title: The EJC binding and dissociating activity of PYM is regulated in Drosophila.
abstract: In eukaryotes, RNA processing events in the nucleus influence the fate of transcripts in the cytoplasm. The multi-protein exon junction complex (EJC) associates with mRNAs concomitant with splicing in the nucleus and plays important roles in export, translation, surveillance and localization of mRNAs in the cytoplasm. In mammalian cells, the ribosome associated protein PYM (HsPYM) binds the Y14-Mago heterodimer moiety of the EJC core, and disassembles EJCs, presumably during the pioneer round of translation. However, the significance of the association of the EJC with mRNAs in a physiological context has not been tested and the function of PYM in vivo remains unknown. Here we address PYM function in Drosophila, where the EJC core proteins are genetically required for oskar mRNA localization during oogenesis. We provide evidence that the EJC binds oskar mRNA in vivo. Using an in vivo transgenic approach, we show that elevated amounts of the Drosophila PYM (DmPYM) N-terminus during oogenesis cause dissociation of EJCs from oskar RNA, resulting in its mislocalization and consequent female sterility. We find that, in contrast to HsPYM, DmPYM does not interact with the small ribosomal subunit and dismantles EJCs in a translation-independent manner upon over-expression. Biochemical analysis shows that formation of the PYM-Y14-Mago ternary complex is modulated by the PYM C-terminus revealing that DmPYM function is regulated in vivo. Furthermore, we find that whereas under normal conditions DmPYM is dispensable, its loss of function is lethal to flies with reduced y14 or mago gene dosage. Our analysis demonstrates that the amount of DmPYM relative to the EJC proteins is critical for viability and fertility. This, together with the fact that the EJC-disassembly activity of DmPYM is regulated, implicates PYM as an effector of EJC homeostasis in vivo.
authors: European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Sanjay
authors: S
authors: Ghosh
authors: European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Ales
authors: A
authors: Obrdlik
authors: European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Virginie
authors: V
authors: Marchand
authors: European Molecular Biology Laboratory, Heidelberg, Germany.
authors: Anne
authors: A
authors: Ephrussi
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D003593:Cytoplasm; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster; D005091:Exons / Q000235:genetics; D005260:Female; D007247:Infertility, Female / Q000235:genetics; D046912:Multiprotein Complexes / Q000235:genetics* / Q000378:metabolism; D009865:Oocytes / Q000254:growth & development / Q000378:metabolism; D009866:Oogenesis / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D046912:Multiprotein Complexes; C493333:PYM protein, Drosophila; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C091675:osk protein, Drosophila