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Designed abscisic acid analogs as antagonists of PYL-PP2C receptor interactions

Nature Chemical Biology volume 10, pages 477–482 (2014)Cite this article

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Abstract

The plant stress hormone abscisic acid (ABA) is critical for several abiotic stress responses. ABA signaling is normally repressed by group-A protein phosphatases 2C (PP2Cs), but stress-induced ABA binds Arabidopsis PYR/PYL/RCAR (PYL) receptors, which then bind and inhibit PP2Cs. X-ray structures of several receptor–ABA complexes revealed a tunnel above ABA's 3′ ring CH that opens at the PP2C binding interface. Here, ABA analogs with sufficiently long 3′ alkyl chains were predicted to traverse this tunnel and block PYL-PP2C interactions. To test this, a series of 3′-alkylsulfanyl ABAs were synthesized with different alkyl chain lengths. Physiological, biochemical and structural analyses revealed that a six-carbon alkyl substitution produced a potent ABA antagonist that was sufficiently active to block multiple stress-induced ABA responses in vivo. This study provides a new approach for the design of ABA analogs, and the results validated structure-based design for this target class.

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Figure 1: Synthesis of ASn compounds and ASn effects on Arabidopsis seed germination.
Figure 2: Effects of ASn compounds on expression of Arabidopsis ABA-responsive genes.
Figure 3: Effects of ASn on HAB1 inhibition by ABA receptors.
Figure 4: Crystal structure of PYR1–AS6 complex.

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Acknowledgements

We thank T. Nakagawa for providing pGWB vectors and Toray Industries Inc., Tokyo, Japan, for the gift of (+)-ABA. This work was supported in part by the Japanese Society for the Promotion of Science (JSPS) Postdoctoral Fellowships for Research Abroad (to M.O.), JSPS KAKENHI (Grant-in-Aid for Young Scientists 26711018 to M.O.) and the US National Science Foundation (Integrative Organismal Systems 0820508 to S.R.C.).

Author information

Author notes

  1. Akira Endo

    Present address: Present address: Crop Breeding Research Division, National Agricultural Research Organization for Hokkaido Region, Hokkaido, Japan.,

  2. Jun Takeuchi and Masanori Okamoto: These authors contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Science and Technology, Shizuoka University, Shizuoka, Japan

    Jun Takeuchi & Yasushi Todoroki

  2. Arid Land Research Center, Tottori University, Tottori, Japan

    Masanori Okamoto

  3. Department of Botany and Plant Sciences and Center for Plant Cell Biology, University of California–Riverside, Riverside, California, USA

    Masanori Okamoto & Sean R Cutler

  4. Department of Bioscience, Tokyo University of Agriculture, Tokyo, Japan

    Tomonori Akiyama & Shunsuke Yajima

  5. Graduate School of Agriculture, Shizuoka University, Shizuoka, Japan

    Takuya Muto, Toshiyuki Ohnishi & Yasushi Todoroki

  6. Department of Applied Biology and Chemistry, Tokyo University of Agriculture, Tokyo, Japan

    Masayuki Sue

  7. RIKEN Center for Sustainable Resource Science, Kanagawa, Japan

    Mitsunori Seo & Yuri Kanno

  8. National Institute for Agro-Environmental Sciences, Ibaraki, Japan

    Tsunashi Kamo

  9. Department of Cell and Systems Biology, University of Toronto, Toronto, Ontario, Canada

    Akira Endo & Eiji Nambara

  10. Graduate School of Agriculture, Kyoto University, Kyoto, Japan

    Nobuhiro Hirai

  11. Research Institute of Green Science and Technology, Shizuoka University, Shizuoka, Japan

    Toshiyuki Ohnishi & Yasushi Todoroki

Authors
  1. Jun Takeuchi
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Contributions

J.T., M.O. and Y.T. conceived and J.T., M.O., S.R.C. and Y.T. designed the research. J.T., M.O., T.A., T.M., S.Y., M. Sue, M. Seo, Y.K., K.T., A.E., E.N., S.R.C. and Y.T. performed the research and/or analyzed data. J.T., M.O., S.R.C. and Y.T. co-wrote the manuscript with the assistance of M. Sue, M. Seo, N.H. and T.O.

Corresponding author

Correspondence to Yasushi Todoroki.

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The authors declare no competing financial interests.

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Takeuchi, J., Okamoto, M., Akiyama, T. et al. Designed abscisic acid analogs as antagonists of PYL-PP2C receptor interactions. Nat Chem Biol 10, 477–482 (2014). https://doi.org/10.1038/nchembio.1524

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