Supplementary Materials

Structural basis for the docking of mTORC1 on the lysosomal surface

Kacper B. Rogala, Xin Gu, Jibril F. Kedir, Monther Abu-Remaileh, Laura F. Bianchi, Alexia M. S. Bottino, Rikke Dueholm, Anna Niehaus, Daan Overwijn, Ange-Célia Priso Fils, Sherry X. Zhou, Daniel Leary, Nouf N. Laqtom, Edward J. Brignole, David M. Sabatini

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S9
  • Tables S1 and S2
  • Captions for Movies S1 to S5
  • References

Images, Video, and Other Media

Movie S1
The experimental electron density (left) and the corresponding molecular model (right) of the Raptor-Rag-Ragulator complex.
Movie S2
Variability analysis of the Raptor-Rag-Ragulator complex structure. Note the swinging motion of the two halves of the complex.
Movie S3
Following GTP hydrolysis, strand β2 of the RagC interswitch changes its register by two residues, such that the β2-β3 interswitch loop becomes more compact. This interswitch movement has further implications on the position of the GTPase domain in the context of the Rag heterodimer (see Fig. 4C, 4D). Molecular dynamics simulations revealed that the switch machinery of RagC in the GTP-bound state is fairly stable. However, upon hydrolysis to GDP, and following the interswitch shift, a large portion of the RagC switch machinery becomes highly dynamic (switch I, interswitch strand β2 and a part of switch II). Indeed, all those dynamic switch regions are also found disordered in our cryo-EM structure of the Raptor-Rag-Ragulator complex.
Movie S4
Raptor is unable to form a lasting interaction with Rag GTPases in the GTP-GTP nucleotide state. Following GTP hydrolysis by RagC, an extra space is made in between the two GTPase domains of the Rags. The Raptor claw can detect the availability of this space, and therefore by extension - detect the nucleotide state of RagC. Please note that the modeled GTPase domain of RagC•GTP in this movie was not corrected for its interswitch-induced shift relative to the CRD.
Movie S5
Dimeric mTOR forms a large protein assembly through its interaction with Raptor (to form mTORC1), and through Rag-Ragulator that anchors it to the lysosomal surface. Such positioned mTORC1-Rag-Ragulator supercomplex becomes allosterically activated through its association with Rheb GTPases.