| 2007 |
SEPT12 was identified as a novel septin via yeast two-hybrid screen using SEPT5 as bait; it purifies with bound nucleotide, binds phosphoinositides, and co-localizes with SEPT4 into robust curved filaments when co-expressed in CHO cells, placing it at the sperm annulus. |
Yeast two-hybrid screen, nucleotide binding assay, phosphoinositide binding assay, co-expression/co-localization in CHO cells, RT-PCR, Northern blot, Western blot |
Cell motility and the cytoskeleton |
Medium |
17685441
|
| 2007 |
SEPT12 interacts with SEPT6 in vitro and in vivo (co-IP); co-expression of SEPT12 alters the filamentous structure of SEPT6 in HeLa cells. SEPT12 localizes to central spindle and midbody during anaphase and cytokinesis. |
Co-immunoprecipitation (in vitro and in vivo), co-expression in HeLa cells, immunocytochemistry |
Journal of biochemistry and molecular biology |
Medium |
18047794
|
| 2008 |
SEPT12 binds GTP in vitro; a G56N mutation in the GTP-binding motif abolishes binding. GTP binding is required for filament formation (G56N forms aggregates instead of filaments) and for interaction with SEPT11, but not for self-interaction. |
In vitro GTP-binding assay, site-directed mutagenesis, immunocytochemistry in HeLa cells |
Molecules and cells |
High |
18443421
|
| 2012 |
Two missense mutations in the GTPase domain (T89M and D197N) identified in infertile men: T89M significantly reduces GTP hydrolytic activity; D197N interferes with GTP binding. Both mutant proteins restrict wild-type SEPT12 filament formation in a dose-dependent dominant-negative manner, disrupting sperm annulus integrity. |
GTP hydrolysis assay, GTP binding assay, filament formation assay, in silico structural modeling |
Human mutation |
High |
22275165
|
| 2012 |
A c.474G>A splice-site variant causes exon 5 skipping, producing a truncated SEPT12 lacking the C-terminal half; this truncated protein inhibits wild-type SEPT12 filament formation in a dose-dependent manner (ex vivo), and is associated with bent tail and nuclear DNA damage in sperm. |
DNA sequencing, RT-PCR splice assay, filament formation assay (ex vivo), immunocytochemistry |
PloS one |
Medium |
22479503
|
| 2013 |
SEPT12 interacts with α- and β-tubulins by co-IP; SEPT12 co-localizes with tubulins during spermiogenesis in transgenic mice. shRNA-mediated loss of SEPT12 disrupts α- and β-tubulin organization and impairs sperm head morphogenesis and tail elongation. |
Co-immunoprecipitation, SEPTIN12-transgenic mouse model, shRNA knockdown, confocal microscopy |
International journal of molecular sciences |
Medium |
24213608
|
| 2015 |
SEPT12 assembles octameric filaments with SEPT7, SEPT6, SEPT2, and SEPT4 (arrangements 12-7-6-2-2-6-7-12 or 12-7-6-4-4-6-7-12) at the sperm annulus. The GTP-binding domain of SEPT12 is required for interaction with SEPT7, and N- and C-termini are required for SEPT12 self-polymerization into filaments. The D197N knock-in mouse shows disorganized annulus, bent tail, reduced sperm motility, and loss of SEPT ring structure. |
Co-immunoprecipitation, deletion/domain mapping, knock-in mouse model (D197N), electron microscopy, immunofluorescence |
Journal of cell science |
High |
25588830
|
| 2015 |
SEPT12 interacts with SPAG4 (SUN4) in a male germ cell line (co-IP). SEPT12, SPAG4, and LAMINB1 form complexes at the nuclear periphery of round spermatids. A SEPT12 disease mutation disrupts these nuclear envelope complexes (co-IP). SEPT12 links SEPT proteins to the SUN/LAMIN nuclear envelope complex during postmeiotic germ cell development. |
Yeast two-hybrid, co-immunoprecipitation, co-localization by confocal microscopy in germ cell line and human spermatogenic cells |
PloS one |
Medium |
25775403
|
| 2016 |
SEPT12 interacts with NDC1 (nuclear pore complex protein) in male germ cell line (co-IP). NDC1 overexpression restricts SEPT12 localization to the nucleus and represses SEPT12 filament formation. In D197N mutant sperm, NDC1 disperses from the sperm neck to manchette region and annulus. |
Yeast two-hybrid, co-immunoprecipitation, overexpression/localization in germ cell line, knock-in mouse model |
