Affinage

ODF1

Outer dense fiber protein 1 · UniProt Q14990

Length
250 aa
Mass
28.4 kDa
Annotated
2026-06-10
31 papers in source corpus 17 papers cited in narrative 17 extracted findings
Cross-family judge vs UniProt: Affinage preferred faithfulness: 6/6 claims corpus-supported (100%)

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

ODF1 (HSPB10) is a spermatid-specific structural protein that forms a core component of the sperm tail outer dense fibers and mediates the mechanical coupling of the sperm head to the flagellum (PMID:1699827, PMID:22037768). It self-associates through an N-terminal region containing a leucine-zipper-like motif and uses leucine zippers to nucleate a network of sperm tail interactions, binding the major outer dense fiber protein ODF2 as well as SPAG4, SPAG5, OIP1, CCDC42, and WDR64 (PMID:7521678, PMID:9045620, PMID:10373309, PMID:11468777, PMID:12533418, PMID:31475146, PMID:39386799). Several of these partners are recruited to or localize at the manchette and connecting piece, positioning ODF1 within the apparatus that builds and links the sperm neck and tail (PMID:10373309, PMID:31475146, PMID:39386799). ODF1 is post-translationally regulated: Cdk5/p35 phosphorylates ODF1 at Ser193 in its C-terminal Cys-X-Pro region, enhancing binding to the RING-finger E3 ligase candidate OIP1 (PMID:17762160). Knockout and haplo-deficiency studies establish that ODF1 is dispensable for assembling the connecting piece but is required to stabilize the head-to-tail linkage; its loss causes sperm decapitation, mitochondrial sheath and outer dense fiber disorganization, and male infertility in mice (PMID:22037768, PMID:25118300, PMID:36142191). Odf1 expression is controlled at multiple levels — transcriptionally by the testis-specific factor TTF-D and a CREB-like factor at its promoter (PMID:1731343, PMID:10206985), and translationally through storage of its mRNA in inactive ribonucleoprotein particles with later activation, including TENT5C-mediated poly(A) tail extension required for ODF1 accumulation at the spermatid neck (PMID:8873064, PMID:42009655).

Mechanistic history

Synthesis pass · year-by-year structured walk · 17 steps
  1. 1990 Medium

    Established the molecular identity of ODF1 as a small spermatid-specific protein bearing a leucine-zipper-like N-terminal helix, framing it as a candidate structural/interaction protein of the developing sperm tail.

    Evidence Differential cDNA cloning and sequencing in rat early spermatids

    PMID:1699827

    Open questions at the time
    • Subcellular localization and binding partners not yet defined
    • Function of the leucine-zipper motif untested
  2. 1992 High

    Defined how the cell-type-restricted expression of Odf1 is achieved, identifying promoter elements bound by a testis-specific factor and a CREB-like factor.

    Evidence In vitro transcription, DNase I footprinting and EMSA with seminiferous epithelium extracts

    PMID:1731343

    Open questions at the time
    • Molecular identity of TTF-D not yet resolved
    • In vivo requirement of these elements untested
  3. 1994 Medium

    Showed that ODF1 localizes to the sperm tail and self-associates via its N-terminal half, supporting a structural role in outer dense fibers.

    Evidence Immunofluorescence and reciprocal co-IP with deletion mapping

    PMID:7521678

    Open questions at the time
    • Stoichiometry and higher-order assembly unknown
    • Single-lab data
  4. 1996 Medium

    Revealed that ODF1 output is gated translationally, with mRNA stored in inactive RNP particles and translated during the maturation phase as ODF diameter increases.

    Evidence In situ hybridization, immunohistochemistry and polysome fractionation

    PMID:8873064

    Open questions at the time
    • Molecular machinery controlling the storage-to-translation switch not identified
  5. 1997 High

    Identified ODF2 as a direct leucine-zipper partner of ODF1, defining the core protein-protein contact of the outer dense fiber.

    Evidence Yeast two-hybrid with leucine-to-alanine mutagenesis and immunoelectron microscopy

    PMID:9045620

    Open questions at the time
    • In vivo assembly hierarchy of ODF1/ODF2 not established
  6. 1999 High

    Expanded the ODF1 interactome with SPAG4 (manchette/axoneme) and connected the protein to candidate tail-protein transport.

