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PP2A activates brassinosteroid-responsive gene expression and plant growth by dephosphorylating BZR1

Nature Cell Biology volume 13, pages 124–131 (2011)Cite this article

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Abstract

When brassinosteroid levels are low, the GSK3-like kinase BIN2 phosphorylates and inactivates the BZR1 transcription factor to inhibit growth in plants. Brassinosteroid promotes growth by inducing dephosphorylation of BZR1, but the phosphatase that dephosphorylates BZR1 has remained unknown. Here, using tandem affinity purification, we identified protein phosphatase 2A (PP2A) as a BZR1-interacting protein. Genetic analyses demonstrated a positive role for PP2A in brassinosteroid signalling and BZR1 dephosphorylation. Members of the B’ regulatory subunits of PP2A directly interact with BZR1’s putative PEST domain containing the site of the bzr1-1D mutation. Interaction with and dephosphorylation by PP2A are enhanced by the bzr1-1D mutation, reduced by two intragenic bzr1-1D suppressor mutations, and abolished by deletion of the PEST domain. This study reveals a crucial function for PP2A in dephosphorylating and activating BZR1 and completes the set of core components of the brassinosteroid-signalling cascade from cell surface receptor kinase to gene regulation in the nucleus.

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Figure 1: Identification of PP2A as a BZR1-interacting phosphatase.
Figure 2: Overexpression of PP2AB′α and PP2AB′β activates brassinosteroid signalling and partially suppresses brassinosteroid signalling mutants.
Figure 3: PP2A is essential for brassinosteroid signalling and BZR1 dephosphorylation.
Figure 4: PP2A B′ subunit binds to the PEST domain of BZR1 to promote BZR1 dephosphorylation in vivo.
Figure 5: PP2A dephosphorylates BZR1 and abolishes its binding by the 14-3-3 proteins.

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Acknowledgements

This work was financially supported by the Division of Chemical Sciences, Geosciences, and Biosciences, Office of Basic Energy Sciences of the US Department of Energy through Grant DE-FG02-08ER15973, and by NIH (R01GM066258), National Science Foundation of China (30870213, 90917008), NSF (IOS-0724688 and IOS-0846282), and the Herman Frasch Foundation. R.W and M.Y. were supported by the China Scholarship Council. The UCSF Mass Spectrometry Facility (A.L.B., Director) is supported by the Biomedical Research Technology Program of the National Centre for Research Resources, NIH NCRR RR01614, RR012961 and RR019934.

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  1. Wenqiang Tang, Min Yuan, Ruiju Wang and Yihong Yang: These authors contributed equally to this work

Authors and Affiliations

  1. Institute of Molecular Cell Biology, College of Life Science, Hebei Normal University, Shijiazhuang, Hebei, 050016, China

    Wenqiang Tang, Min Yuan, Yihong Yang, Chunming Wang & Ying Sun

  2. Department of Plant Biology, Carnegie Institution of Washington, Stanford, California 94305, USA

    Wenqiang Tang, Min Yuan, Ruiju Wang, Tae-Wuk Kim, Zhiping Deng, Srinivas S. Gampala, Joshua M. Gendron, Ying Sun & Zhi-Yong Wang

  3. Institute of Molecular Biology, College of Life Science, Nankai University, Tianjin 300071, China

    Ruiju Wang

  4. Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94143, USA

    Juan A. Oses-Prieto & Alma L. Burlingame

  5. Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island 02912, USA

    Hong-Wei Zhou & Alison DeLong

  6. Centre for Organelle Research, University of Stavanger, N-4036, Norway

    Else M. Jonassen & Cathrine Lillo

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Contributions

Z-Y.W. and Y.S. conceived the project and designed the experiments. C.W., M.Y. and Y.S. carried out BZR1 complex purification. Z.D. carried out in-gel digestion, and J.A.O-P. and A.L.B. did the mass spectrometry analysis (Supplementary Fig. S2). W.T. contributed data in Figs 1b,d, 2a–g, 3d, 4d,e,h and Supplementary Figs S4 and S5. M.Y. contributed data in Figs 1c, 2e, 3a–c,e, 4b,c,g, 5b,c and Supplementary Fig. S8. R.W. made contributions to Figs 1a,b, 2a–g, 3d, 4d,e,h and Supplementary Figs S4 and S5. Y.Y. contributed to Fig. 1a, Supplementary Fig. S3, Fig. 4b and Supplementary Fig. S7. T-W.K. contributed Fig. 5a. H-W.Z. and A.D.L. provided the YFP fusions to RCN1 and PP2AA3, the rcn1 pp2aa3 mutant and the anti-PP2AA antibodies. S.S.G. and J.M.G. identified the bzs247 and bzs248 mutants (Fig. 4f). E.M.J. and C.L. provided a p p2a b′αβ mutant and qRT-PCR data (Supplementary Fig. S6). Z-Y.W. and Y.S. wrote the paper together with W.T., M.Y. and R.W.

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Correspondence to Ying Sun or Zhi-Yong Wang.

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Tang, W., Yuan, M., Wang, R. et al. PP2A activates brassinosteroid-responsive gene expression and plant growth by dephosphorylating BZR1. Nat Cell Biol 13, 124–131 (2011). https://doi.org/10.1038/ncb2151

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