Translation driven by an eIF4G core domain in vivo.
Most eukaryotic mRNAs possess a 5' cap structure (m(7)GpppN) and a 3' poly(A) tail which promote translation initiation by binding the eukaryotic translation initiation factor (eIF)4E and the poly(A) binding protein (PABP), respectively. eIF4G can bridge between eIF4E and PABP, and-through eIF3-is thought to establish a link to the small ribosomal subunit. We fused the C-terminal region of human eIF4GI lacking both the eIF4E- and PABP-binding sites, to the IRE binding protein IRP-1. This chimeric protein suffices to direct the translation of the downstream cistron of bicistronic mRNAs bearing IREs in their intercistronic space in vivo. This function is preserved even when translation via the 5' end is inhibited. Deletion analysis defined the conserved central domain (amino acids 642-1091) of eIF4G as an autonomous 'ribosome recruitment core' and implicated eIF4A as a critical binding partner. Our data reveal the sufficiency of the conserved eIF4G ribosome recruitment core to drive productive mRNA translation in living cells. The C-terminal third of eIF4G is dispensable, and may serve as a regulatory domain.
De Gregorio
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Gene Expression Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany.
Preiss
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Hentze
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D004305:Dose-Response Relationship, Drug; D039603:Eukaryotic Initiation Factor-4G; D005786:Gene Expression Regulation; D005796:Genes / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000235:genetics*; D010957:Plasmids / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D039603:Eukaryotic Initiation Factor-4G; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins