Abstract
PYR1/PYL/RCAR family proteins (PYLs) are well-characterized abscisic acid (ABA) receptors. Among the 14 PYL members in Arabidopsis thaliana, PYL13 is ABA irresponsive and its function has remained elusive. Here, we show that PYL13 selectively inhibits the phosphatase activity of PP2CA independent of ABA. The crystal structure of PYL13-PP2CA complex, which was determined at 2.4 Å resolution, elucidates the molecular basis for the specific recognition between PP2CA and PYL13. In addition to the canonical interactions between PYLs and PP2Cs, an extra interface is identified involving an element in the vicinity of a previously uncharacterized CCCH zinc-finger (ZF) motif in PP2CA. Sequence blast identified another 56 ZF-containing PP2Cs, all of which are from plants. The structure also reveals the molecular determinants for the ABA irresponsiveness of PYL13. Finally, biochemical analysis suggests that PYL13 may hetero-oligomerize with PYL10. These two PYLs antagonize each other in their respective ABA-independent inhibitions of PP2Cs. The biochemical and structural studies provide important insights into the function of PYL13 in the stress response of plant and set up a foundation for future biotechnological applications of PYL13.
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Acknowledgements
We thank J He and S Huang at Shanghai Synchrotron Radiation Facility (SSRF) for assistance. This work was supported by funds from the Ministry of Science and Technology (2011CB910501), the National Natural Science Foundation of China (31125009 and 91017011), and Tsinghua University. The research of Nieng Yan was supported in part by an International Early Career Scientist grant from the Howard Hughes Medical Institute.
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
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Supplementary information, Figure S1
Sequence comparison of five Clade A PP2Cs from Arabidopsis thaliana. (PDF 581 kb)
Supplementary information, Figure S2
Sequence comparison of PP2CA with HAI1/2/3. (PDF 369 kb)
Supplementary information, Figure S3
Evolutionary relationships of ZF (zinc-finger motif)-containing PP2Cs. (PDF 384 kb)
Supplementary information, Figure S4
Structure-guided point mutations converted PYL13 into an ABA receptor. (PDF 92 kb)
Supplementary information, Figure S5
PYL13 and PYL10 form hetero-complex. (PDF 37 kb)
Supplementary information Table S1
Statistics of data collection and refinement (PDF 30 kb)
Supplementary information Table S2
ICP-AES (Inductively Coupled Plasma-Atomic Emission Spectrometry) analysis of the metal contents in PP2CA-PYL13 complex and PYL5 (PDF 34 kb)
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Li, W., Wang, L., Sheng, X. et al. Molecular basis for the selective and ABA-independent inhibition of PP2CA by PYL13. Cell Res 23, 1369–1379 (2013). https://doi.org/10.1038/cr.2013.143
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DOI: https://doi.org/10.1038/cr.2013.143
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