+Open data
-Basic information
Entry | Database: PDB / ID: 6lt0 | ||||||||||||
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Title | cryo-EM structure of C9ORF72-SMCR8-WDR41 | ||||||||||||
Components |
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Keywords | PROTEIN BINDING / ALS / FTD / GTPase / C9ORF72 / SMCR8 / WDR41 / GAP / GEF | ||||||||||||
Function / homology | Function and homology information Atg1/ULK1 kinase complex / late endosome to lysosome transport / regulation of TORC1 signaling / regulation of autophagosome assembly / negative regulation of autophagosome assembly / regulation of actin filament organization / guanyl-nucleotide exchange factor complex / negative regulation of immune response / axon extension / negative regulation of exocytosis ...Atg1/ULK1 kinase complex / late endosome to lysosome transport / regulation of TORC1 signaling / regulation of autophagosome assembly / negative regulation of autophagosome assembly / regulation of actin filament organization / guanyl-nucleotide exchange factor complex / negative regulation of immune response / axon extension / negative regulation of exocytosis / Flemming body / positive regulation of autophagosome maturation / main axon / negative regulation of macroautophagy / protein kinase inhibitor activity / positive regulation of macroautophagy / positive regulation of TOR signaling / autophagosome / stress granule assembly / axonal growth cone / GTPase activator activity / guanyl-nucleotide exchange factor activity / negative regulation of protein phosphorylation / regulation of autophagy / cell projection / P-body / negative regulation of protein kinase activity / small GTPase binding / autophagy / positive regulation of GTPase activity / cytoplasmic stress granule / endocytosis / regulation of protein localization / presynapse / perikaryon / postsynapse / nuclear membrane / lysosome / endosome / lysosomal membrane / negative regulation of gene expression / intracellular membrane-bounded organelle / dendrite / chromatin / protein kinase binding / extracellular space / nucleoplasm / nucleus / cytosol / cytoplasm Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) | ||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.2 Å | ||||||||||||
Authors | Tang, D. / Sheng, J. / Xu, L. / Zhan, X. / Yan, C. / Qi, S. | ||||||||||||
Funding support | China, 3items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2020 Title: Cryo-EM structure of C9ORF72-SMCR8-WDR41 reveals the role as a GAP for Rab8a and Rab11a. Authors: Dan Tang / Jingwen Sheng / Liangting Xu / Xiechao Zhan / Jiaming Liu / Hui Jiang / Xiaoling Shu / Xiaoyu Liu / Tizhong Zhang / Lan Jiang / Cuiyan Zhou / Wenqi Li / Wei Cheng / Zhonghan Li / ...Authors: Dan Tang / Jingwen Sheng / Liangting Xu / Xiechao Zhan / Jiaming Liu / Hui Jiang / Xiaoling Shu / Xiaoyu Liu / Tizhong Zhang / Lan Jiang / Cuiyan Zhou / Wenqi Li / Wei Cheng / Zhonghan Li / Kunjie Wang / Kefeng Lu / Chuangye Yan / Shiqian Qi / Abstract: A massive intronic hexanucleotide repeat (GGGGCC) expansion in is a genetic origin of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recently, C9ORF72, together with SMCR8 ...A massive intronic hexanucleotide repeat (GGGGCC) expansion in is a genetic origin of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recently, C9ORF72, together with SMCR8 and WDR41, has been shown to regulate autophagy and function as Rab GEF. However, the precise function of C9ORF72 remains unclear. Here, we report the cryogenic electron microscopy (cryo-EM) structure of the human C9ORF72-SMCR8-WDR41 complex at a resolution of 3.2 Å. The structure reveals the dimeric assembly of a heterotrimer of C9ORF72-SMCR8-WDR41. Notably, the C-terminal tail of C9ORF72 and the DENN domain of SMCR8 play critical roles in the dimerization of the two protomers of the C9ORF72-SMCR8-WDR41 complex. In the protomer, C9ORF72 and WDR41 are joined by SMCR8 without direct interaction. WDR41 binds to the DENN domain of SMCR8 by the C-terminal helix. Interestingly, the prominent structural feature of C9ORF72-SMCR8 resembles that of the FLNC-FNIP2 complex, the GTPase activating protein (GAP) of RagC/D. Structural comparison and sequence alignment revealed that Arg147 of SMCR8 is conserved and corresponds to the arginine finger of FLCN, and biochemical analysis indicated that the Arg147 of SMCR8 is critical to the stimulatory effect of the C9ORF72-SMCR8 complex on Rab8a and Rab11a. Our study not only illustrates the basis of C9ORF72-SMCR8-WDR41 complex assembly but also reveals the GAP activity of the C9ORF72-SMCR8 complex. | ||||||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
-Download
PDBx/mmCIF format | 6lt0.cif.gz | 420.5 KB | Display | PDBx/mmCIF format |
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PDB format | pdb6lt0.ent.gz | 326.5 KB | Display | PDB format |
PDBx/mmJSON format | 6lt0.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/lt/6lt0 ftp://data.pdbj.org/pub/pdb/validation_reports/lt/6lt0 | HTTPS FTP |
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-Related structure data
Related structure data | 0966MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 51783.805 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: WDR41, MSTP048 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q9HAD4 #2: Protein | Mass: 105149.094 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SMCR8 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q8TEV9 #3: Protein | Mass: 54391.477 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: C9orf72 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q96LT7 |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Dimer of C9-ORF72-SMCR8-WDR41 / Type: COMPLEX / Entity ID: all / Source: RECOMBINANT | ||||||||||||||||
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Molecular weight | Value: 0.4 MDa / Experimental value: YES | ||||||||||||||||
Source (natural) | Organism: Homo sapiens (human) | ||||||||||||||||
Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) | ||||||||||||||||
Buffer solution | pH: 8 | ||||||||||||||||
Buffer component |
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Specimen | Conc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: monodisperse | ||||||||||||||||
Specimen support | Grid material: GOLD / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | ||||||||||||||||
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: SPOT SCAN |
Electron lens | Mode: DIFFRACTION |
Specimen holder | Cryogen: HELIUM / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 0.25 sec. / Electron dose: 5.3 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 5332 |
Image scans | Movie frames/image: 32 |
-Processing
Software | Name: PHENIX / Version: 1.11.1_2575: / Classification: refinement | ||||||||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.2 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 347925 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: RIGID BODY FIT / Space: REAL Details: The atomic coordinates of the CSW complex was generated by combining homology modelling and de novo model building. | ||||||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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