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- EMDB-35230: In situ structure of axonemal doublet microtubules in mouse sperm... -
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Basic information
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Title | In situ structure of axonemal doublet microtubules in mouse sperm with 48-nm repeat | |||||||||
![]() | In situ structure of axonemal doublet microtubules in mouse sperm with 48-nm repeat | |||||||||
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![]() | microtubules / axoneme / sperm / filament / STRUCTURAL PROTEIN | |||||||||
Function / homology | ![]() male germ-line stem cell population maintenance / left/right pattern formation / axonemal microtubule doublet inner sheath / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / outer acrosomal membrane / epithelial cilium movement involved in determination of left/right asymmetry / sperm flagellum assembly / regulation of brood size ...male germ-line stem cell population maintenance / left/right pattern formation / axonemal microtubule doublet inner sheath / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / outer acrosomal membrane / epithelial cilium movement involved in determination of left/right asymmetry / sperm flagellum assembly / regulation of brood size / establishment of left/right asymmetry / 9+0 motile cilium / protein localization to motile cilium / manchette assembly / axonemal B tubule inner sheath / Intraflagellar transport / axonemal A tubule inner sheath / manchette / Carboxyterminal post-translational modifications of tubulin / sperm axoneme assembly / protein polyglutamylation / inner dynein arm assembly / positive regulation of feeding behavior / regulation of calcineurin-NFAT signaling cascade / COPI-independent Golgi-to-ER retrograde traffic / regulation of microtubule nucleation / sperm principal piece / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / MAP kinase tyrosine/serine/threonine phosphatase activity / microtubule sliding / cerebrospinal fluid circulation / regulation of cilium beat frequency involved in ciliary motility / epithelial cilium movement involved in extracellular fluid movement / cilium movement involved in cell motility / COPI-mediated anterograde transport / Aggrephagy / protein localization to organelle / regulation of store-operated calcium entry / Kinesins / Transferases; Transferring phosphorus-containing groups / 9+2 motile cilium / intraciliary transport / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / PKR-mediated signaling / acrosomal membrane / Resolution of Sister Chromatid Cohesion / The role of GTSE1 in G2/M progression after G2 checkpoint / cilium movement / ciliary transition zone / RHO GTPases activate IQGAPs / Recycling pathway of L1 / axoneme assembly / left/right axis specification / COPI-dependent Golgi-to-ER retrograde traffic / cilium organization / axonemal microtubule / calcium ion sensor activity / RHO GTPases Activate Formins / Separation of Sister Chromatids / Hedgehog 'off' state / gamma-tubulin ring complex / Loss of Nlp from mitotic centrosomes / Recruitment of mitotic centrosome proteins and complexes / Loss of proteins required for interphase microtubule organization from the centrosome / Recruitment of NuMA to mitotic centrosomes / Anchoring of the basal body to the plasma membrane / AURKA Activation by TPX2 / Regulation of PLK1 Activity at G2/M Transition / flagellated sperm motility / positive regulation of cilium assembly / cell projection organization / MHC class II antigen presentation / UTP biosynthetic process / CTP biosynthetic process / determination of left/right symmetry / protein targeting to membrane / protein targeting to mitochondrion / intermediate filament / extrinsic component of membrane / nucleoside diphosphate kinase activity / GTP biosynthetic process / positive regulation of cell motility / tubulin complex / histone H2AXS140 phosphatase activity / RNA polymerase II CTD heptapeptide repeat Y1 phosphatase activity / RNA polymerase II CTD heptapeptide repeat T4 phosphatase activity / RNA polymerase II CTD heptapeptide repeat S2 phosphatase activity / RNA polymerase II CTD heptapeptide repeat S5 phosphatase activity / RNA polymerase II CTD heptapeptide repeat S7 phosphatase activity / MAP kinase serine/threonine phosphatase activity / calmodulin-dependent protein phosphatase activity / myosin phosphatase activity / protein-serine/threonine phosphatase / AMP binding / ciliary base / receptor clustering / intercellular bridge / beta-tubulin binding / spermatid development Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 6.5 Å | |||||||||
![]() | Zhu Y / Yin GL / Tai LH / Sun F | |||||||||
Funding support | ![]()
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![]() | ![]() Title: 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
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 56.3 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 57.4 KB 57.4 KB | Display Display | ![]() |
Images | ![]() | 165.9 KB | ||
Filedesc metadata | ![]() | 16.6 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 636.6 KB | Display | ![]() |
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Full document | ![]() | 636.2 KB | Display | |
Data in XML | ![]() | 6.5 KB | Display | |
Data in CIF | ![]() | 7.4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8i7rMC ![]() 8i7oC C: citing same article ( M: atomic model generated by this map |
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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 | In situ structure of axonemal doublet microtubules in mouse sperm with 48-nm repeat | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.76 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
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Sample components
+Entire : mouse sperm
+Supramolecule #1: mouse sperm
+Macromolecule #1: Meiosis-specific nuclear structural protein 1
+Macromolecule #2: Tektin-1
+Macromolecule #3: Tubulin beta-4B chain
+Macromolecule #4: Detyrosinated tubulin alpha-3 chain
+Macromolecule #5: Tektin-2
+Macromolecule #6: Nucleoside diphosphate kinase 7
+Macromolecule #7: Tektin-3
+Macromolecule #8: Tektin-4
+Macromolecule #9: EF-hand domain-containing family member B
+Macromolecule #10: Tektin bundle-interacting protein 1
+Macromolecule #11: Tektin-5
+Macromolecule #12: Cilia- and flagella-associated protein 53
+Macromolecule #13: EF-hand domain-containing protein 1
+Macromolecule #14: EF-hand domain-containing family member C2
+Macromolecule #15: Protein FAM166A
+Macromolecule #16: Cilia- and flagella-associated protein 95
+Macromolecule #17: Protein FAM166C
+Macromolecule #18: Cilia- and flagella-associated protein 107
+Macromolecule #19: Dual specificity phosphatase 21
+Macromolecule #20: Cilia- and flagella-associated protein 161
+Macromolecule #21: Coiled-coil domain-containing protein 105
+Macromolecule #22: Enkurin
+Macromolecule #23: Piercer of microtubule wall 1 protein
+Macromolecule #24: Testis-expressed protein 43
+Macromolecule #25: Piercer of microtubule wall 2 protein
+Macromolecule #26: Cilia- and flagella-associated protein 276
+Macromolecule #27: RIB43A-like with coiled-coils protein 2
+Macromolecule #28: Protein Flattop
+Macromolecule #29: Cilia- and flagella-associated protein 52
+Macromolecule #30: EF-hand calcium-binding domain-containing protein 6
+Macromolecule #31: Cilia and flagella-associated protein 77
+Macromolecule #32: Sperm-associated antigen 8
+Macromolecule #33: Cilia- and flagella-associated protein 45
+Macromolecule #34: Cilia- and flagella-associated protein 20
+Macromolecule #35: Parkin coregulated gene protein homolog
+Macromolecule #36: Cilia- and flagella-associated protein 210
+Macromolecule #37: Sperm acrosome-associated protein 9
+Macromolecule #38: Cilia- and flagella-associated protein 141
+Macromolecule #39: GUANOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | subtomogram averaging |
Aggregation state | cell |
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Sample preparation
Buffer | pH: 7 |
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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: GATAN K2 QUANTUM (4k x 4k) / Detector mode: SUPER-RESOLUTION / Average electron dose: 3.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 5.0 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 6.5 Å / Resolution method: FSC 0.143 CUT-OFF / Number subtomograms used: 17450 |
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Extraction | Number tomograms: 689 / Number images used: 17450 |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |