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- PDB-8oj5: 60S ribosomal subunit bound to the E3-UFM1 complex - state 3 (in-... -
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Open data
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Basic information
Entry | Database: PDB / ID: 8oj5 | ||||||||||||||||||
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Title | 60S ribosomal subunit bound to the E3-UFM1 complex - state 3 (in-vitro reconstitution) | ||||||||||||||||||
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![]() | RIBOSOME / ER / UFMylation / recycling | ||||||||||||||||||
Function / homology | ![]() UFM1 ligase activity / UFM1-modified protein reader activity / positive regulation of reticulophagy / regulation of phosphatase activity / apoptotic nuclear changes / UFM1 transferase activity / definitive erythrocyte differentiation / positive regulation of metallopeptidase activity / protein K69-linked ufmylation / protein ufmylation ...UFM1 ligase activity / UFM1-modified protein reader activity / positive regulation of reticulophagy / regulation of phosphatase activity / apoptotic nuclear changes / UFM1 transferase activity / definitive erythrocyte differentiation / positive regulation of metallopeptidase activity / protein K69-linked ufmylation / protein ufmylation / : / positive regulation of protein localization to endoplasmic reticulum / positive regulation of proteolysis involved in protein catabolic process / negative regulation of protein kinase activity by regulation of protein phosphorylation / positive regulation of I-kappaB phosphorylation / positive regulation of plasma cell differentiation / negative regulation of IRE1-mediated unfolded protein response / regulation of proteasomal ubiquitin-dependent protein catabolic process / negative regulation of T cell mediated immune response to tumor cell / protein localization to endoplasmic reticulum / positive regulation of cell cycle G1/S phase transition / negative regulation of T cell activation / regulation of intracellular estrogen receptor signaling pathway / negative regulation of protein serine/threonine kinase activity / eukaryotic 80S initiation complex / negative regulation of protein neddylation / positive regulation of proteasomal protein catabolic process / negative regulation of formation of translation preinitiation complex / regulation of G1 to G0 transition / axial mesoderm development / ribosomal protein import into nucleus / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / regulation of translation involved in cellular response to UV / mitotic G2/M transition checkpoint / protein-DNA complex disassembly / 90S preribosome assembly / : / Transferases; Acyltransferases; Aminoacyltransferases / ribosome disassembly / TORC2 complex binding / GAIT complex / A band / regulation of canonical NF-kappaB signal transduction / cartilage development / mitogen-activated protein kinase binding / reticulophagy / G1 to G0 transition / middle ear morphogenesis / alpha-beta T cell differentiation / regulation of neuron differentiation / translation at presynapse / exit from mitosis / cytoplasmic side of rough endoplasmic reticulum membrane / optic nerve development / negative regulation of protein import into nucleus / response to L-glutamate / negative regulation of ubiquitin protein ligase activity / retinal ganglion cell axon guidance / regulation of cyclin-dependent protein serine/threonine kinase activity / response to aldosterone / homeostatic process / macrophage chemotaxis / negative regulation of NF-kappaB transcription factor activity / lung morphogenesis / mitotic G2 DNA damage checkpoint signaling / negative regulation of PERK-mediated unfolded protein response / male meiosis I / Protein hydroxylation / ubiquitin-like protein ligase binding / Peptide chain elongation / Selenocysteine synthesis / positive regulation of signal transduction by p53 class mediator / Formation of a pool of free 40S subunits / ubiquitin ligase inhibitor activity / Eukaryotic Translation Termination / blastocyst development / Response of EIF2AK4 (GCN2) to amino acid deficiency / SRP-dependent cotranslational protein targeting to membrane / cellular response to actinomycin D / negative regulation of ubiquitin-dependent protein catabolic process / Viral mRNA Translation / hematopoietic stem cell differentiation / RHOA GTPase cycle / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / protein localization to nucleus / GTP hydrolysis and joining of the 60S ribosomal subunit / negative regulation of MAP kinase activity / L13a-mediated translational silencing of Ceruloplasmin expression / NF-kappaB binding / Major pathway of rRNA processing in the nucleolus and cytosol / ubiquitin-like ligase-substrate adaptor activity / protein targeting / protein-RNA complex assembly / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / negative regulation of protein phosphorylation / maturation of LSU-rRNA / : / endoplasmic reticulum unfolded protein response / rough endoplasmic reticulum / positive regulation of glial cell proliferation Similarity search - Function | ||||||||||||||||||
Biological species | ![]() | ||||||||||||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.9 Å | ||||||||||||||||||
![]() | Penchev, I. / DaRosa, P.A. / Peter, J.J. / Kulathu, Y. / Becker, T. / Beckmann, R. / Kopito, R. | ||||||||||||||||||
Funding support | European Union, ![]() ![]() ![]()
