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Molecular basis of the evolution of alternative tyrosine biosynthetic routes in plants

Nature Chemical Biology volume 13, pages 10291035 (2017) | Download Citation

Abstract

L-Tyrosine (Tyr) is essential for protein synthesis and is a precursor of numerous specialized metabolites crucial for plant and human health. Tyr can be synthesized via two alternative routes by different key regulatory TyrA family enzymes, prephenate dehydrogenase (PDH, also known as TyrAp) or arogenate dehydrogenase (ADH, also known as TyrAa), representing a unique divergence of primary metabolic pathways. The molecular foundation underlying the evolution of these alternative Tyr pathways is currently unknown. Here we characterized recently diverged plant PDH and ADH enzymes, obtained the X-ray crystal structure of soybean PDH, and identified a single amino acid residue that defines TyrA substrate specificity and regulation. Structures of mutated PDHs co-crystallized with Tyr indicate that substitutions of Asn222 confer ADH activity and Tyr sensitivity. Reciprocal mutagenesis of the corresponding residue in divergent plant ADHs further introduced PDH activity and relaxed Tyr sensitivity, highlighting the critical role of this residue in TyrA substrate specificity that underlies the evolution of alternative Tyr biosynthetic pathways in plants.

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Acknowledgements

We thank K. Wallin (University of Minnesota) and U. (Rosia) Schmidt for the cloning of MtncADH and SlncADH, respectively, and P.-M. Delaux (Universite´ de Toulouse) for help with a legume phylogenetic analysis. This work was supported by the National Science Foundation (IOS-1354971 to H.A.M. and MCB-1614539 to J.M.J.). C.K.H. was supported by the NSF Graduate Research Fellowship Program (DGE-1143954). Portions of this research were carried out at the Argonne National Laboratory Structural Biology Center of the Advanced Photon Source, a national user facility operated by the University of Chicago for the Department of Energy Office of Biological and Environmental Research (DE-AC02-06CH11357).

Author information

Author notes

    • Craig A Schenck
    •  & Cynthia K Holland

    These authors contributed equally to this work.

Affiliations

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

    • Craig A Schenck
    • , Matthew R Schneider
    • , Yusen Men
    •  & Hiroshi A Maeda
  2. Department of Biology, Washington University in St. Louis, St. Louis, Missouri, USA.

    • Cynthia K Holland
    • , Soon Goo Lee
    •  & Joseph M Jez

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Contributions

C.A.S., C.K.H., J.M.J., and H.A.M. designed the research, C.A.S. performed phylogenetic analyses, and C.A.S., M.R.S., and Y.M. performed site-directed mutagenesis and biochemical characterization of recombinant enzymes. C.K.H. expressed, purified, and crystallized proteins, collected diffraction data, and determined crystal structures. S.G.L. assisted with SAD phasing, C.A.S., C.K.H., J.M.J., and H.A.M. analyzed data, and C.A.S., C.K.H., J.M.J., and H.A.M. wrote the paper with all authors providing editorial input.

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Hiroshi A Maeda.

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Results, Supplementary Tables 1–3, Supplementary Figures 1–8

Excel files

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    Supplementary Dataset 1

    Sequences used in the phylogenetic analyses from Figure 1 and Supplementary Figure 1

Text files

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    Supplementary Dataset 2

    Full amino acid alignment made in MUSCLE used to construct the phylogenetic analyses in Supplementary Fig. 1

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DOI

https://doi.org/10.1038/nchembio.2414

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