    Evidence Yeast two-hybrid, co-IP and immunofluorescence

    PMID:10373309

    Open questions at the time
    • Functional consequence of SPAG4 binding untested in vivo
  7. 1999 Medium

    Characterized the DNA-binding behavior of the testis-specific factor TTF-D, showing it binds both ds and ss Odf1 promoter sites and activates transcription.

    Evidence EMSA and promoter-reporter assays with ss/ds oligonucleotide competition

    PMID:10206985

    Open questions at the time
    • Identity of TTF-D peptides not cloned
    • Mechanism of ss vs ds binding unresolved
  8. 2001 Medium

    Added SPAG5 as a leucine-zipper ODF1 partner expressed across spermatocytes and spermatids.

    Evidence Yeast two-hybrid with deletion mutagenesis

    PMID:11468777

    Open questions at the time
    • No co-IP or localization confirmation in this study
    • Functional role undetermined
  9. 2003 Medium

    Linked ODF1 to a potential ubiquitin-mediated regulatory pathway by identifying the RING-finger protein OIP1 binding ODF1's conserved C-terminal Cys-Gly-Pro repeats.

    Evidence Yeast two-hybrid, deletion mutagenesis and immunofluorescence

    PMID:12533418

    Open questions at the time
    • E3 ligase activity of OIP1 toward ODF1 not demonstrated
    • No degradation assay
  10. 2007 High

    Connected a kinase signal to the ODF1-OIP1 axis, showing Cdk5/p35 phosphorylates ODF1 at Ser193 to enhance OIP1 binding, implying regulated turnover.

    Evidence In vitro kinase assay, mass spectrometry phosphosite mapping, phospho-mutant binding assays

    PMID:17762160

    Open questions at the time
    • Functional consequence of phospho-enhanced OIP1 binding in vivo not shown
    • ODF1 ubiquitination/degradation not directly demonstrated
  11. 2011 High

    Established ODF1's essential physiological role: it is required for tight head-to-tail linkage and proper mitochondrial sheath/ODF organization, with knockout males infertile from decapitation.

    Evidence Knockout mouse, transmission electron microscopy, motility assays

    PMID:22037768

    Open questions at the time
    • Molecular basis of mechanical coupling not resolved
    • Whether defect is assembly or maintenance unclear at this stage
  12. 2014 Medium

    Demonstrated dose sensitivity of the linkage, with haplo-deficiency widening the nuclear membrane-to-capitulum distance and reducing fecundity despite normal routine semen parameters.

    Evidence Targeted disruption with backcrossing and ultrastructural analysis

    PMID:25118300

    Open questions at the time
    • Single genetic background
    • Mechanism linking ODF1 dosage to coupling strength unknown
  13. 2019 Medium

    Placed ODF1 in a connecting-piece/manchette network by showing CCDC42 binds both ODF1 and ODF2 yet localizes independently of ODF1.

    Evidence Co-IP and immunofluorescence in germ and somatic cells

    PMID:31475146

    Open questions at the time
    • Functional interdependence of CCDC42 and ODF1 not tested
  14. 2019 Medium

    Challenged the strict testis-exclusivity of ODF1 by detecting it in rat kidney collecting ducts.

    Evidence Western blot, immunofluorescence, RT-PCR, mass spectrometry

    PMID:31867458

    Open questions at the time
    • Function of ODF1 in kidney unknown
    • Not confirmed in other species
  15. 2022 Medium

    Resolved the assembly-versus-maintenance question, showing the connecting piece forms normally without ODF1 but fails mechanically under force, defining ODF1 as a stabilizer rather than a builder.

    Evidence TEM of developing spermatids from knockout mice

    PMID:36142191

    Open questions at the time
    • Molecular nature of the stabilizing interaction unidentified
  16. 2024 Medium

    Added WDR64 as an ODF1 complex partner spanning the manchette and midpiece, implicating the complex in nucleus shaping and flagellum assembly.

    Evidence IP-MS and immunofluorescence co-localization

    PMID:39386799

    Open questions at the time
    • Single method for interaction
    • In vivo requirement of WDR64-ODF1 complex untested
  17. 2026 High

    Defined an upstream translational control mechanism: TENT5C-mediated poly(A) extension of Odf1 mRNA is required for ODF1 accumulation at the spermatid neck, with loss phenocopying ODF1 deficiency.

