RRC ID 3073
著者 Park NI, Muench DG.
タイトル Biochemical and cellular characterization of the plant ortholog of PYM, a protein that interacts with the exon junction complex core proteins Mago and Y14.
ジャーナル Planta
Abstract The exon junction complex (EJC) plays an important role in post-transcriptional control of gene expression. Mago nashi (Mago) and Y14 are core EJC proteins that operate as a functional unit in animal cells, and the Mago-Y14 heterodimer interacts with other EJC core and peripheral proteins. Little is known about the biochemical and cellular characteristics of the EJC and its orthologs in plants. Here, we demonstrate that Arabidopsis Mago and Y14 form a ternary complex with PYM, an RNA-binding protein that was previously shown to interact with the Mago-Y14 heterodimer in Drosophila. Fluorescence microscopy indicated that Arabidopsis Mago and Y14 are localized primarily in the nucleus, whereas PYM is mostly cytoplasmic. In vitro pull-down assays using recombinant proteins showed that the amino-terminal region of the Arabidopsis PYM interacts with the Mago-Y14 heterodimer, a similar observation to that previously reported for the animal versions of these proteins. However, we demonstrated also that Arabidopsis PYM has the ability to interact with monomeric Mago and monomeric Y14. Immunoprecipitation and tandem affinity purification from whole cell extracts detected a subtle interaction between the Arabidopsis Mago-Y14 heterodimer and PYM in flowers, indicating that the ternary complex is not abundant in plant cells. The regions of the polypeptide responsible for nuclear import and export were defined using protein truncations and site-directed mutagenesis. This study identifies unique characteristics of Arabidopsis Mago, Y14 and PYM compared to those observed in animal cells. These are predicted to have important functional implications associated with post-transcriptional regulation of gene expression in plant cells.
巻・号 225(3)
ページ 625-39
公開日 2007-2-1
DOI 10.1007/s00425-006-0385-y
PMID 16953428
MeSH Amino Acid Sequence Animals Arabidopsis Proteins / chemistry Arabidopsis Proteins / genetics Arabidopsis Proteins / metabolism* Cell Nucleus / metabolism Cytosol / metabolism Dimerization Drosophila Proteins / genetics Drosophila Proteins / metabolism Glutathione Transferase / genetics Glutathione Transferase / metabolism Humans Immunoblotting Immunoprecipitation Microscopy, Fluorescence Molecular Sequence Data Nuclear Proteins / chemistry Nuclear Proteins / genetics Nuclear Proteins / metabolism* Plants, Genetically Modified Protein Binding RNA-Binding Proteins / chemistry RNA-Binding Proteins / genetics RNA-Binding Proteins / metabolism* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Sequence Alignment
IF 3.39
引用数 26
WOS 分野 PLANT SCIENCES
リソース情報
シロイヌナズナ / 植物培養細胞・遺伝子 pda02973 pda12755