title: Triplin, a small molecule, reveals copper ion transport in ethylene signaling from ATX1 to RAN1.
abstract: Copper ions play an important role in ethylene receptor biogenesis and proper function. The copper transporter RESPONSIVE-TO-ANTAGONIST1 (RAN1) is essential for copper ion transport in Arabidopsis thaliana. However it is still unclear how copper ions are delivered to RAN1 and how copper ions affect ethylene receptors. There is not a specific copper chelator which could be used to explore these questions. Here, by chemical genetics, we identified a novel small molecule, triplin, which could cause a triple response phenotype on dark-grown Arabidopsis seedlings through ethylene signaling pathway. ran1-1 and ran1-2 are hypersensitive to triplin. Adding copper ions in growth medium could partially restore the phenotype on plant caused by triplin. Mass spectrometry analysis showed that triplin could bind copper ion. Compared to the known chelators, triplin acts more specifically to copper ion and it suppresses the toxic effects of excess copper ions on plant root growth. We further showed that mutants of ANTIOXIDANT PROTEIN1 (ATX1) are hypersensitive to tiplin, but with less sensitivity comparing with the ones of ran1-1 and ran1-2. Our study provided genetic evidence for the first time that, copper ions necessary for ethylene receptor biogenesis and signaling are transported from ATX1 to RAN1. Considering that triplin could chelate copper ions in Arabidopsis, and copper ions are essential for plant and animal, we believe that, triplin not only could be useful for studying copper ion transport of plants, but also could be useful for copper metabolism study in animal and human.
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|University of Chinese Academy of Sciences, Shanghai, China.
authors.forename: Wenbo
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee, United States of America.
authors.forename: Randy F
authors.identifier: 0000-0002-7585-2238
authors.initials: RF
authors.lastname: Lacey
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|University of Chinese Academy of Sciences, Shanghai, China.
authors.forename: Yajin
authors.initials: Y
authors.lastname: Ye
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|University of Chinese Academy of Sciences, Shanghai, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Lu
authors.affiliation: Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.
authors.forename: Kuo-Chen
authors.initials: KC
authors.lastname: Yeh
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Youli
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Shanghai, China.
authors.forename: Laigeng
authors.initials: L
authors.lastname: Li
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Shanghai, China.
authors.forename: Chi-Kuang
authors.identifier: 0000-0002-5345-0756
authors.initials: CK
authors.lastname: Wen
authors.affiliation: Department of Biochemistry and Cellular and Molecular Biology, University of Tennessee, Knoxville, Tennessee, United States of America.
authors.forename: Brad M
authors.initials: BM
authors.lastname: Binder
authors.affiliation: Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.|Faculty of Life Science and Technology, Kunming University of Science and Technology, 68 Wenchang Road, Yunnan, China.
authors.forename: Yang
authors.identifier: 0000-0001-8697-5148
authors.initials: Y
authors.lastname: Zhao
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism; D003300:Copper / Q000378:metabolism*; D000079922:Copper Transport Proteins; D005030:Ethylenes / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D017136:Ion Transport / Q000235:genetics; D063245:Plant Development; D030821:Plants, Genetically Modified; D016601:RNA-Binding Proteins; D036226:Seedlings / Q000235:genetics; D015398:Signal Transduction; D013890:Thiourea / Q000031:analogs & derivatives; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D020931:ran GTP-Binding Protein
chemical_list: D029681:Arabidopsis Proteins; C490178:At5g20010 protein, Arabidopsis; D027682:Cation Transport Proteins; D000079922:Copper Transport Proteins; D005030:Ethylenes; D016601:RNA-Binding Proteins; D014157:Transcription Factors; C000619665:triplin; D003300:Copper; C036216:ethylene; C504056:At2g31650 protein, Arabidopsis; D011495:Histone-Lysine N-Methyltransferase; D020931:ran GTP-Binding Protein; D013890:Thiourea