts as a negative regulator and represses genes of the leucine metabolic pathway. In nutrient-deprived conditions, TRI6 morphs into a positive regulator and activate genes involved in trichothecene biosynthesis.
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