Article

Non-plastidic, tyrosine-insensitive prephenate dehydrogenases from legumes

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Abstract

L-Tyrosine (Tyr) and its plant-derived natural products are essential in both plants and humans. In plants, Tyr is generally assumed to be synthesized in the plastids via arogenate dehydrogenase (TyrAa, also known also ADH), which is strictly inhibited by L-Tyr. Using phylogenetic and expression analyses, together with recombinant enzyme and endogenous activity assays, we identified prephenate dehydrogenases (TyrAps, also known as PDHs) from two legumes, Glycine max (soybean) and Medicago truncatula. The identified PDHs were phylogenetically distinct from canonical plant ADH enzymes, preferred prephenate to arogenate substrate, localized outside of the plastids and were not inhibited by L-Tyr. The results provide molecular evidence for the diversification of primary metabolic Tyr pathway via an alternative cytosolic PDH pathway in plants.

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Acknowledgements

We thank R. Singhal (University of Wisconsin) for help with protoplast isolation; S. Swanson, A. Chanoca and R. Buono (University of Wisconsin) for assistance with confocal imaging and analysis; C. Grau (University of Wisconsin) for soybean Williams82 seeds; J.-M. Ané (University of Wisconsin) for M. truncatula tissues and seeds; and F.R. Márquez (Universidad Andrés Bello) for the 35S-pPZP211 vector. The confocal imaging was performed at the Newcomb Imaging Center, Department of Botany, University of Wisconsin-Madison. This work was supported by grant IOS-1354971 from the US National Science Foundation to H.A.M., the Fritz Went Undergraduate Fellowship to S.C. and the start-up funds from the Graduate School, the College of Letters & Science, and the Department of Botany, University of Wisconsin-Madison to H.A.M.

Author information

Affiliations

  1. Department of Botany, University of Wisconsin–Madison, Madison, Wisconsin, USA.

    • Craig A Schenck
    • , Siyu Chen
    •  & Hiroshi A Maeda
  2. DuPont Pioneer, Hayward, California, USA.

    • Daniel L Siehl

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Contributions

C.A.S. and H.A.M. designed research, and C.A.S. performed phylogenetic analyses, enzyme purification and characterization, localization studies and inhibition assays. S.C. performed gene expression analyses, enzyme purification and enzymatic assays. D.L.S. performed chromatographic separations and enzymatic assays, and H.A.M. performed localization studies and enzymatic assays. C.A.S., S.C., D.L.S. and H.A.M. analyzed data, C.A.S. and H.A.M. wrote the manuscript. All authors read and edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hiroshi A Maeda.

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    Supplementary Text and Figures

    Supplementary Results, Supplementary Tables 1–3 and Supplementary Figures 1–13.