+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-29013 | |||||||||
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Title | 96nm doublet microtubule repeat from wild type mouse sperm | |||||||||
Map data | 96nm doublet microtubule repeat from WT mouse sperm | |||||||||
Sample |
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Keywords | 96 nm doublet microtubule from mouse sperm / STRUCTURAL PROTEIN | |||||||||
Biological species | Mus (mice) | |||||||||
Method | subtomogram averaging / cryo EM / Resolution: 34.0 Å | |||||||||
Authors | Hwang JY / Chai P / Nawaz S | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Elife / Year: 2023 Title: LRRC23 truncation impairs radial spoke 3 head assembly and sperm motility underlying male infertility. Authors: Jae Yeon Hwang / Pengxin Chai / Shoaib Nawaz / Jungmin Choi / Francesc Lopez-Giraldez / Shabir Hussain / Kaya Bilguvar / Shrikant Mane / Richard P Lifton / Wasim Ahmad / Kai Zhang / Jean-Ju Chung / Abstract: Radial spokes (RS) are T-shaped multiprotein complexes on the axonemal microtubules. Repeated RS1, RS2, and RS3 couple the central pair to modulate ciliary and flagellar motility. Despite the cell ...Radial spokes (RS) are T-shaped multiprotein complexes on the axonemal microtubules. Repeated RS1, RS2, and RS3 couple the central pair to modulate ciliary and flagellar motility. Despite the cell type specificity of RS3 substructures, their molecular components remain largely unknown. Here, we report that a leucine-rich repeat-containing protein, LRRC23, is an RS3 head component essential for its head assembly and flagellar motility in mammalian spermatozoa. From infertile male patients with defective sperm motility, we identified a splice site variant of . A mutant mouse model mimicking this variant produces a truncated LRRC23 at the C-terminus that fails to localize to the sperm tail, causing male infertility due to defective sperm motility. LRRC23 was previously proposed to be an ortholog of the RS stalk protein RSP15. However, we found that purified recombinant LRRC23 interacts with an RS head protein RSPH9, which is abolished by the C-terminal truncation. Evolutionary and structural comparison also shows that LRRC34, not LRRC23, is the RSP15 ortholog. Cryo-electron tomography clearly revealed that the absence of the RS3 head and the sperm-specific RS2-RS3 bridge structure in LRRC23 mutant spermatozoa. Our study provides new insights into the structure and function of RS3 in mammalian spermatozoa and the molecular pathogenicity of LRRC23 underlying reduced sperm motility in infertile human males. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_29013.map.gz | 2.5 MB | EMDB map data format | |
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Header (meta data) | emd-29013-v30.xml emd-29013.xml | 16.6 KB 16.6 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_29013_fsc.xml | 5.8 KB | Display | FSC data file |
Images | emd_29013.png | 58.8 KB | ||
Filedesc metadata | emd-29013.cif.gz | 4.3 KB | ||
Others | emd_29013_additional_1.map.gz emd_29013_half_map_1.map.gz emd_29013_half_map_2.map.gz | 14.7 MB 7.7 MB 7.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-29013 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29013 | HTTPS FTP |
-Validation report
Summary document | emd_29013_validation.pdf.gz | 665.9 KB | Display | EMDB validaton report |
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Full document | emd_29013_full_validation.pdf.gz | 665.4 KB | Display | |
Data in XML | emd_29013_validation.xml.gz | 11.6 KB | Display | |
Data in CIF | emd_29013_validation.cif.gz | 15.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29013 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29013 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_29013.map.gz / Format: CCP4 / Size: 15.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | 96nm doublet microtubule repeat from WT mouse sperm | ||||||||||||||||||||
Voxel size | X=Y=Z: 10.2 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: unmasked 96nm doublet microtubule repeat from WT mouse sperm
File | emd_29013_additional_1.map | ||||||||||||
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Annotation | unmasked 96nm doublet microtubule repeat from WT mouse sperm | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map 2 of 96nm doublet microtubule repeat from WT mouse sperm
File | emd_29013_half_map_1.map | ||||||||||||
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Annotation | half map 2 of 96nm doublet microtubule repeat from WT mouse sperm | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map 1 of 96nm doublet microtubule repeat from WT mouse sperm
File | emd_29013_half_map_2.map | ||||||||||||
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Annotation | half map 1 of 96nm doublet microtubule repeat from WT mouse sperm | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Mouse sperm
Entire | Name: Mouse sperm |
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Components |
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-Supramolecule #1: Mouse sperm
Supramolecule | Name: Mouse sperm / type: cell / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Mus (mice) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 5.5 |
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Grid | Model: Quantifoil R2/2 / Material: GOLD / Mesh: 300 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 30 % / Chamber temperature: 298 K / Instrument: HOMEMADE PLUNGER |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 3.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 0.5 µm / Nominal defocus min: 0.2 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |