eIF4G dramatically enhances the binding of eIF4E to the mRNA 5'-cap structure.
The cap structure, m7GpppN, is present at the 5'-end of all eukaryotic cellular (except organellar) mRNAs. Initiation of translation is mediated by the multisubunit initiation factor eIF4F, which binds the cap structure via its eIF4E subunit and facilitates the binding of mRNA to ribosomes. Here, we used recombinant proteins to reconstitute the cap recognition activity of eIF4F in vitro. We demonstrate that the interaction of eIF4E with the mRNA 5'-cap structure is dramatically enhanced by eIF4G, as determined by a UV-induced cross-linking assay. Furthermore, assembly of the eIF4F complex at the cap structure, as well as ATP hydrolysis, is shown to be a requisite for the cross-linking of another initiation factor, eIF4B, to the cap structure. In addition, the stimulatory effect of eIF4G on the cap recognition of eIF4E is inhibited by the translational repressor, 4E-BP1. These results suggest that eIF4E initially interacts with the mRNA cap structure as part of the eIF4F complex.
Haghighat
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A
Department of Biochemistry and McGill Cancer Center, Faculty of Medicine, McGill University, Montreal, Quebec, Canada H3G 1Y6.
Sonenberg
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N
D048868:Adaptor Proteins, Signal Transducing; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F; D039603:Eukaryotic Initiation Factor-4G; D039642:Eukaryotic Initiation Factors; D006868:Hydrolysis; D046911:Macromolecular Substances; D051379:Mice; D010448:Peptide Initiation Factors / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism; D010777:Photochemistry; D012315:RNA Caps / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; C499527:Eif4ebp1 protein, mouse; D039561:Eukaryotic Initiation Factor-4E; D039562:Eukaryotic Initiation Factor-4F; D039603:Eukaryotic Initiation Factor-4G; D039642:Eukaryotic Initiation Factors; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D010750:Phosphoproteins; D012315:RNA Caps; D012333:RNA, Messenger; D000255:Adenosine Triphosphate