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Yorodumi- PDB-8i7r: In situ structure of axonemal doublet microtubules in mouse sperm... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 8i7r | |||||||||||||||||||||
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| Title | In situ structure of axonemal doublet microtubules in mouse sperm with 48-nm repeat | |||||||||||||||||||||
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Keywords | STRUCTURAL PROTEIN / microtubules / axoneme / sperm / filament | |||||||||||||||||||||
| Function / homology | Function and homology informationprotein localization to motile cilium / left/right pattern formation / axonemal microtubule doublet inner sheath / outer acrosomal membrane / epithelial cilium movement involved in determination of left/right asymmetry / regulation of brood size / establishment of left/right asymmetry / 9+0 motile cilium / sperm flagellum assembly / manchette assembly ...protein localization to motile cilium / left/right pattern formation / axonemal microtubule doublet inner sheath / outer acrosomal membrane / epithelial cilium movement involved in determination of left/right asymmetry / regulation of brood size / establishment of left/right asymmetry / 9+0 motile cilium / sperm flagellum assembly / manchette assembly / axonemal B tubule inner sheath / axonemal A tubule inner sheath / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / protein polyglutamylation / regulation of calcineurin-NFAT signaling cascade / sperm axoneme assembly / Carboxyterminal post-translational modifications of tubulin / regulation of microtubule nucleation / Intraflagellar transport / positive regulation of feeding behavior / COPI-independent Golgi-to-ER retrograde traffic / MAP kinase tyrosine/serine/threonine phosphatase activity / inner dynein arm assembly / cilium-dependent cell motility / cerebrospinal fluid circulation / regulation of cilium beat frequency involved in ciliary motility / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / epithelial cilium movement involved in extracellular fluid movement / 9+2 motile cilium / COPI-mediated anterograde transport / cilium movement involved in cell motility / Aggrephagy / intraciliary transport / regulation of store-operated calcium entry / Kinesins / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Resolution of Sister Chromatid Cohesion / PKR-mediated signaling / acrosomal membrane / axoneme assembly / left/right axis specification / cilium movement / The role of GTSE1 in G2/M progression after G2 checkpoint / RHO GTPases activate IQGAPs / microtubule sliding / Recycling pathway of L1 / calcium ion sensor activity / COPI-dependent Golgi-to-ER retrograde traffic / RHO GTPases Activate Formins / Separation of Sister Chromatids / axonemal microtubule / Hedgehog 'off' state / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Anchoring of the basal body to the plasma membrane / Recruitment of NuMA to mitotic centrosomes / cilium organization / AURKA Activation by TPX2 / gamma-tubulin ring complex / Regulation of PLK1 Activity at G2/M Transition / manchette / positive regulation of cilium assembly / MHC class II antigen presentation / flagellated sperm motility / 3'-5'-DNA exonuclease activity / UTP biosynthetic process / CTP biosynthetic process / motile cilium / determination of left/right symmetry / intermediate filament / DNA catabolic process / protein targeting to membrane / extrinsic component of membrane / nucleoside diphosphate kinase activity / positive regulation of cell motility / : / GTP biosynthetic process / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / tubulin complex / protein-serine/threonine phosphatase / AMP binding / ciliary base / regulation of neuron projection development / receptor clustering / ciliary transition zone / protein serine/threonine phosphatase activity / cerebral cortex cell migration / phosphatase activity / mitotic cytokinesis / microtubule organizing center / cellular response to UV-C / regulation of cell division / cilium assembly / cellular response to unfolded protein / spermatid development / negative regulation of protein binding Similarity search - Function | |||||||||||||||||||||
| Biological species | ![]() | |||||||||||||||||||||
| Method | ELECTRON MICROSCOPY / subtomogram averaging / cryo EM / Resolution: 6.5 Å | |||||||||||||||||||||
Authors | Zhu, Y. / Yin, G.L. / Tai, L.H. / Sun, F. | |||||||||||||||||||||
| Funding support | China, 1items
