KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit anti-Raptor | EMD Millipore | 09-217 |
| Rabbit anti-HA | Cell Signaling Technology | 3724 |
| Rabbit anti-Flag | Cell Signaling Technology | 14793 |
| Rabbit anti-S6K1 | Cell Signaling Technology | 9202 |
| Rabbit anti-pT389-S6K1 | Cell Signaling Technology | 9205 |
| Rabbit anti-mTOR | Cell Signaling Technology | 2983 |
| Anti-rabbit, HRP-linked antibody | Cell Signaling Technology | 7074 |
| Bacterial Strains | ||
| BL21(DE3) | NEB | C2527 |
| LOBSTR | Kerafast | EC1002 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| HEPES | Sigma-Aldrich | H3375 |
| Potassium acetate | Sigma-Aldrich | P1190 |
| Magnesium chloride | Sigma-Aldrich | M8266 |
| Sodium chloride | Sigma-Aldrich | S7563 |
| DTT | Sigma-Aldrich | D5545 |
| Chaps | Sigma-Aldrich | C5070 |
| Triton | Sigma-Aldrich | T9284 |
| Imidazole | Sigma-Aldrich | I5513 |
| GTP | Sigma-Aldrich | G8877 |
| GDP | Sigma-Aldrich | G7127 |
| TCEP | Sigma-Aldrich | C4706 |
| ATP | Sigma-Aldrich | A7699 |
| α-32P-GTP | PerkinElmer | BLU006H250UC |
| DMEM | Thermal Fisher Scientific | 10564-011 |
| SMM 293-TII | Sinobiological | M293TII |
| Deposited data | ||
| Atomic model for GATOR1-Rag-Ragulator (Inhibitory mode) | Protein Data Bank | 7T3A |
| Atomic model for GATOR1-Rag-Ragulator (GAP mode) | Protein Data Bank | 7T3B |
| Atomic model for GATOR1-Rag-Ragulator (Dual mode) | Protein Data Bank | 7T3C |
| EM density map for GATOR1-Rag-Ragulator (Inhibitory mode) | EM Data Bank | 25652 |
| EM density map for GATOR1-Rag-Ragulator (GAP mode) | EM Data Bank | 25653 |
| EM density map for GATOR1-Rag-Ragulator (Dual mode) | EM Data Bank | 25654 |
| Raw images for western blots and Coomassie staining gel | Mendeley | DOI: 10.17632/4vnfng4w4b.1 |
| Experimental Models: Cell Lines | ||
| Human: HEK-293T | ATCC | CRL-3216 |
| Human: FreeStyle 293-F | Thermal Fisher Scientific | R79007 |
| Recombinant DNA | ||
| [pSK109] pETDuet-1 RagA-RagC | Addgene | 99641 |
| [pSK235] pK RagA-RagC | Addgene | 99649 |
| [pSK315] pRK5 Flag-DEPDC5 | Addgene | 46327 |
| [pSK163] pRK5 HA-NPRL2 | Addgene | 99709 |
| [pSK164] pRK5 HA-NPRL3 | Addgene | 46330 |
| [pSK66] pRK5 Flag-S6K1(rat) | Addgene | 100509 |
| Software and Algorithms | ||
| Prism | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| Relion | (Zivanov et al., 2018) | https://www3.mrc-lmb.cam.ac.uk/relion/index.php?title=Main_Page |
| SerialEM | (Mastronarde, 2005) | http://bio3d.colorado.edu/SerialEM/ |
| cryoSPARC | (Punjani et al., 2017) | https://guide.cryosparc.com/ |
| Coot | (Emsley et al., 2010) | https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/ |
| Phenix | (Liebschner et al., 2019) | http://www.phenix-online.org/ |
| Other | ||
| Flag-M2 affinity resin | Sigma-Aldrich | A2220 |
| Ni-NTA affinity resin | EMD Millipore | 30230 |
| Cellulose 300 PEI thin layer chromatography plates | Sorbtech | 1122126 |