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- EMDB-4737: Structure of XBP1u-paused ribosome nascent chain complex (rotated... -
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Basic information
Entry | Database: EMDB / ID: EMD-4737 | |||||||||
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Title | Structure of XBP1u-paused ribosome nascent chain complex (rotated state) | |||||||||
![]() | Structure of XBP1u-paused ribosome nascent chain complex (rotated state) | |||||||||
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![]() | translational pausing / XBP1 / UPR / RIBOSOME | |||||||||
Function / homology | ![]() epithelial cell maturation involved in salivary gland development / positive regulation of vascular wound healing / ATF6-mediated unfolded protein response / positive regulation of protein acetylation / response to insulin-like growth factor stimulus / glandular epithelial cell maturation / sterol homeostasis / positive regulation of lactation / IRE1alpha activates chaperones / ATF6 (ATF6-alpha) activates chaperone genes ...epithelial cell maturation involved in salivary gland development / positive regulation of vascular wound healing / ATF6-mediated unfolded protein response / positive regulation of protein acetylation / response to insulin-like growth factor stimulus / glandular epithelial cell maturation / sterol homeostasis / positive regulation of lactation / IRE1alpha activates chaperones / ATF6 (ATF6-alpha) activates chaperone genes / positive regulation of plasma cell differentiation / positive regulation of phospholipid biosynthetic process / positive regulation of ERAD pathway / negative regulation of myotube differentiation / intracellular triglyceride homeostasis / cellular response to fructose stimulus / cellular response to laminar fluid shear stress / XBP1(S) activates chaperone genes / cellular response to nutrient / cellular response to fluid shear stress / positive regulation of MHC class II biosynthetic process / exocrine pancreas development / negative regulation of endoplasmic reticulum unfolded protein response / positive regulation of vascular associated smooth muscle cell migration / positive regulation of hepatocyte proliferation / endothelial cell proliferation / ribosomal subunit / positive regulation of T cell differentiation / cellular response to peptide hormone stimulus / positive regulation of B cell differentiation / positive regulation of immunoglobulin production / muscle organ development / IRE1-mediated unfolded protein response / negative regulation of SMAD protein signal transduction / ubiquitin ligase inhibitor activity / positive regulation of endothelial cell apoptotic process / intrinsic apoptotic signaling pathway in response to endoplasmic reticulum stress / positive regulation of signal transduction by p53 class mediator / cellular response to vascular endothelial growth factor stimulus / positive regulation of TOR signaling / positive regulation of fat cell differentiation / 90S preribosome / adipose tissue development / fatty acid homeostasis / vascular endothelial growth factor receptor signaling pathway / neuron development / phagocytic cup / negative regulation of endoplasmic reticulum stress-induced intrinsic apoptotic signaling pathway / endoplasmic reticulum unfolded protein response / cellular response to glucose starvation / cis-regulatory region sequence-specific DNA binding / ribosomal small subunit export from nucleus / positive regulation of vascular associated smooth muscle cell proliferation / rough endoplasmic reticulum / translation regulator activity / ERAD pathway / gastrulation / MDM2/MDM4 family protein binding / positive regulation of autophagy / cytosolic ribosome / class I DNA-(apurinic or apyrimidinic site) endonuclease activity / DNA-(apurinic or apyrimidinic site) lyase / cellular response to interleukin-4 / response to endoplasmic reticulum stress / cholesterol homeostasis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / cellular response to leukemia inhibitory factor / ribosomal large subunit biogenesis / positive regulation of apoptotic signaling pathway / maturation of SSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / nuclear estrogen receptor binding / maturation of SSU-rRNA / RNA polymerase II transcription regulatory region sequence-specific DNA binding / cellular response to amino acid stimulus / regulation of cell growth / small-subunit processome / cellular response to glucose stimulus / phosphatidylinositol 3-kinase/protein kinase B signal transduction / negative regulation of transforming growth factor beta receptor signaling pathway / chromatin DNA binding / liver development / protein destabilization / negative regulation of ERK1 and ERK2 cascade / regulation of protein stability / autophagy / positive regulation of interleukin-6 production / positive regulation of protein import into nucleus / spindle / RNA polymerase II transcription regulator complex / positive regulation of protein phosphorylation / cellular response to insulin stimulus / positive regulation of angiogenesis / fatty acid biosynthetic process / rRNA processing / sequence-specific double-stranded DNA binding / rhythmic process / positive regulation of canonical Wnt signaling pathway / protein transport / ribosome biogenesis / large ribosomal subunit Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.1 Å | |||||||||
![]() | Shanmuganathan V / Cheng J | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structural and mutational analysis of the ribosome-arresting human XBP1u. Authors: Vivekanandan Shanmuganathan / Nina Schiller / Anastasia Magoulopoulou / Jingdong Cheng / Katharina Braunger / Florian Cymer / Otto Berninghausen / Birgitta Beatrix / Kenji Kohno / Gunnar von ...Authors: Vivekanandan Shanmuganathan / Nina Schiller / Anastasia Magoulopoulou / Jingdong Cheng / Katharina Braunger / Florian Cymer / Otto Berninghausen / Birgitta Beatrix / Kenji Kohno / Gunnar von Heijne / Roland Beckmann / ![]() ![]() ![]() Abstract: XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM ...XBP1u, a central component of the unfolded protein response (UPR), is a mammalian protein containing a functionally critical translational arrest peptide (AP). Here, we present a 3 Å cryo-EM structure of the stalled human XBP1u AP. It forms a unique turn in the ribosomal exit tunnel proximal to the peptidyl transferase center where it causes a subtle distortion, thereby explaining the temporary translational arrest induced by XBP1u. During ribosomal pausing the hydrophobic region 2 (HR2) of XBP1u is recognized by SRP, but fails to efficiently gate the Sec61 translocon. An exhaustive mutagenesis scan of the XBP1u AP revealed that only 8 out of 20 mutagenized positions are optimal; in the remaining 12 positions, we identify 55 different mutations increase the level of translational arrest. Thus, the wildtype XBP1u AP induces only an intermediate level of translational arrest, allowing efficient targeting by SRP without activating the Sec61 channel. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 24.4 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 109.8 KB 109.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 14.1 KB | Display | ![]() |
Images | ![]() | 232.3 KB | ||
Filedesc metadata | ![]() | 19.4 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 285.9 KB | Display | ![]() |
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Full document | ![]() | 285 KB | Display | |
Data in XML | ![]() | 13.9 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6r6pMC ![]() 4729C ![]() 4735C ![]() 4745C ![]() 6r5qC ![]() 6r6gC ![]() 6r7qC 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
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Structure of XBP1u-paused ribosome nascent chain complex (rotated state) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.084 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 : Structure of XBP1u-paused ribosome nascent chain complex (rotated...
+Supramolecule #1: Structure of XBP1u-paused ribosome nascent chain complex (rotated...
+Supramolecule #2: Structure of XBP1u-paused ribosome nascent chain complex (rotated...
+Supramolecule #3: A/P- and P/E- site tRNA
+Supramolecule #4: X-box-binding protein 1
+Supramolecule #5: mRNA
+Macromolecule #1: 28S ribosomal RNA
+Macromolecule #2: 5S ribosomal RNA
+Macromolecule #3: 5.8S ribosomal RNA
+Macromolecule #49: A/P-tRNA
+Macromolecule #50: P/E-tRNA
+Macromolecule #51: 18S rRNA
+Macromolecule #85: mRNA
+Macromolecule #4: uL2
+Macromolecule #5: uL3
+Macromolecule #6: uL4
+Macromolecule #7: 60S ribosomal protein L5
+Macromolecule #8: 60S ribosomal protein L6
+Macromolecule #9: uL30
+Macromolecule #10: eL8
+Macromolecule #11: uL6
+Macromolecule #12: Ribosomal protein L10 (Predicted)
+Macromolecule #13: uL5
+Macromolecule #14: 60S ribosomal protein L13
+Macromolecule #15: Ribosomal protein L14
+Macromolecule #16: Ribosomal protein L15
+Macromolecule #17: uL13
+Macromolecule #18: uL22
+Macromolecule #19: eL18
+Macromolecule #20: eL19
+Macromolecule #21: eL20
+Macromolecule #22: eL21
+Macromolecule #23: Ribosomal protein L22
+Macromolecule #24: uL14
+Macromolecule #25: Ribosomal protein L24
+Macromolecule #26: uL23
+Macromolecule #27: Ribosomal protein L26
+Macromolecule #28: 60S ribosomal protein L27
+Macromolecule #29: uL15
+Macromolecule #30: 60S ribosomal protein L29
+Macromolecule #31: eL30
+Macromolecule #32: eL31
+Macromolecule #33: eL32
+Macromolecule #34: eL33
+Macromolecule #35: eL34
+Macromolecule #36: uL29
+Macromolecule #37: 60S ribosomal protein L36
+Macromolecule #38: Ribosomal protein L37
+Macromolecule #39: eL38
+Macromolecule #40: ribosomal protein eL39
+Macromolecule #41: eL40
+Macromolecule #42: 60s ribosomal protein l41
+Macromolecule #43: eL42
+Macromolecule #44: ribosomal protein eL43
+Macromolecule #45: eL28
+Macromolecule #46: 60S acidic ribosomal protein P0
+Macromolecule #47: Ribosomal protein L12
+Macromolecule #48: X-box-binding protein 1
+Macromolecule #52: uS2
+Macromolecule #53: 40S ribosomal protein S3a
+Macromolecule #54: uS5
+Macromolecule #55: 40S ribosomal protein S4
+Macromolecule #56: 40S ribosomal protein S6
+Macromolecule #57: 40S ribosomal protein S7
+Macromolecule #58: 40S ribosomal protein S8
+Macromolecule #59: Ribosomal protein S9 (Predicted)
+Macromolecule #60: Ribosomal protein S11
+Macromolecule #61: ribosomal protein uS15
+Macromolecule #62: uS11
+Macromolecule #63: Ribosomal protein S16
+Macromolecule #64: eS17
+Macromolecule #65: eS21
+Macromolecule #66: Ribosomal protein S15a
+Macromolecule #67: Ribosomal protein S23
+Macromolecule #68: eS24
+Macromolecule #69: eS26
+Macromolecule #70: 40S ribosomal protein S27
+Macromolecule #71: 40S ribosomal protein S30
+Macromolecule #72: Ribosomal protein S3
+Macromolecule #73: Ribosomal protein S5
+Macromolecule #74: eS10
+Macromolecule #75: 40S ribosomal protein S12
+Macromolecule #76: uS14
+Macromolecule #77: uS13
+Macromolecule #78: eS19
+Macromolecule #79: uS10
+Macromolecule #80: ribosomal protein eS25
+Macromolecule #81: Ribosomal protein S28
+Macromolecule #82: S29
+Macromolecule #83: eS31
+Macromolecule #84: ribosomal protein RACK1
+Macromolecule #86: MAGNESIUM ION
+Macromolecule #87: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Average electron dose: 28.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Protocol: RIGID BODY FIT |
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Output model | ![]() PDB-6r6p: |