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Open data
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Basic information
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Title | Cryo-EM structure of the SHOC2:PP1C:MRAS complex | |||||||||
![]() | Final map | |||||||||
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![]() | Phosphatase / leucine rich repeat / RAF / complex / SIGNALING PROTEIN | |||||||||
Function / homology | ![]() cellular response to growth hormone stimulus / protein phosphatase type 1 complex / negative regulation of neural precursor cell proliferation / PTW/PP1 phosphatase complex / regulation of nucleocytoplasmic transport / GTP-dependent protein binding / cyclic-GMP-AMP transmembrane import across plasma membrane / nerve growth factor signaling pathway / protein phosphatase regulator activity / protein phosphatase 1 binding ...cellular response to growth hormone stimulus / protein phosphatase type 1 complex / negative regulation of neural precursor cell proliferation / PTW/PP1 phosphatase complex / regulation of nucleocytoplasmic transport / GTP-dependent protein binding / cyclic-GMP-AMP transmembrane import across plasma membrane / nerve growth factor signaling pathway / protein phosphatase regulator activity / protein phosphatase 1 binding / lamin binding / SHOC2 M1731 mutant abolishes MRAS complex function / Gain-of-function MRAS complexes activate RAF signaling / positive regulation of Ras protein signal transduction / microtubule organizing center / myosin phosphatase activity / protein serine/threonine phosphatase activity / glycogen metabolic process / protein-serine/threonine phosphatase / Triglyceride catabolism / Maturation of hRSV A proteins / entrainment of circadian clock by photoperiod / phosphatase activity / phosphoprotein phosphatase activity / cleavage furrow / blastocyst development / negative regulation of neuron differentiation / fibroblast growth factor receptor signaling pathway / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / positive regulation of glial cell proliferation / Resolution of Sister Chromatid Cohesion / positive regulation of neuron differentiation / protein dephosphorylation / Downregulation of TGF-beta receptor signaling / small monomeric GTPase / G protein activity / cellular response to leukemia inhibitory factor / RHO GTPases Activate Formins / RAF activation / circadian regulation of gene expression / neuron differentiation / regulation of circadian rhythm / kinetochore / positive regulation of neuron projection development / Separation of Sister Chromatids / GDP binding / MAPK cascade / Circadian Clock / presynapse / midbody / actin cytoskeleton organization / spermatogenesis / protein phosphatase binding / mitochondrial outer membrane / Ras protein signal transduction / dendritic spine / nuclear speck / cell cycle / protein domain specific binding / cell division / focal adhesion / GTPase activity / glutamatergic synapse / protein-containing complex binding / nucleolus / GTP binding / protein kinase binding / signal transduction / protein-containing complex / mitochondrion / RNA binding / nucleoplasm / nucleus / metal ion binding / plasma membrane / cytoplasm / cytosol Similarity search - Function | |||||||||
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Method | single particle reconstruction / cryo EM / Resolution: 2.95 Å | |||||||||
![]() | Liau NPD / Johnson MC | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural basis for SHOC2 modulation of RAS signalling. Authors: Nicholas P D Liau / Matthew C Johnson / Saeed Izadi / Luca Gerosa / Michal Hammel / John M Bruning / Timothy J Wendorff / Wilson Phung / Sarah G Hymowitz / Jawahar Sudhamsu / ![]() Abstract: The RAS-RAF pathway is one of the most commonly dysregulated in human cancers. Despite decades of study, understanding of the molecular mechanisms underlying dimerization and activation of the kinase ...The RAS-RAF pathway is one of the most commonly dysregulated in human cancers. Despite decades of study, understanding of the molecular mechanisms underlying dimerization and activation of the kinase RAF remains limited. Recent structures of inactive RAF monomer and active RAF dimer bound to 14-3-3 have revealed the mechanisms by which 14-3-3 stabilizes both RAF conformations via specific phosphoserine residues. Prior to RAF dimerization, the protein phosphatase 1 catalytic subunit (PP1C) must dephosphorylate the N-terminal phosphoserine (NTpS) of RAF to relieve inhibition by 14-3-3, although PP1C in isolation lacks intrinsic substrate selectivity. SHOC2 is as an essential scaffolding protein that engages both PP1C and RAS to dephosphorylate RAF NTpS, but the structure of SHOC2 and the architecture of the presumptive SHOC2-PP1C-RAS complex remain unknown. Here we present a cryo-electron microscopy structure of the SHOC2-PP1C-MRAS complex to an overall resolution of 3 Å, revealing a tripartite molecular architecture in which a crescent-shaped SHOC2 acts as a cradle and brings together PP1C and MRAS. Our work demonstrates the GTP dependence of multiple RAS isoforms for complex formation, delineates the RAS-isoform preference for complex assembly, and uncovers how the SHOC2 scaffold and RAS collectively drive specificity of PP1C for RAF NTpS. Our data indicate that disease-relevant mutations affect complex assembly, reveal the simultaneous requirement of two RAS molecules for RAF activation, and establish rational avenues for discovery of new classes of inhibitors to target this pathway. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 32 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 23.3 KB 23.3 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 8.9 KB | Display | ![]() |
Images | ![]() | 107.9 KB | ||
Filedesc metadata | ![]() | 6.8 KB | ||
Others | ![]() ![]() | 59.4 MB 59.4 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 780.2 KB | Display | ![]() |
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Full document | ![]() | 779.7 KB | Display | |
Data in XML | ![]() | 16.4 KB | Display | |
Data in CIF | ![]() | 21 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 7sd0MC ![]() 7sd1C M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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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 | Final map | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.838 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Half map A
File | emd_25044_half_map_1.map | ||||||||||||
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Annotation | Half map A | ||||||||||||
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Density Histograms |
-Half map: Half map B
File | emd_25044_half_map_2.map | ||||||||||||
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Annotation | Half map B | ||||||||||||
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Density Histograms |
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Sample components
+Entire : Ternary complex of SHOC2:PP1C:MRAS
+Supramolecule #1: Ternary complex of SHOC2:PP1C:MRAS
+Supramolecule #2: SHOC2
+Supramolecule #3: MRAS
+Supramolecule #4: PP1C
+Macromolecule #1: Leucine-rich repeat protein SHOC-2
+Macromolecule #2: Ras-related protein M-Ras
+Macromolecule #3: Serine/threonine-protein phosphatase PP1-gamma catalytic subunit
+Macromolecule #4: PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER
+Macromolecule #5: MAGNESIUM ION
+Macromolecule #6: MANGANESE (II) ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.19 mg/mL | |||||||||||||||||||||
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Buffer | pH: 7.5 Component:
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Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Support film - Material: GRAPHENE OXIDE / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 20 sec. | |||||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV Details: Used the "perpetually hydrated" method of applying graphene oxide. (Cheung et al., 2018). |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 1 / Average electron dose: 64.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 1.5 µm / Nominal defocus min: 0.5 µm / Nominal magnification: 105000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |