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Translational Regulation of Metabolic Dynamics during Effector-Triggered Immunity

ABSTRACT: It is not fully known whether translational regulation also occurs in later stage immune responses, such as effector-triggered immunity (ETI), which often leads to strong metabolic dynamics. In this study, we performed a genome-wide ribosome profiling in Arabidopsis upon ETI activation and discovered that specific groups of genes were translationally regulated, especially metabolic genes in aromatic amino acid, phenylpropanoid, camalexin, and sphingolipid metabolism. The involvement of these components in the induction of ETI was confirmed by genetic analysis, amino acid profiling and exogeneous application of phenylalanine or an inhibitor of aromatic amino acid biosynthesis. Our findings provide new insight into the diverse translational regulation in the plant immune responses and demonstrate that translational coordination of metabolic gene expression is an important strategy for ETI activation.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE124115 | GEO | 2020/12/16

REPOSITORIES: GEO

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Project description:Its characteristic rose-like aroma makes phenylethanol a popular ingredient in foods, beverages and cosmetics. Microbial production of phenylethanol currently relies on whole-cell bioconversion of phenylalanine with yeasts that harbor an Ehrlich pathway for phenylalanine catabolism. Complete biosynthesis of phenylethanol from a cheap carbon source such as glucose provides an economically attractive alternative for phenylalanine bioconversion. In this study, a Synthetic Genetic Array screening was applied to identify genes involved in regulation of phenylethanol synthesis in Saccharomyces cerevisiae. The screen focused on transcriptional regulation of ARO10, which encodes the major decarboxylase involved in conversion of phenylpyruvate to phenylethanol. A deletion in ARO8, which encodes an aromatic amino acid transaminase, was found to cause a transcriptional upregulation of ARO10 during growth with ammonium sulfate as the sole nitrogen source. Physiological characterization revealed that the aro8M-oM-^AM-^D mutation led to substantial changes in the absolute and relative intracellular concentrations of amino acids. Moreover, deletion of ARO8 led to de novo production of phenylethanol during growth on a glucose synthetic medium with ammonium as the sole nitrogen source. The aro8 mutation also stimulated phenylethanol production when combined with other, previously documented mutations that deregulate aromatic amino acid biosynthesis in S. cerevisiae. The resulting engineered S. cerevisiae strain produced over 3 mM of phenylethanol from glucose during growth on a simple synthetic medium. The strong impact of a transaminase deletion on intracellular amino acid concentrations opens new possibilities for yeast-based production of amino acid-derived products. The goal of the present study was to identify genes that influence the transcriptional (de)repression of the Ehrlich pathway during growth with ammonium as the nitrogen source. With the aid of Synthetic Genetic Array technology, we constructed a strain collection in which deletions in the non-essential genes in the S. cerevisiae genome were combined with a reporter plasmid comprising the ARO10 promoter fused to a reporter gene (egfp) encoding a fluorescent reporter protein. After screening by flow cytometry, deletion of ARO8 led to a deregulated expression from the ARO10 promoter. The impact of this deletion was further studied by transcriptome and intracellular metabolite analyses. Furthermore, phenylethanol production was measured in strains that combined the aro8 mutation with mutations that were previously shown to deregulate aromatic amino acid biosynthesis.

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Translational Regulation of Metabolic Dynamics during Effector-Triggered Immunity

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