    Evidence Catalytically inactive TENT5C knock-in mouse, transcriptome poly(A) profiling, Western blot and immunofluorescence

    PMID:42009655

    Open questions at the time
    • How TENT5C is targeted specifically to Odf1 mRNA unknown
    • Direct vs indirect effect on ODF1 not fully separated

Open questions

Synthesis pass · forward-looking unresolved questions
  • It remains unknown whether the Cdk5/OIP1 phosphorylation-degradation model operates in vivo, and what molecular interaction physically transmits the stabilizing force at the head-tail junction.
  • No in vivo demonstration of OIP1-mediated ODF1 degradation
  • Biophysical mechanism of mechanical stabilization unresolved
  • Function of non-testicular ODF1 unaddressed

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005198 structural molecule activity 3
Localization
GO:0005856 cytoskeleton 2 GO:0005815 microtubule organizing center 1
Pathway
R-HSA-1474165 Reproduction 2
Partners
CCDC42CDK5ODF2OIP1SPAG4SPAG5WDR64
Complex memberships
WDR64/ODF1 complexsperm tail outer dense fibers

Evidence

Reading pass · 17 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
1990 RT7 (ODF1) encodes a putative 90-amino acid protein with an N-terminal amphipathic alpha helix containing a leucine zipper-like structure; it is specifically expressed at very high levels in rat early spermatids and not detectable in any other tissue. Differential cDNA cloning, nucleotide sequencing, transcription start site mapping Developmental biology Medium 1699827
1992 The Odf1 promoter is positively regulated by a testis-specific transcription factor TTF-D, a factor similar to CREB, and a nuclear factor binding immediately upstream of the transcription start site, as demonstrated in an in vitro transcription system derived from rat seminiferous epithelium. In vitro transcription assay (seminiferous epithelium nuclear extracts), DNase I footprinting, gel retardation (EMSA) Proceedings of the National Academy of Sciences of the United States of America High 1731343
1994 ODF1 (RT7) protein localizes to the sperm tail and self-associates through a region in the N-terminal half, forming stable complexes with itself, consistent with it being a structural component of sperm tail outer dense fibers. Immunofluorescence microscopy, confocal laser immunofluorescence, co-immunoprecipitation with deletion analysis Biology of reproduction Medium 7521678
1996 Odf1 mRNA is transcribed from step 6 round spermatids through step 17, but the bulk is stored in translationally inactive ribonucleoprotein particles; translation is greatly enhanced in the maturation phase of spermiogenesis coinciding with marked increase in outer dense fiber diameter. In situ hybridization, immunohistochemistry, polysome fractionation on sucrose gradients, Northern blot Molecular reproduction and development Medium 8873064
1997 ODF1 (Odf27) interacts with the major 84-kDa outer dense fiber protein ODF2 (Odf84) via leucine zippers; all leucine residues in the upstream leucine zipper of ODF2 are required for interaction with ODF1. Yeast two-hybrid screen with ODF1 leucine zipper as bait, fluorescence microscopy, immunoelectron microscopy, leucine-to-alanine mutagenesis The Journal of biological chemistry High 9045620
1999 SPAG4 (a 49-kDa spermatid-specific protein) interacts specifically with ODF1 but not ODF2 via a leucine zipper, and localizes to the manchette and axoneme of spermatids, suggesting a role in protein transport to sperm tail structures. Yeast two-hybrid cloning using ODF1 as bait, co-immunoprecipitation, immunofluorescence microscopy Developmental biology High 10373309
1999 TTF-D, a testis-specific nuclear factor containing three peptides (22, 25, and 35 kDa), binds both double-stranded and single-stranded DNA at cognate sites in the Odf1 and c-mos promoters and activates Odf1 promoter activity; both ds and ss binding site oligonucleotides can specifically repress Odf1 promoter activity. Electrophoretic mobility shift assay (EMSA), promoter-reporter transcription assays, competition with single-stranded and double-stranded oligonucleotides The Journal of biological chemistry Medium 10206985
