- EMDB-64679: Cryo-EM structure of the 48-nm repeat doublet microtubule from mo... -
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Basic information
Entry
Database: EMDB / ID: EMD-64679
Title
Cryo-EM structure of the 48-nm repeat doublet microtubule from mouse sperm
Map data
Sample
Complex: doublet microtubule of mouse sperm flagella
Protein or peptide: x 60 types
Ligand: x 4 types
Keywords
sperm flagellar doublet microtubules / STRUCTURAL PROTEIN
Function / homology
Function and homology information
protein localization to motile cilium / 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 ...protein localization to motile cilium / 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 / outer dense fiber / Microtubule-dependent trafficking of connexons from Golgi to the plasma membrane / Cilium Assembly / Sealing of the nuclear envelope (NE) by ESCRT-III / protein polyglutamylation / left/right pattern formation / regulation of calcineurin-NFAT signaling cascade / sperm axoneme assembly / regulation of microtubule nucleation / Carboxyterminal post-translational modifications of tubulin / 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 / epithelial cilium movement involved in extracellular fluid movement / HSP90 chaperone cycle for steroid hormone receptors (SHR) in the presence of ligand / 9+2 motile cilium / COPI-mediated anterograde transport / cilium movement involved in cell motility / intraciliary transport / regulation of store-operated calcium entry / Kinesins / acrosomal membrane / axoneme assembly / PKR-mediated signaling / cilium movement / left/right axis specification / Aggrephagy / RHO GTPases activate IQGAPs / microtubule sliding / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / Resolution of Sister Chromatid Cohesion / Recycling pathway of L1 / calcium ion sensor activity / The role of GTSE1 in G2/M progression after G2 checkpoint / sperm DNA condensation / COPI-dependent Golgi-to-ER retrograde traffic / axonemal microtubule / Hedgehog 'off' state / RHO GTPases Activate Formins / 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 / Separation of Sister Chromatids / cilium organization / Recruitment of NuMA to mitotic centrosomes / AURKA Activation by TPX2 / gamma-tubulin ring complex / manchette / Regulation of PLK1 Activity at G2/M Transition / 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 / DNA catabolic process / extrinsic component of membrane / nucleoside diphosphate kinase activity / protein targeting to membrane / : / positive regulation of cell motility / GTP biosynthetic process / intermediate filament / Hydrolases; Acting on ester bonds; Exodeoxyribonucleases producing 5'-phosphomonoesters / tubulin complex / protein-serine/threonine phosphatase / AMP binding / ciliary base / regulation of neuron projection development / protein serine/threonine phosphatase activity / receptor clustering / cerebral cortex cell migration / ciliary transition zone / phosphatase activity / mitotic cytokinesis / microtubule organizing center / cellular response to UV-C / cilium assembly / cellular response to unfolded protein / regulation of cell division Similarity search - Function
Spermatogenesis-associated protein 48 / Protein of unknown function DUF4555 / Spermatogenesis-associated protein 48 / Sperm-associated microtubule inner protein 4 / Uncharacterized protein C1orf100 / : / Family of unknown function (DUF5530) / Uncharacterised protein family C20orf85 / Protein of unknown function DUF4562 / Spermatid-specific manchette-related protein 1 ...Spermatogenesis-associated protein 48 / Protein of unknown function DUF4555 / Spermatogenesis-associated protein 48 / Sperm-associated microtubule inner protein 4 / Uncharacterized protein C1orf100 / : / Family of unknown function (DUF5530) / Uncharacterised protein family C20orf85 / Protein of unknown function DUF4562 / Spermatid-specific manchette-related protein 1 / Testis-expressed protein 33 / Testis-expressed sequence 37 protein / CFAP97 domain-containing protein 1/2 / Testis-specific serine/threonine-protein kinase 6, catalytic domain / C10orf82-like / : / : / Normal lung function maintenance, Low in Lung Cancer 1 protein / Domain of unknown function (DUF4562) / Spermatid-specific manchette-related protein 1 / TSC21 family / Testis-expressed sequence 33 protein family / Sperm-associated microtubule inner protein 5 / Ciliary microtubule inner protein 3 / Protein phosphatase 1 regulatory subunit 32 / Testis, prostate and placenta-expressed protein / Testis-expressed protein 49 / Protein phosphatase 1 regulatory subunit 32 / Sperm microtubule inner protein 8 / Sperm microtubule inner protein 11 / : / : / Sperm microtubule inner protein 1 / Protein of unknown function DUF1143 / Atypical dual specificity phosphatase, subfamily B / Protein of unknown function DUF4513 / Cilia- and flagella-associated protein 68 / Sperm-associated microtubule inner protein 10 / Protein of unknown function DUF4541 / Tektin bundle interacting protein 1 / Hemingway/CFA97 / Coiled-coil domain-containing protein 105 / : / Hemingway/CFA97 / Tektin bundle interacting protein 1 / Cilia and flagella associated protein 90 / : / DJBP, EF-hand domain / FAM183 family / EF-hand domain / FAM183A and FAM183B related / Sperm-tail PG-rich repeat / Sperm-associated antigen 8 / Cilia- and flagella-associated protein 95 / NDPK7, first NDPk domain / Cilia- and flagella-associated protein 107 / : / Sperm-tail PG-rich repeat / Protein CFAP95 / Cilia- and flagella-associated protein 143 / Cilia and flagella associated protein 107 / Tektin / Cilia- and flagella-associated protein 276 / : / Tektin family / Protein of unknown function (DUF3695) / Uncharacterised protein FAM166/UPF0605 / Cilia- and flagella-associated protein 77 / Protein Flattop / Ciliary microtubule inner protein 2B-like / Cilia- and flagella-associated protein 77 / Flattop / Nucleoside diphosphate kinase 7 / Piercer of microtubule wall 1/2 / Piercer of microtubule wall 1/2 / Cilia- and flagella-associated protein 141 / Cilia- and flagella-associated protein 141 / Cilia- and flagella-associated protein 161-like domain / : / : / EFHB C-terminal EF-hand domain / : / : / Nucleoside diphosphate kinase 7 N-terminal PH domain / Meiosis-specific nuclear structural protein 1 / Cilia- and flagella-associated protein 45 / NDPK7, second NDPk domain / Cilia- and flagella- associated protein 210 / CFAP53/TCHP / Trichohyalin-plectin-homology domain / Trichohyalin-plectin-homology domain / RIB43A / RIB43A / DM10 domain / EF-hand domain-containing protein EFHC1/EFHC2/EFHB / DM10 domain / DM10 domain profile. / Domains in hypothetical proteins in Drosophila, C. elegans and mammals. Occurs singly in some nucleoside diphosphate kinases. / Enkurin domain / : Similarity search - Domain/homology
Cilia- and flagella-associated protein 77 / Luteinizing hormone beta / Cilia- and flagella- associated protein 210 / Tektin bundle-interacting protein 1 / Protein SPMIP1 / Sperm microtubule inner protein 2 / Tektin-5 / Tubulin alpha-3 chain / Tubulin beta-4B chain / Stabilizer of axonemal microtubules 4 ...Cilia- and flagella-associated protein 77 / Luteinizing hormone beta / Cilia- and flagella- associated protein 210 / Tektin bundle-interacting protein 1 / Protein SPMIP1 / Sperm microtubule inner protein 2 / Tektin-5 / Tubulin alpha-3 chain / Tubulin beta-4B chain / Stabilizer of axonemal microtubules 4 / Tektin-4 / Sperm microtubule inner protein 6 / Sperm-associated antigen 8 / Cilia- and flagella-associated protein 107 / Piercer of microtubule wall 1 protein / Cilia- and flagella-associated protein 52 / Cilia- and flagella-associated protein 144 / Protein SPMIP7 / Meiosis-specific nuclear structural protein 1 / Sperm microtubule inner protein 8 / EF-hand calcium-binding domain-containing protein 6 / Protein Flattop / Cilia- and