The dynamic mechanism of 4E-BP1 recognition and phosphorylation by mTORC1.
The activation of cap-dependent translation in eukaryotes requires multisite, hierarchical phosphorylation of 4E-BP by the 1 MDa kinase mammalian target of rapamycin complex 1 (mTORC1). To resolve the mechanism of this hierarchical phosphorylation at the atomic level, we monitored by NMR spectroscopy the interaction of intrinsically disordered 4E binding protein ... isoform 1 (4E-BP1) with the mTORC1 subunit regulatory-associated protein of mTOR (Raptor). The N-terminal RAIP motif and the C-terminal TOR signaling (TOS) motif of 4E-BP1 bind separate sites in Raptor, resulting in avidity-based tethering of 4E-BP1. This tethering orients the flexible central region of 4E-BP1 toward the mTORC1 kinase site for phosphorylation. The structural constraints imposed by the two tethering interactions, combined with phosphorylation-induced conformational switching of 4E-BP1, explain the hierarchy of 4E-BP1 phosphorylation by mTORC1. Furthermore, we demonstrate that mTORC1 recognizes both free and eIF4E-bound 4E-BP1, allowing rapid phosphorylation of the entire 4E-BP1 pool and efficient activation of translation. Finally, our findings provide a mechanistic explanation for the differential rapamycin sensitivity of the 4E-BP1 phosphorylation sites.
Mesh Terms:
Adaptor Proteins, Signal Transducing, Binding Sites, Cell Cycle Proteins, Chaetomium, Cloning, Molecular, Crystallography, X-Ray, Escherichia coli, Eukaryotic Initiation Factor-4E, Gene Expression, Genetic Vectors, Humans, Kinetics, Mechanistic Target of Rapamycin Complex 1, Models, Molecular, Phosphorylation, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, Protein Processing, Post-Translational, Recombinant Proteins, Regulatory-Associated Protein of mTOR, Signal Transduction, Structural Homology, Protein, Substrate Specificity, TOR Serine-Threonine Kinases
Adaptor Proteins, Signal Transducing, Binding Sites, Cell Cycle Proteins, Chaetomium, Cloning, Molecular, Crystallography, X-Ray, Escherichia coli, Eukaryotic Initiation Factor-4E, Gene Expression, Genetic Vectors, Humans, Kinetics, Mechanistic Target of Rapamycin Complex 1, Models, Molecular, Phosphorylation, Protein Binding, Protein Conformation, alpha-Helical, Protein Conformation, beta-Strand, Protein Interaction Domains and Motifs, Protein Processing, Post-Translational, Recombinant Proteins, Regulatory-Associated Protein of mTOR, Signal Transduction, Structural Homology, Protein, Substrate Specificity, TOR Serine-Threonine Kinases
Mol Cell
Date: Dec. 03, 2020
PubMed ID: 33852892
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