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Citation | Journal: Cell Discov / Year: 2023Title: In-cell structural insight into the stability of sperm microtubule doublet. Authors: Linhua Tai / Guoliang Yin / Xiaojun Huang / Fei Sun / Yun Zhu / ![]() Abstract: The propulsion for mammalian sperm swimming is generated by flagella beating. Microtubule doublets (DMTs) along with microtubule inner proteins (MIPs) are essential structural blocks of flagella. ...The propulsion for mammalian sperm swimming is generated by flagella beating. Microtubule doublets (DMTs) along with microtubule inner proteins (MIPs) are essential structural blocks of flagella. However, the intricate molecular architecture of intact sperm DMT remains elusive. Here, by in situ cryo-electron tomography, we solved the in-cell structure of mouse sperm DMT at 4.5-7.5 Å resolutions, and built its model with 36 kinds of MIPs in 48 nm periodicity. We identified multiple copies of Tektin5 that reinforce Tektin bundle, and multiple MIPs with different periodicities that anchor the Tektin bundle to tubulin wall. This architecture contributes to a superior stability of A-tubule than B-tubule of DMT, which was revealed by structural comparison of DMTs from the intact and deformed axonemes. Our work provides an overall molecular picture of intact sperm DMT in 48 nm periodicity that is essential to understand the molecular mechanism of sperm motility as well as the related ciliopathies. | |||||||||||||||||||||
| History |
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 8i7r.cif.gz | 27.1 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb8i7r.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 8i7r.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/i7/8i7r ftp://data.pdbj.org/pub/pdb/validation_reports/i7/8i7r | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 35230MC ![]() 8i7oC C: citing same article ( M: map data used to model this data |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
+Protein , 24 types, 416 molecules ABA1A2A3A4ABADAFAHAJALBBBDBFBHBJBLCBCDCFCHCJCLDBDDDFDHDJDL...
-EF-hand domain-containing family member ... , 2 types, 5 molecules EFG4G5G6
| #9: Protein | Mass: 95891.961 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #14: Protein | Mass: 87758.023 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) ![]() |
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-Cilia- and flagella-associated protein ... , 10 types, 27 molecules GHIJKLN1N2N3N4P1P2P3UVWXXAXBXCXDXEXFXGYZa
| #12: Protein | Mass: 62036.609 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #16: Protein | | Mass: 26633.035 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() #18: Protein | | Mass: 23062.510 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() #20: Protein | Mass: 34433.383 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #26: Protein | Mass: 18960.092 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Source: (natural) ![]() #29: Protein | Mass: 68322.164 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Source: (natural) ![]() #33: Protein | Mass: 65962.016 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Source: (natural) ![]() #34: Protein | Mass: 22781.389 Da / Num. of mol.: 7 / Source method: isolated from a natural source / Source: (natural) ![]() #36: Protein | Mass: 65266.520 Da / Num. of mol.: 2 / Source method: isolated from a natural source / Source: (natural) ![]() #38: Protein | | Mass: 12278.145 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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-Piercer of microtubule wall ... , 2 types, 2 molecules MN
| #23: Protein | Mass: 18862.852 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
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| #25: Protein | Mass: 13728.513 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) ![]() |
-Non-polymers , 1 types, 279 molecules 
| #39: Chemical | ChemComp-GTP / |
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-Details
| Has ligand of interest | N |
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| Has protein modification | Y |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: CELL / 3D reconstruction method: subtomogram averaging |
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Sample preparation
| Component | Name: mouse sperm / Type: CELL / Entity ID: #1-#38 / Source: NATURAL |
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| Source (natural) | Organism: ![]() |
| Buffer solution | pH: 7 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | 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: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: SPOT SCAN |
| Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 5000 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm |
| Image recording | Electron dose: 3 e/Å2 / Avg electron dose per subtomogram: 117 e/Å2 / Detector mode: SUPER-RESOLUTION / Film or detector model: GATAN K2 QUANTUM (4k x 4k) |
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Processing
| Software | Name: UCSF ChimeraX / Version: 1.6/v9 / Classification: model building / URL: https://www.rbvi.ucsf.edu/chimerax/ / Os: Windows / Type: package |
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
| Symmetry | Point symmetry: C1 (asymmetric) |
| 3D reconstruction | Resolution: 6.5 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 17450 / Symmetry type: POINT |
| EM volume selection | Num. of tomograms: 689 / Num. of volumes extracted: 17450 |
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FIELD EMISSION GUN