title: The Small Molecule Hyperphyllin Enhances Leaf Formation Rate and Mimics Shoot Meristem Integrity Defects Associated with AMP1 Deficiency.
abstract: ALTERED MERISTEM PROGRAM1 (AMP1) is a member of the M28 family of carboxypeptidases with a pivotal role in plant development and stress adaptation. Its most prominent mutant defect is a unique hypertrophic shoot phenotype combining a strongly increased organ formation rate with enhanced meristem size and the formation of ectopic meristem poles. However, so far the role of AMP1 in shoot development could not be assigned to a specific molecular pathway nor is its biochemical function resolved. In this work we evaluated the level of functional conservation between AMP1 and its human homolog HsGCPII, a tumor marker of medical interest. We show that HsGCPII cannot substitute AMP1 in planta and that an HsGCPII-specific inhibitor does not evoke amp1-specific phenotypes. We used a chemical genetic approach to identify the drug hyperphyllin (HP), which specifically mimics the shoot defects of amp1, including plastochron reduction and enlargement and multiplication of the shoot meristem. We assessed the structural requirements of HP activity and excluded that it is a cytokinin analog. HP-treated wild-type plants showed amp1-related tissue-specific changes of various marker genes and a significant transcriptomic overlap with the mutant. HP was ineffective in amp1 and elevated the protein levels of PHAVOLUTA, consistent with the postulated role of AMP1 in miRNA-controlled translation, further supporting an AMP1-related mode of action. Our work suggests that plant and animal members of the M28 family of proteases adopted unrelated functions. With HP we provide a tool to characterize the plant-specific functions of this important class of proteins.
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Olena
authors.initials: O
authors.lastname: Poretska
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Saiqi
authors.initials: S
authors.lastname: Yang
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Delphine
authors.identifier: 0000-0002-5480-9740
authors.initials: D
authors.lastname: Pitorre
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Wilfried
authors.initials: W
authors.lastname: Rozhon
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Karin
authors.initials: K
authors.lastname: Zwerger
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Marcos Castellanos
authors.initials: MC
authors.lastname: Uribe
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Sean
authors.initials: S
authors.lastname: May
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Peter
authors.initials: P
authors.lastname: McCourt
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.).
authors.forename: Brigitte
authors.identifier: 0000-0003-1020-0500
authors.initials: B
authors.lastname: Poppenberger
authors.affiliation: Plant Growth Regulation, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (O.P., S.Y., T.S.); Department for Microbiology, Immunobiology and Genetics, Max F. Perutz Laboratories, University of Vienna, A-1030 Vienna, Austria (O.P., D.P., K.Z., T.S.); Biotechnology of Horticultural Crops, TUM School of Life Sciences Weihenstephan, Technische Universität München, D-85354 Freising, Germany (W.R., B.P.); Nottingham Arabidopsis Stock Centre, University of Nottingham, School of Biosciences, Sutton Bonington, LE12 5RD, UK (M.C.U., S.M.); and Cell and Systems Biology, University of Toronto, Toronto ON M5S 3B2, Canada (P.M.) tobias.sieberer@wzw.tum.de.
authors.forename: Tobias
authors.identifier: 0000-0002-4462-9260
authors.initials: T
authors.lastname: Sieberer
mesh_terms: D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001549:Benzamides / Q000737:chemistry / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D002268:Carboxypeptidases / Q000172:deficiency* / Q000378:metabolism*; D017124:Conserved Sequence; D003583:Cytokinins; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006801:Humans; D018519:Meristem / Q000187:drug effects / Q000502:physiology*; D035683:MicroRNAs / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D018515:Plant Leaves / Q000187:drug effects* / Q000254:growth & development; D036226:Seedlings / Q000187:drug effects / Q000235:genetics / Q000648:ultrastructure; D017386:Sequence Homology, Amino Acid; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D059467:Transcriptome / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D001549:Benzamides; D015415:Biomarkers; D003583:Cytokinins; D035683:MicroRNAs; D054852:Small Molecule Libraries; C000622656:hyperphyllin; D002268:Carboxypeptidases; C506681:AMP1 protein, Arabidopsis