原创力文档the hierarchy of exon-junction complex assembly by the spliceosome explains key features of mammalian nonsense-mediated mrna decayexon-junction层次复杂装配哺乳动物nonsense-mediated信使rna剪接体解释了关键特性的衰变.pdfVIP
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the hierarchy of exon-junction complex assembly by the spliceosome explains key features of mammalian nonsense-mediated mrna decayexon-junction层次复杂装配哺乳动物nonsense-mediated信使rna剪接体解释了关键特性的衰变
The Hierarchy of Exon-Junction Complex Assembly by
the Spliceosome Explains Key Features of Mammalian
Nonsense-Mediated mRNA Decay
Niels H. Gehring1,2,3*, Styliani Lamprinaki1,3, Matthias W. Hentze1,3, Andreas E. Kulozik1,2*
1 Molecular Medicine Partnership Unit, University of Heidelberg and European Molecular Biology Laboratory, Heidelberg, Germany, 2 Department of Pediatric Oncology,
Hematology and Immunology, University of Heidelberg, Heidelberg, Germany, 3 European Molecular Biology Laboratory, Heidelberg, Germany
Abstract
Exon junction complexes (EJCs) link nuclear splicing to key features of mRNA function including mRNA stability, translation,
and localization. We analyzed the formation of EJCs by the spliceosome, the physiological EJC assembly machinery. We
studied a comprehensive set of eIF4A3, MAGOH, and BTZ mutants in complete or C-complex–arrested splicing reactions
and identified essential interactions of EJC proteins during and after EJC assembly. These data establish that EJC deposition
proceeds through a defined intermediate, the pre-EJC, as an ordered, sequential process that is coordinated by splicing. The
pre-EJC consists of eIF4A3 and MAGOH-Y14, is formed before exon ligation, and provides a binding platform for peripheral
EJC components that join after release from the spliceosome and connect the core structure with function. Specifically, we
identified BTZ to bridge the EJC to the nonsense-mediated messenger RNA (mRNA) decay protein UPF1, uncovering a
critical link between mRNP architecture and mRNA stability. Based on this systematic analysis of EJC assembly by the
spliceosome, we propose a model of how a functional EJC is assembled in a strictly sequential and hierarchical fashion,
including nuclear splicing-dependent and cytoplasmic steps.
Citation: Gehring NH, Lamprinaki S, Hentze MW, Kulozik AE (2009) The Hierar
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