Abstract
Peptide-mediated cell-to-cell signaling has crucial roles in coordination and definition of cellular functions in plants. Peptide-receptor matching is important for understanding the mechanisms underlying peptide-mediated signaling. Here we report the structure-guided identification of root meristem growth factor (RGF) receptors important for plant development. An assay based on a signature ligand recognition motif (Arg-x-Arg) conserved in a subfamily of leucine-rich repeat receptor kinases (LRR-RKs) identified the functionally uncharacterized LRR-RK At4g26540 as a receptor of RGF1 (RGFR1). We further solved the crystal structure of RGF1 in complex with the LRR domain of RGFR1 at a resolution of 2.6 Å, which reveals that the Arg-x-Gly-Gly (RxGG) motif is responsible for specific recognition of the sulfate group of RGF1 by RGFR1. Based on the RxGG motif, we identified additional four RGFRs. Participation of the five RGFRs in RGF-induced signaling is supported by biochemical and genetic data. We also offer evidence showing that SERKs function as co-receptors for RGFs. Taken together, our study identifies RGF receptors and co-receptors that can link RGF signals with their downstream components and provides a proof of principle for structure-based matching of LRR-RKs with their peptide ligands.
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Acknowledgements
We thank S Huang and J He at Shanghai Synchrotron Radiation Facility (SSRF) for assistance with data collection; H Deng and J Liu at the center for Biomedical Analysis of Tsinghua University for MS analysis; L Li and A Jiang at Peking University for providing the incubator. We acknowledge the Tsinghua University Branch of China National Center for Protein Sciences Beijing for providing the facility support. This work was funded by Projects of International Cooperation and Exchanges of the National Natural Science Foundation of China (31420103906), the Ministry of Science and Technology of China (2015CB910200) and State Key Program of the National Natural Science of China (31130063 and 31421001) to JC; the National Natural Science Foundation of China (91217305 and 91017010) to HG.
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Identification of the LRR-RK At4g26540 as a receptor of RGF1 by Gel-filtration and MS. (PDF 100 kb)
Supplementary information, Figure S2
Quantitative assays of RGFR1LRR interaction with RGFs. (PDF 246 kb)
Supplementary information, Figure S3
Conserved recognition mechanism of RGFs by RGFR1. (PDF 327 kb)
Supplementary information, Figure S4
Sequence alignment the extracellular LRR domains of RGFR1-5. (PDF 702 kb)
Supplementary information, Figure S5
RGF1 interacts with RGFR2LRR in vitro. (PDF 75 kb)
Supplementary information, Figure S6
RGFR1/2 expression patterns in root. (PDF 318 kb)
Supplementary information, Figure S7
Expression patterns of RGFs and RGFRs. (PDF 272 kb)
Supplementary information, Figure S8
Schematic map of the T-DNA insertion mutants of rgfr1, rgfr2, rgfr3, rgfr4 and rgfr5. (PDF 147 kb)
Supplementary information, Figure S9
Dose-dependent RGF1 response pattern of Col-0. (PDF 92 kb)
Supplementary information, Figure S10
RGF1 induced meristem response in rgfr mutants. (PDF 110 kb)
Supplementary information, Figure S11
RGF1 binding induces no oligomerization of RGFR1LRR in vitro. (PDF 103 kb)
Supplementary information, Figure S12
RGF1 induces RGFR1LRR-SERKs heterodimerization. (PDF 152 kb)
Supplementary information, Figure S13
RGF1-5, 8 but not RGF6, 7, 9 induce RGFR1LRR-SERK1LRR heterodimerization in vitro. (PDF 260 kb)
Supplementary information, Figure S14
The RxR motif are conserved in different plant species. (PDF 461 kb)
Supplementary information, Table S1
Peptide list. (PDF 359 kb)
Supplementary information, Table S2
Summary of crystallography analysis. (PDF 389 kb)
Supplementary information, Table S3
Primer list. (PDF 116 kb)
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Song, W., Liu, L., Wang, J. et al. Signature motif-guided identification of receptors for peptide hormones essential for root meristem growth. Cell Res 26, 674–685 (2016). https://doi.org/10.1038/cr.2016.62
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DOI: https://doi.org/10.1038/cr.2016.62
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