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Global identification of the Gene‐Specific Association of the Exon‐Exon Junction Complex in Drosophila
Author(s) -
Haque Nazmul,
Blanchette Marco
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.499.5
Subject(s) - exon , rna splicing , biology , rna binding protein , messenger rna , translation (biology) , nonsense mediated decay , gene , rna interference , microbiology and biotechnology , spliceosome , genetics , alternative splicing , rna
Exon Junction Complex (EJC) is a large multi‐protein complex that assembles 20‐24nt upstream of exon‐exon junction following splicing of pre‐mRNAs. EJC is involved in several aspects of mRNA metabolism, including mRNA export, translation and Nonsense Meditated mRNA Decay (NMD). We have recently found in Drosophila that EJCs assemble on specific spliced mRNAs, which prompted us to globally characterize the mRNAs bound by EJCs in Drosophila S2 cells. RNAs bound to eIF4AIII and Y14, two of the core components of EJC, were immunopurified from stable S2 cell lines expressing affinity‐tag version of the proteins and used to hybridize Drosophila whole genome tiling array. We have identified that only ~10% of expressed mRNAs associate with an EJC. Surprisingly, our global analysis have reveal that some mRNAs are bound only by eIF4AIII or Y14, suggesting that these proteins might be part of complexes distinct from EJCs. In addition, global gene expression analysis in eIF4AIII‐depleted S2 cells by RNAi identified an over‐represented population of mRNAs bound by eIF4AIII with reduced expression while no significant overlap was observed in the up‐regulated mRNAs, suggesting a role for eiF4AIII in stabilizing these mRNAs. Bio‐computational analyses are underway to identify cis‐acting elements that could trigger gene‐specific EJC deposition. This project is supported by Stowers Institute for Medical Research