flagella-associated protein 161 / Enkurin / Tektin-3 / EF-hand calcium-binding domain-containing protein 3 / Cilia- and flagella-associated protein 20 / Sperm microtubule inner protein 11 / EF-hand domain-containing family member B / Ciliary microtubule inner protein 3 / Ciliary microtubule associated protein 1A / Tektin-2 / Testis-specific serine/threonine-protein kinase 6 / Cilia- and flagella-associated protein 95 / Sperm-associated microtubule inner protein 5 / RIB43A-like with coiled-coils protein 1 / Cilia- and flagella-associated protein 68 / Cilia- and flagella-associated protein 53 / EF-hand domain-containing family member C2 / Ciliary microtubule inner protein 2A / Tektin-like protein 1 / RIB43A-like with coiled-coils protein 2 / Sperm-associated microtubule inner protein 4 / Dual specificity phosphatase 21 / Cilia- and flagella-associated protein 141 / Sperm-associated microtubule inner protein 10 / Ciliary microtubule inner protein 4 / EF-hand domain-containing protein 1 / Cilia- and flagella-associated protein 45 / Ciliary microtubule inner protein 1 / Protein SPMIP3 / Cilia- and flagella-associated protein 276 / Protein SPMIP9 / Tektin-1 / Parkin coregulated gene protein homolog / Sperm axonemal maintenance protein CFAP97D1 / Cilia- and flagella-associated protein 90 / Ciliary microtubule inner protein 2C / Nucleoside diphosphate kinase 7 / Piercer of microtubule wall 2 protein Similarity search - Component
Biological species
Mus musculus (house mouse)
Method
single particle reconstruction / cryo EM / Resolution: 3.7 Å
National Natural Science Foundation of China (NSFC)
China
Citation
Journal: Nat Commun / Year: 2026 Title: Doublet microtubule-associated tektins and enzymes differentially regulate sperm flagellar integrity and motility. Authors: Qi Liu / Lunni Zhou / Xiaochen Liang / Pengyu Chen / Bo Li / Shuo Yang / Yuqi Liu / Haibo Zhao / Jin Hu / Shan Feng / Shanshan Xie / Jianping Wu / Miao Gui / Abstract: Doublet microtubule (DMT)-associated proteins assemble and drive sperm flagella, which are essential for successful fertilization. However, the exact roles of different DMT-associated proteins in ...Doublet microtubule (DMT)-associated proteins assemble and drive sperm flagella, which are essential for successful fertilization. However, the exact roles of different DMT-associated proteins in sperm function and the underlying molecular mechanisms remain elusive. Here, we generate four gene-knockout mice based on high-resolution structures targeting distinct DMT components: two intermediate filament-like tektins (TEKT1, TEKT5) and two enzymes (TSSK6, DUSP21). The depletion of TEKT1, shared by sperm flagella and motile cilia, causes male infertility characterized by impaired sperm motility and loss of the tektin bundle, whereas sperm-specific Tekt5 knockout (KO) mice remain fertile with largely normal flagellar function, indicating functional divergence within the tektin family. Tssk6 KO spermatozoa exhibit severely disturbed morphology and motility, resulting in homozygote infertility and heterozygote subfertility. Phosphoproteomics reveals dysregulated phosphorylation of axonemal proteins, highlighting the critical role of kinase-mediated signaling in regulating sperm motility. Conversely, Dusp21 KO mice display no fertility or sperm motility defects, suggesting compensatory phosphatase activity. Phenotypic comparisons between Tekt1 and Tssk6 KO mice suggest their involvement in distinct subtypes of asthenozoospermia. Overall, this study elucidates how filamentous and enzymatic DMT proteins govern sperm function through divergent mechanisms, which have implications for molecular diagnosis of male infertility.
Name: Tubulin alpha-3 chain / type: protein_or_peptide / ID: 8 / Number of copies: 167 / Enantiomer: LEVO EC number: Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement
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