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- EMDB-21687: CryoEM structure of the SLC38A9-RagA-RagC-Ragulator complex in th... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-21687 | |||||||||
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Title | CryoEM structure of the SLC38A9-RagA-RagC-Ragulator complex in the post-GAP state | |||||||||
![]() | Ragulator-RagA-GDP-RagC-XDP-SLC38A9; density modified with LocScale | |||||||||
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![]() | small GTPase / mTORC1 activation / amino acid signaling / lysosome / SIGNALING PROTEIN | |||||||||
Function / homology | ![]() asparagine transport / L-asparagine transmembrane transporter activity / sterol sensor activity / L-arginine transmembrane transporter activity / regulation of cholesterol import / L-arginine transmembrane transport / positive regulation of protein localization to lysosome / regulation of cell-substrate junction organization / Gtr1-Gtr2 GTPase complex / regulation of cholesterol efflux ...asparagine transport / L-asparagine transmembrane transporter activity / sterol sensor activity / L-arginine transmembrane transporter activity / regulation of cholesterol import / L-arginine transmembrane transport / positive regulation of protein localization to lysosome / regulation of cell-substrate junction organization / Gtr1-Gtr2 GTPase complex / regulation of cholesterol efflux / L-glutamine transmembrane transporter activity / FNIP-folliculin RagC/D GAP / Ragulator complex / glutamine transport / protein localization to cell junction / amino acid transmembrane transport / L-leucine transmembrane transporter activity / regulation of TORC1 signaling / L-amino acid transmembrane transporter activity / protein localization to lysosome / regulation of TOR signaling / fibroblast migration / MTOR signalling / Amino acids regulate mTORC1 / lysosome localization / endosome organization / Energy dependent regulation of mTOR by LKB1-AMPK / TORC1 signaling / amino acid transmembrane transporter activity / kinase activator activity / arginine binding / protein localization to membrane / azurophil granule membrane / endosomal transport / lysosome organization / cholesterol binding / small GTPase-mediated signal transduction / Macroautophagy / regulation of cell size / RHOJ GTPase cycle / RHOQ GTPase cycle / mTORC1-mediated signalling / tertiary granule membrane / CDC42 GTPase cycle / RHOH GTPase cycle / ficolin-1-rich granule membrane / RHOG GTPase cycle / regulation of receptor recycling / positive regulation of TOR signaling / RAC2 GTPase cycle / RAC3 GTPase cycle / enzyme-substrate adaptor activity / response to amino acid / cellular response to nutrient levels / specific granule membrane / protein-membrane adaptor activity / RAC1 GTPase cycle / positive regulation of TORC1 signaling / negative regulation of autophagy / RNA splicing / cellular response to amino acid starvation / viral genome replication / cholesterol homeostasis / guanyl-nucleotide exchange factor activity / cellular response to starvation / Regulation of PTEN gene transcription / tumor necrosis factor-mediated signaling pathway / positive regulation of interleukin-8 production / TP53 Regulates Metabolic Genes / cellular response to amino acid stimulus / phosphoprotein binding / regulation of cell growth / MAP2K and MAPK activation / response to virus / positive regulation of protein localization to nucleus / GDP binding / late endosome membrane / E3 ubiquitin ligases ubiquitinate target proteins / intracellular protein localization / late endosome / glucose homeostasis / GTPase binding / molecular adaptor activity / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / endosome membrane / lysosome / positive regulation of canonical NF-kappaB signal transduction / positive regulation of MAPK cascade / intracellular signal transduction / membrane raft / protein heterodimerization activity / lysosomal membrane / focal adhesion / intracellular membrane-bounded organelle / GTPase activity / ubiquitin protein ligase binding / apoptotic process / DNA-templated transcription / Neutrophil degranulation / positive regulation of gene expression Similarity search - Function | |||||||||
Biological species | ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
![]() | Fromm SA / Hurley JH | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural mechanism for amino acid-dependent Rag GTPase nucleotide state switching by SLC38A9. Authors: Simon A Fromm / Rosalie E Lawrence / James H Hurley / ![]() Abstract: The Rag GTPases (Rags) recruit mTORC1 to the lysosomal membrane in response to nutrients, where it is then activated in response to energy and growth factor availability. The lysosomal folliculin ...The Rag GTPases (Rags) recruit mTORC1 to the lysosomal membrane in response to nutrients, where it is then activated in response to energy and growth factor availability. The lysosomal folliculin (FLCN) complex (LFC) consists of the inactive Rag dimer, the pentameric scaffold Ragulator, and the FLCN:FNIP2 (FLCN-interacting protein 2) GTPase activating protein (GAP) complex, and prevents Rag dimer activation during amino acid starvation. How the LFC is disassembled upon amino acid refeeding is an outstanding question. Here we show that the cytoplasmic tail of the human lysosomal solute carrier family 38 member 9 (SLC38A9) destabilizes the LFC and thereby triggers GAP activity of FLCN:FNIP2 toward RagC. We present the cryo-EM structures of Rags in complex with their lysosomal anchor complex Ragulator and the cytoplasmic tail of SLC38A9 in the pre- and post-GTP hydrolysis state of RagC, which explain how SLC38A9 destabilizes the LFC and so promotes Rag dimer activation. | |||||||||
History |
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Structure visualization
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Structure viewer | EM map: ![]() ![]() ![]() |
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Downloads & links
-EMDB archive
Map data | ![]() | 1.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 27 KB 27 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 10.1 KB | Display | ![]() |
Images | ![]() | 112.4 KB | ||
Masks | ![]() ![]() | 91.1 MB 91.1 MB | ![]() | |
Filedesc metadata | ![]() | 7.3 KB | ||
Others | ![]() ![]() | 84.5 MB 84.5 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 1 MB | Display | ![]() |
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Full document | ![]() | 1 MB | Display | |
Data in XML | ![]() | 17.8 KB | Display | |
Data in CIF | ![]() | 22.6 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6wj3MC ![]() 6wj2C C: citing same article ( M: atomic model generated by this map |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Ragulator-RagA-GDP-RagC-XDP-SLC38A9; density modified with LocScale | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.137 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
+Entire : Complex of pentameric Ragulator, dimeric Rag GTPases and SLC38A9
+Supramolecule #1: Complex of pentameric Ragulator, dimeric Rag GTPases and SLC38A9
+Supramolecule #2: pentameric Ragulator
+Supramolecule #3: dimeric Rag GTPases
+Supramolecule #4: SLC38A9
+Macromolecule #1: Ragulator complex protein LAMTOR1
+Macromolecule #2: Ragulator complex protein LAMTOR2
+Macromolecule #3: Ragulator complex protein LAMTOR3
+Macromolecule #4: Ragulator complex protein LAMTOR4
+Macromolecule #5: Ragulator complex protein LAMTOR5
+Macromolecule #6: Ras-related GTP-binding protein A
+Macromolecule #7: Ras-related GTP-binding protein C
+Macromolecule #8: Sodium-coupled neutral amino acid transporter 9
+Macromolecule #9: GUANOSINE-5'-DIPHOSPHATE
+Macromolecule #10: xanthosine diphosphate
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.5 mg/mL |
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Buffer | pH: 7.4 |
Grid | Model: C-flat-2/1 / Material: COPPER / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 45 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Average exposure time: 5.7 sec. / Average electron dose: 60.5 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL |
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Output model | ![]() PDB-6wj3: |