2001 SPAG5, a 200-kDa testicular protein with similarity to the mitotic spindle protein Deepest, interacts strongly with ODF1 via a leucine zipper in its C-terminal region; SPAG5 is expressed in pachytene spermatocytes and spermatids. Yeast two-hybrid cloning using ODF1 as bait, deletion mutagenesis of leucine zipper Molecular reproduction and development Medium 11468777
2003 OIP1, a novel RING finger protein (H2 subclass), binds specifically to evolutionarily conserved Cys-Gly-Pro repeats in the C-terminus of ODF1; deletion of the RING motif significantly decreases ODF1 binding, and OIP1 localizes to the sperm tail. Yeast two-hybrid screen using ODF1 N-terminal leucine zipper as bait, deletion mutagenesis, immunofluorescence localization Biology of reproduction Medium 12533418
2007 Cdk5/p35 phosphorylates ODF1 specifically at Ser193 in the C-terminal Cys-X-Pro region, and this phosphorylation enhances interaction between ODF1 and the RING finger E3 ubiquitin ligase candidate OIP1; ODF2 interacts with Cdk5 and p35 individually but not with the Cdk5/p35 heterodimer, and Cdk5/p35 does not phosphorylate ODF2. In vitro kinase assay, mass spectrometry (phosphosite identification), co-immunoprecipitation with deletion mutants, binding assays with phospho-mimetic/phospho-null mutants Cellular physiology and biochemistry High 17762160
2011 ODF1/HSPB10 is essential for tight linkage of the sperm head to the tail and for correct organization of the mitochondrial sheath and outer dense fibers; Odf1-null male mice are infertile due to sperm decapitation, while heterozygous males are fertile but show reduced sperm motility. Targeted gene disruption (knockout mouse), transmission electron microscopy, sperm motility assays Molecular and cellular biology High 22037768
2014 Haplo-deficiency of ODF1 in incipient congenic (129/Sv background) mice causes an enlargement of the distance between the nuclear membrane and capitulum, indicating weakening of sperm head-to-tail coupling, with severe reduction in male fecundity without detectable abnormal sperm parameters by routine semen analysis. Targeted gene disruption, successive backcrossing, ultrastructural analysis by transmission electron microscopy Reproduction (Cambridge, England) Medium 25118300
2019 CCDC42 interacts with both ODF1 and ODF2 in male germ cells and localizes to the manchette, connecting piece (HTCA), and sperm tail; in somatic cells CCDC42 localizes to the centrosome, and its centrosomal/sperm tail localization appears independent of ODF1. Co-immunoprecipitation, immunofluorescence microscopy in spermatids and somatic cells Frontiers in cell and developmental biology Medium 31475146
2019 ODF1 protein (and its mRNA) is expressed in rat kidney collecting ducts, demonstrating that ODF1 is not exclusively a testicular/sperm protein. Western blot, immunofluorescence, RT-PCR, proteomic analysis by mass spectrometry Heliyon Medium 31867458
2022 ODF1 is dispensable for the structural development of the connecting piece (which develops normally in ODF1-deficient spermatids by TEM), but is required for stable mechanical linkage of sperm head and tail; decapitation occurs when force is applied, indicating a maintenance/stabilization rather than assembly role. Transmission electron microscopy of developing spermatids from Odf1 knockout mice International journal of molecular sciences Medium 36142191
2024 WDR64 (a testis-specific WD40 repeat protein) interacts with ODF1 to form a protein complex; the WDR64/ODF1 complex localizes to the manchette during nucleus shaping and to the midpiece of mature spermatozoa, implicating it in manchette and flagellum assembly. Immunoprecipitation–mass spectrometry (IP-MS), immunofluorescence microscopy Heliyon Medium 39386799
2026 TENT5C poly(A) polymerase extends the poly(A) tail of Odf1 mRNA in spermatids; in the absence of TENT5C catalytic activity, Odf1 transcripts have shorter poly(A) tails, ODF1 protein fails to accumulate at the spermatid neck, and mice produce headless spermatozoa with flagellar abnormalities phenocopying ODF1 deficiency. Poly(A) tail profiling across transcriptome, Western blot, immunofluorescence localization of ODF1 protein, catalytically inactive TENT5C knock-in mouse model Nature communications High 42009655

