title: Arabidopsis miR171-targeted scarecrow-like proteins bind to GT cis-elements and mediate gibberellin-regulated chlorophyll biosynthesis under light conditions.
abstract: An extraordinarily precise regulation of chlorophyll biosynthesis is essential for plant growth and development. However, our knowledge on the complex regulatory mechanisms of chlorophyll biosynthesis is very limited. Previous studies have demonstrated that miR171-targeted scarecrow-like proteins (SCL6/22/27) negatively regulate chlorophyll biosynthesis via an unknown mechanism. Here we showed that SCLs inhibit the expression of the key gene encoding protochlorophyllide oxidoreductase (POR) in light-grown plants, but have no significant effect on protochlorophyllide biosynthesis in etiolated seedlings. Histochemical analysis of β-glucuronidase (GUS) activity in transgenic plants expressing pSCL27::rSCL27-GUS revealed that SCL27-GUS accumulates at high levels and suppresses chlorophyll biosynthesis at the leaf basal proliferation region during leaf development. Transient gene expression assays showed that the promoter activity of PORC is indeed regulated by SCL27. Consistently, chromatin immunoprecipitation and quantitative PCR assays showed that SCL27 binds to the promoter region of PORC in vivo. An electrophoretic mobility shift assay revealed that SCL27 is directly interacted with G(A/G)(A/T)AA(A/T)GT cis-elements of the PORC promoter. Furthermore, genetic analysis showed that gibberellin (GA)-regulated chlorophyll biosynthesis is mediated, at least in part, by SCLs. We demonstrated that SCL27 interacts with DELLA proteins in vitro and in vivo by yeast-two-hybrid and coimmunoprecipitation analysis and found that their interaction reduces the binding activity of SCL27 to the PORC promoter. Additionally, we showed that SCL27 activates MIR171 gene expression, forming a feedback regulatory loop. Taken together, our data suggest that the miR171-SCL module is critical for mediating GA-DELLA signaling in the coordinate regulation of chlorophyll biosynthesis and leaf growth in light.
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China; University of the Chinese Academy of Sciences, Beijing, China.
authors.forename: Zhaoxue
authors.initials: Z
authors.lastname: Ma
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China; University of the Chinese Academy of Sciences, Beijing, China.
authors.forename: Xupeng
authors.initials: X
authors.lastname: Hu
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Cai
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Genetic Engineering, Institute of Plant Biology, Department of Biochemistry, School of Life Sciences, Fudan University, Shanghai, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhou
authors.affiliation: School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Luo
authors.affiliation: School of Life Sciences and Biotechnology, Shanghai Jiaotong University, Shanghai, China.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Yang
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Wang
authors.affiliation: National Key Laboratory of Plant Molecular Genetics, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors.forename: Jirong
authors.initials: J
authors.lastname: Huang
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development; D029681:Arabidopsis Proteins / Q000096:biosynthesis* / Q000235:genetics*; D002734:Chlorophyll / Q000096:biosynthesis*; D018506:Gene Expression Regulation, Plant; D005875:Gibberellins / Q000378:metabolism; D005966:Glucuronidase / Q000096:biosynthesis / Q000235:genetics; D008027:Light; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics; D063245:Plant Development / Q000235:genetics*; D018515:Plant Leaves / Q000235:genetics / Q000254:growth & development; D030821:Plants, Genetically Modified / Q000235:genetics / Q000254:growth & development; D036226:Seedlings / Q000235:genetics / Q000254:growth & development
chemical_list: D029681:Arabidopsis Proteins; C109555:GAI protein, Arabidopsis; D005875:Gibberellins; C556250:MIRN171 microRNA, Arabidopsis; D035683:MicroRNAs; D008869:Microtubule-Associated Proteins; C472495:POR protein, Arabidopsis; C100978:SCR protein, Arabidopsis; D002734:Chlorophyll; D005966:Glucuronidase
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