International journal of molecular sciences |
Medium |
27854341
|
| 2018 |
SEPT12 phosphorylation on Ser198 by Protein Kinase A (PKA) regulates sperm annulus architecture. A phosphomimetic SEPT12 S198 knock-in mouse shows poor male fertility, weak sperm motility, and loss of the sperm annulus. Phosphorylation at Ser198 interferes with SEPT12 polymerization into complexes and filaments. |
Knock-in mouse model (phosphomimetic), fertility assays, sperm motility analysis, filament polymerization assay, PKA kinase identification |
Cytoskeleton (Hoboken, N.J.) |
High |
30160375
|
| 2018 |
CDC42 negatively regulates SEPT12 polymerization. Wild-type CDC42 and constitutively active CDC42Q61L substantially repress SEPT12 polymerization, whereas dominant-negative CDC42T17N does not. CDC42 and SEPT12 co-localize in perinuclear manchette and midpiece regions of spermatids/spermatozoa. |
Ectopic expression analysis with CDC42 mutants, filament formation assay, scanning electron microscopy, co-localization by immunofluorescence |
International journal of molecular sciences |
Medium |
30189608
|
| 2018 |
The SEPT12 gene promoter is a 245 bp region upstream of the transcription start site. Androgen receptor (AR) and estrogen receptor α (ERα) directly bind this region (confirmed by chromatin immunoprecipitation). Truncation of AR or ERα binding sites decreases promoter activity; treatment with 17β-estradiol or 5α-dihydrotestosterone enhances promoter activity. |
Promoter deletion analysis, chromatin immunoprecipitation (ChIP), luciferase reporter assay |
Biochimie |
Medium |
30513371
|
| 2019 |
SEPT12 expression alters nuclear membrane localization of SPAG4 (SUN4) and also alters localization of LAMINA/C; this effect is specific to SEPT12 and not observed with SEPT1, SEPT6, SEPT7, or SEPT11. |
Confocal microscopy after SEPT12 overexpression in testicular cancer cell line, comparison with other septins |
International journal of molecular sciences |
Medium |
30866452
|
| 2020 |
SEPT12 forms a complex with SEPT1, SEPT2, SEPT10, and SEPT11 at the sperm neck. The D197N mutation disrupts this complex and causes defective connecting pieces (head-tail junction). SEPT12 interacts and co-localizes with γ-tubulin in elongating spermatids, suggesting SEPT12 and pericentriolar materials jointly form connecting pieces. |
Co-immunoprecipitation, electron microscopy, immunofluorescence, D197N knock-in mouse model |
Molecular human reproduction |
High |
32392324
|
| 2022 |
Homozygous Septin12 knockout (but not heterozygous) male mice are infertile with reduced sperm counts and abnormal morphology. Loss of PLCζ around the acrosome in Septin12-null sperm is identified as the likely mechanism for fertilization failure, which can be rescued by artificial oocyte activation (AOA). |
Septin12 knockout mouse model, ICSI with AOA, immunofluorescence for PLCζ localization |
Frontiers in cell and developmental biology |
Medium |
35547809
|
| 2024 |
SUN5 interacts with SEPT12 (and LAMINB1) in mouse testis, identified by immunoprecipitation-mass spectrometry and validated by further studies. The SUN5-SEPT12 interaction promotes their co-aggregation at the sperm neck; Sun5 deficiency disrupts the LaminB1/SUN5/SEPT12 complex, causing separation of the SEPT12-proximal centriole from the nucleus and breakage of the head-to-tail junction. |
Immunoprecipitation-mass spectrometry, co-immunoprecipitation, immunofluorescence, Sun5 knockout mouse model |
Molecular human reproduction |
High |
38870534
|
| 2025 |
A novel SEPT12 T96I mutation causes male infertility; overexpression of SEPT12 T96I in NT2/D1 cells impairs SEPT7 filament formation, demonstrating that SEPT12 filament integrity is required for proper SEPT7 filament organization. Sperm from T96I carriers show obliquely positioned annulus with SEPT12/SEPT7 co-localization defects, large nuclear vacuoles, and acrosomal decondensation. |
Immunofluorescence, transmission electron microscopy, overexpression in NT2/D1 cells |
Frontiers in cell and developmental biology |
Medium |
39850804
|