title: Regulation of pri-miRNA processing by the hnRNP-like protein AtGRP7 in Arabidopsis.
abstract: The hnRNP-like glycine-rich RNA-binding protein AtGRP7 regulates pre-mRNA splicing in Arabidopsis. Here we used small RNA-seq to show that AtGRP7 also affects the miRNA inventory. AtGRP7 overexpression caused a significant reduction in the level of 30 miRNAs and an increase for 14 miRNAs with a minimum log2 fold change of ± 0.5. Overaccumulation of several pri-miRNAs including pri-miR398b, pri-miR398c, pri-miR172b, pri-miR159a and pri-miR390 at the expense of the mature miRNAs suggested that AtGRP7 affects pri-miRNA processing. Indeed, RNA immunoprecipitation revealed that AtGRP7 interacts with these pri-miRNAs in vivo. Mutation of an arginine in the RNA recognition motif abrogated in vivo binding and the effect on miRNA and pri-miRNA levels, indicating that AtGRP7 inhibits processing of these pri-miRNAs by direct binding. In contrast, pri-miRNAs of selected miRNAs that were elevated or not changed in response to high AtGRP7 levels were not bound in vivo. Reduced accumulation of miR390, an initiator of trans-acting small interfering RNA (ta-siRNA) formation, also led to lower TAS3 ta-siRNA levels and increased mRNA expression of the target AUXIN RESPONSE FACTOR4. Furthermore, AtGRP7 affected splicing of pri-miR172b and pri-miR162a. Thus, AtGRP7 is an hnRNP-like protein with a role in processing of pri-miRNAs in addition to its role in pre-mRNA splicing.
authors.affiliation: Molecular Cell Physiology, Bielefeld University.
authors.forename: Tino
authors.initials: T
authors.lastname: Köster
authors.affiliation: Molecular Cell Physiology, Bielefeld University.
authors.forename: Katja
authors.initials: K
authors.lastname: Meyer
authors.affiliation: Institute of Computer Science, Martin-Luther-University Halle-Wittenberg, Germany.
authors.forename: Claus
authors.initials: C
authors.lastname: Weinholdt
authors.affiliation: Max Planck Institute for Developmental Biology, Tuebingen, Germany Department of Animal & Plant Sciences, University of Sheffield, UK.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Smith
authors.affiliation: Molecular Cell Physiology, Bielefeld University.
authors.forename: Martina
authors.initials: M
authors.lastname: Lummer
authors.affiliation: Max Planck Institute for Developmental Biology, Tuebingen, Germany Center for Plant Molecular Biology, University of Tuebingen Chemical Genomics Centre of the Max Planck Society, Dortmund, Germany.
authors.forename: Corinna
authors.initials: C
authors.lastname: Speth
authors.affiliation: Institute of Computer Science, Martin-Luther-University Halle-Wittenberg, Germany German Centre for Integrative Biodiversity Research Halle-Jena-Leipzig, Germany.
authors.forename: Ivo
authors.initials: I
authors.lastname: Grosse
authors.affiliation: Max Planck Institute for Developmental Biology, Tuebingen, Germany.
authors.forename: Detlef
authors.initials: D
authors.lastname: Weigel
authors.affiliation: Molecular Cell Physiology, Bielefeld University Institute for Genome Research & Systems Biology, CeBiTec, Bielefeld, Germany dorothee.staiger@uni-bielefeld.de.
authors.forename: Dorothee
authors.initials: D
authors.lastname: Staiger
mesh_terms: D017398:Alternative Splicing; D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D035683:MicroRNAs / Q000378:metabolism*; D018832:Molecular Chaperones / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D034741:RNA, Small Interfering / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D035683:MicroRNAs; D018832:Molecular Chaperones; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D013482:Superoxide Dismutase; C118496:GRP7 protein, Arabidopsis; C534507:GRP8 protein, Arabidopsis; C543882:MIRN398 microRNA, Arabidopsis; C586061:CSD2 protein, Arabidopsis
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