Source papers

Stage 0 corpus · 31 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
1999 Spag4, a novel sperm protein, binds outer dense-fiber protein Odf1 and localizes to microtubules of manchette and axoneme. Developmental biology 134 10373309
2011 The small heat shock protein ODF1/HSPB10 is essential for tight linkage of sperm head to tail and male fertility in mice. Molecular and cellular biology 120 22037768
1997 Interactional cloning of the 84-kDa major outer dense fiber protein Odf84. Leucine zippers mediate associations of Odf84 and Odf27. The Journal of biological chemistry 86 9045620
1990 A new rat gene RT7 is specifically expressed during spermatogenesis. Developmental biology 65 1699827
1990 RT7-defined alloantigens in rats are part of the leucocyte common antigen family. Scandinavian journal of immunology 55 2192438
1992 Factors involved in regulation of the RT7 promoter in a male germ cell-derived in vitro transcription system. Proceedings of the National Academy of Sciences of the United States of America 52 1731343
2001 Testicular protein Spag5 has similarity to mitotic spindle protein Deepest and binds outer dense fiber protein Odf1. Molecular reproduction and development 51 11468777
2019 CCDC42 Localizes to Manchette, HTCA and Tail and Interacts With ODF1 and ODF2 in the Formation of the Male Germ Cell Cytoskeleton. Frontiers in cell and developmental biology 50 31475146
2014 Haplo-deficiency of ODF1/HSPB10 in mouse sperm causes relaxation of head-to-tail linkage. Reproduction (Cambridge, England) 44 25118300
1984 Antiidiotypic immunity in interstitial nephritis. II. Rats developing anti-tubular basement membrane disease fail to make an antiidiotypic regulatory response: the modulatory role of an RT7.1+, OX8- suppressor T cell mechanism. The Journal of experimental medicine 41 6231352
1996 Transcription and translation of the outer dense fiber gene (Odf1) during spermiogenesis in the rat. A study by in situ analyses and polysome fractionation. Molecular reproduction and development 27 8873064
2019 Roseltide rT7 is a disulfide-rich, anionic, and cell-penetrating peptide that inhibits proteasomal degradation. The Journal of biological chemistry 24 31727740
2002 In vivo depletion of hematopoietic stem cells in the rat by an anti-CD45 (RT7) antibody. Blood 23 11986209
1998 A novel testis-specific gene, SPAG4, whose product interacts specifically with outer dense fiber protein ODF27, maps to human chromosome 20q11.2. Cytogenetics and cell genetics 22 9691178
1994 Testis-specific RT7 protein localizes to the sperm tail and associates with itself. Biology of reproduction 22 7521678
1987 Monoclonal antibodies to RT7 and LCA antigens in the rat: cell distribution and segregation analysis. Hybridoma 21 2960606
1995 Sequence of mouse Odf1 cDNA and its chromosomal localization: extension of the linkage group between human chromosome 8 and mouse chromosome 15. Cytogenetics and cell genetics 20 7789171
2007 ODF1 phosphorylation by Cdk5/p35 enhances ODF1-OIP1 interaction. Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology 19 17762160
2003 Novel RING finger protein OIP1 binds to conserved amino acid repeats in sperm tail protein ODF1. Biology of reproduction 16 12533418
2019 A CRTH2 antagonist, CT-133, suppresses NF-κB signalling to relieve lipopolysaccharide-induced acute lung injury. European journal of pharmacology 12 30951719
2009 [Differential expression of ODF1 in human ejaculated spermatozoa and its clinical significance]. Zhonghua nan ke xue = National journal of andrology 10 20112736
1999 Testis-specific TTF-D binds to single-stranded DNA in the c-mos and Odf1 promoters and activates Odf1. The Journal of biological chemistry 10 10206985
2022 Development of the Connecting Piece in ODF1-Deficient Mouse Spermatids. International journal of molecular sciences 9 36142191
2008 Comparative genomics reveals gene-specific and shared regulatory sequences in the spermatid-expressed mammalian Odf1, Prm1, Prm2, Tnp1, and Tnp2 genes. Genomics 9 18562159
2001 Long-term allograft acceptance induced by single dose anti-leukocyte common antigen (RT7) antibody in the rat. Transplantation 9 11374414
2020 Overexpression of ODF1 in Gastrointestinal Tract Neuroendocrine Neoplasms: a Novel Potential Immunohistochemical Biomarker for Well-differentiated Neuroendocrine Tumors. Endocrine pathology 8 32869188
2019 ODF1, sperm flagelar protein is expressed in kidney collecting ducts of rats. Heliyon 7 31867458
2018 CRTH2 antagonist, CT‑133, effectively alleviates cigarette smoke-induced acute lung injury. Life sciences 6 30468833
2020 Lipid-Embedded Molecular Dynamics Simulation Model for Exploring the Reverse Prostaglandin D2 Agonism of CT-133 towards CRTH2 in the Treatment of Type-2 Inflammation Dependent Diseases. Chemistry & biodiversity 5 32034875
2024 WDR64, a testis-specific protein, is involved in the manchette and flagellum formation by interacting with ODF1. Heliyon 3 39386799
2026 TENT5C extends Odf1 poly(A) tail to sustain sperm morphogenesis and fertility. Nature communications 1 42009655

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