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![]() | ![]() Title: UFM1 E3 ligase promotes recycling of 60S ribosomal subunits from the ER. Authors: Paul A DaRosa / Ivan Penchev / Samantha C Gumbin / Francesco Scavone / Magda Wąchalska / Joao A Paulo / Alban Ordureau / Joshua J Peter / Yogesh Kulathu / J Wade Harper / Thomas Becker / ...Authors: Paul A DaRosa / Ivan Penchev / Samantha C Gumbin / Francesco Scavone / Magda Wąchalska / Joao A Paulo / Alban Ordureau / Joshua J Peter / Yogesh Kulathu / J Wade Harper / Thomas Becker / Roland Beckmann / Ron R Kopito / ![]() ![]() ![]() Abstract: Reversible modification of target proteins by ubiquitin and ubiquitin-like proteins (UBLs) is widely used by eukaryotic cells to control protein fate and cell behaviour. UFM1 is a UBL that ...Reversible modification of target proteins by ubiquitin and ubiquitin-like proteins (UBLs) is widely used by eukaryotic cells to control protein fate and cell behaviour. UFM1 is a UBL that predominantly modifies a single lysine residue on a single ribosomal protein, uL24 (also called RPL26), on ribosomes at the cytoplasmic surface of the endoplasmic reticulum (ER). UFM1 conjugation (UFMylation) facilitates the rescue of 60S ribosomal subunits (60S) that are released after ribosome-associated quality-control-mediated splitting of ribosomes that stall during co-translational translocation of secretory proteins into the ER. Neither the molecular mechanism by which the UFMylation machinery achieves such precise target selection nor how this ribosomal modification promotes 60S rescue is known. Here we show that ribosome UFMylation in vivo occurs on free 60S and we present sequential cryo-electron microscopy snapshots of the heterotrimeric UFM1 E3 ligase (E3(UFM1)) engaging its substrate uL24. E3(UFM1) binds the L1 stalk, empty transfer RNA-binding sites and the peptidyl transferase centre through carboxy-terminal domains of UFL1, which results in uL24 modification more than 150 Å away. After catalysing UFM1 transfer, E3(UFM1) remains stably bound to its product, UFMylated 60S, forming a C-shaped clamp that extends all the way around the 60S from the transfer RNA-binding sites to the polypeptide tunnel exit. Our structural and biochemical analyses suggest a role for E3(UFM1) in post-termination release and recycling of the large ribosomal subunit from the ER membrane. | ||||||||||||||||||
History |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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PDBx/mmCIF format | ![]() | 3.6 MB | Display | ![]() |
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PDB format | ![]() | Display | ![]() | |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
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-Validation report
Summary document | ![]() | 1.8 MB | Display | ![]() |
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Full document | ![]() | 1.8 MB | Display | |
Data in XML | ![]() | 219 KB | Display | |
Data in CIF | ![]() | 383.7 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 16905MC ![]() 8ohdC ![]() 8oj0C ![]() 8oj8C C: citing same article ( M: map data used to model this data |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Components
-RNA chain , 3 types, 3 molecules 578
#1: RNA chain | Mass: 1640166.875 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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#2: RNA chain | Mass: 38998.078 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#3: RNA chain | Mass: 50449.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Protein , 6 types, 6 molecules ABCDLILm
#4: Protein | Mass: 89722.203 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() References: UniProt: O94874, Transferases; Acyltransferases; Aminoacyltransferases |
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#5: Protein | Mass: 56977.395 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() |
#6: Protein | Mass: 35663.840 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() |
#7: Protein | Mass: 9128.552 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() |
#16: Protein | Mass: 24552.910 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
#45: Protein | Mass: 14758.394 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
+60S ribosomal protein ... , 40 types, 40 molecules LALBLCLDLELFLGLHLJLLLMLNLOLPLQLRLSLTLULVLWLXLYLZLaLbLcLdLeLf...
-Non-polymers , 2 types, 225 molecules 


#50: Chemical | ChemComp-MG / #51: Chemical | ChemComp-ZN / |
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-Details
Has ligand of interest | N |
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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 |
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Sample preparation
Component | Name: E3-UFM1 complex bound to the 60S ribosome / Type: RIBOSOME / Entity ID: #1-#49 / Source: NATURAL |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: ![]() |
Buffer solution | pH: 7 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE |
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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: ![]() |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 2000 nm / Nominal defocus min: 800 nm |
Image recording | Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) |
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Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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3D reconstruction | Resolution: 2.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 35935 / Symmetry type: POINT |
Atomic model building | Protocol: AB INITIO MODEL |