title: Antagonistic peptide technology for functional dissection of CLV3/ESR genes in Arabidopsis.
abstract: In recent years, peptide hormones have been recognized as important signal molecules in plants. Genetic characterization of such peptides is challenging since they are usually encoded by small genes. As a proof of concept, we used the well-characterized stem cell-restricting CLAVATA3 (CLV3) to develop an antagonistic peptide technology by transformations of wild-type Arabidopsis (Arabidopsis thaliana) with constructs carrying the full-length CLV3 with every residue in the peptide-coding region replaced, one at a time, by alanine. Analyses of transgenic plants allowed us to identify one line exhibiting a dominant-negative clv3-like phenotype, with enlarged shoot apical meristems and increased numbers of floral organs. We then performed second dimensional amino acid substitutions to replace the glycine residue individually with the other 18 possible proteinaceous amino acids. Examination of transgenic plants showed that a glycine-to-threonine substitution gave the strongest antagonistic effect in the wild type, in which over 70% of transgenic lines showed the clv3-like phenotype. Among these substitutions, a negative correlation was observed between the antagonistic effects in the wild type and the complementation efficiencies in clv3. We also demonstrated that such an antagonistic peptide technology is applicable to other CLV3/EMBRYO SURROUNDING REGION (CLE) genes, CLE8 and CLE22, as well as in vitro treatments. We believe this technology provides a powerful tool for functional dissection of widely occurring CLE genes in plants.
authors: Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
authors: Xiu-Fen
authors: XF
authors: Song
authors: Peng
authors: P
authors: Guo
authors: Shi-Chao
authors: SC
authors: Ren
authors: Ting-Ting
authors: TT
authors: Xu
authors: Chun-Ming
authors: CM
authors: Liu
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D017360:Arabidopsis / Q000196:embryology / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005799:Genes, Dominant; D017343:Genes, Plant / Q000235:genetics*; D005816:Genetic Complementation Test; D005966:Glucuronidase / Q000378:metabolism; D018519:Meristem / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D010455:Peptides / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D010641:Phenotype; D030821:Plants, Genetically Modified; D015202:Protein Engineering / Q000379:methods*; D012639:Seeds / Q000378:metabolism
chemical_list: C118167:AT2G27250 protein, Arabidopsis; D029681:Arabidopsis Proteins; D010455:Peptides; D005966:Glucuronidase