Abstract
Strigolactone plant hormones control plant architecture and are key players in both symbiotic and parasitic interactions. They contain an ABC tricyclic lactone connected to a butenolide group, the D ring. The DWARF14 (D14) strigolactone receptor belongs to the superfamily of α/β-hydrolases, and is known to hydrolyze the bond between the ABC lactone and the D ring. Here we characterized the binding and catalytic functions of RAMOSUS3 (RMS3), the pea (Pisum sativum) ortholog of rice (Oryza sativa) D14 strigolactone receptor. Using new profluorescent probes with strigolactone-like bioactivity, we found that RMS3 acts as a single-turnover enzyme that explains its apparent low enzymatic rate. We demonstrated the formation of a covalent RMS3-D-ring complex, essential for bioactivity, in which the D ring was attached to histidine 247 of the catalytic triad. These results reveal an undescribed mechanism of plant hormone reception in which the receptor performs an irreversible enzymatic reaction to generate its own ligand.
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Acknowledgements
We thank R. Novaretti for plant bioassays, A.E. Stewart for helpful discussion, S.K. Lin for technical assistance, J.-P. Andrieu for the assistance and access to amino acids determination facility, U. Pedmale for technical advice, and B.C. Willige, C. Bourbousse, J. Woodson, U. Pedmale, A. Seluzicki and D. O'Keefe for their comments on the manuscript. We are grateful to the Institut National de la Recherche Agronomique (INRA), the Agence Nationale de la Recherche (contract ANR-12-BSV6-004-01) and the Stream COST Action FA1206 for financial support. A.d.S.G. and J.C. were partially supported by a grant to J.C. from US National Institutes of Health (R01 GM094428). J.C. is funded as an investigator of the Howard Hughes Medical Institute. A.d.S.G. was partially supported by a grant from Catharina Foundation to the Salk Institute. The Institut Jean-Pierre Bourgin benefits from the support of the Labex Saclay Plant Sciences-SPS (ANR-10-LABX-0040-SPS).
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A.d.S.G., G.C., J.C., C.R., F.-D.B. designed research; G.C. designed and synthesized the probes; G.C., F.-D.B. synthesized the other chemicals; A.d.S.G., M.-A.B.-D. produced and purified the proteins; A.d.S.G. characterized the proteins; A.d.S.G., G.C. did the kinetic experiments; A.d.S.G., J.-P.P., S.B., C.R., F.-D.B. performed the plant experiments; D.C., J.-P.L.C. performed the mass experiments; G.C., F.P., F.-D.B. performed the HPLC analyses and separations; P.R. did the x-ray analyses; M.B. did the protein modeling; C.T. performed strigolactone quantifications in pea; A.d.S.G., G.C., M.-A.B.-D., J.-P.L.C., P.R., C.T., J.C., S.B., C.R., F.-D.B. analyzed data; A.d.S.G., C.R., F.-D.B. wrote the paper.
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de Saint Germain, A., Clavé, G., Badet-Denisot, MA. et al. An histidine covalent receptor and butenolide complex mediates strigolactone perception. Nat Chem Biol 12, 787–794 (2016). https://doi.org/10.1038/nchembio.2147
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DOI: https://doi.org/10.1038/nchembio.2147
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