title: Fast-forward genetics identifies plant CPL phosphatases as regulators of miRNA processing factor HYL1.
abstract: MicroRNAs (miRNAs) are processed from primary transcripts that contain partially self-complementary foldbacks. As in animals, the core microprocessor in plants is a Dicer protein, DICER-LIKE1 (DCL1). Processing accuracy and strand selection is greatly enhanced through the RNA binding protein HYPONASTIC LEAVES 1 (HYL1) and the zinc finger protein SERRATE (SE). We have combined a luciferase-based genetic screen with whole-genome sequencing for rapid identification of new regulators of miRNA biogenesis and action. Among the first six mutants analyzed were three alleles of C-TERMINAL DOMAIN PHOSPHATASE-LIKE 1 (CPL1)/FIERY2 (FRY2). In the miRNA processing complex, SE functions as a scaffold to mediate CPL1 interaction with HYL1, which needs to be dephosphorylated for optimal activity. In the absence of CPL1, HYL1 dephosphorylation and hence accurate processing and strand selection from miRNA duplexes are compromised. Our findings thus define a new regulatory step in plant miRNA biogenesis.
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany.
authors.forename: Pablo A
authors.initials: PA
authors.lastname: Manavella
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Hagmann
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany.
authors.forename: Felix
authors.initials: F
authors.lastname: Ott
authors.affiliation: Center for Plant Molecular Biology (ZMBP), University of Tübingen, 72076 Tübingen, Germany.
authors.forename: Sascha
authors.initials: S
authors.lastname: Laubinger
authors.affiliation: Proteome Center, Interfaculty Institute for Cell Biology, University of Tübingen, 72076 Tübingen, Germany.
authors.forename: Mirita
authors.initials: M
authors.lastname: Franz
authors.affiliation: Proteome Center, Interfaculty Institute for Cell Biology, University of Tübingen, 72076 Tübingen, Germany.
authors.forename: Boris
authors.initials: B
authors.lastname: Macek
authors.affiliation: Department of Molecular Biology, Max Planck Institute for Developmental Biology, 72076 Tübingen, Germany. Electronic address: weigel@weigelworld.org.
authors.forename: Detlef
authors.initials: D
authors.lastname: Weigel
mesh_terms: D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002135:Calcium-Binding Proteins / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000378:metabolism*; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D010766:Phosphorylation; D018515:Plant Leaves / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D018749:RNA, Plant / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D000072099:Serrate-Jagged Proteins; D014026:Nicotiana / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D002135:Calcium-Binding Proteins; C420966:HYL1 protein, Arabidopsis; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D035683:MicroRNAs; D010940:Plant Proteins; D018749:RNA, Plant; D016601:RNA-Binding Proteins; C000606277:SE protein, Arabidopsis; D000072099:Serrate-Jagged Proteins; D014157:Transcription Factors; C465438:CPL1 protein, Arabidopsis; D010749:Phosphoprotein Phosphatases