# Proteome of oocyte spindle identifies Ccdc69 regulates spindle assembly like “band-tightening spell”

**Authors:** Jia-Ni Guo, Liu Zhu, Tie-Gang Meng, Yi-Na Zhang, Si-Min Sun, Xue-Mei Yang, Bing-Wang Zhao, Yi-Ke Lu, Yuan-Hong Xu, Wei Yue, Zhiming Han, Catherine C. L. Wong, Zhen-Bo Wang

PMC · DOI: 10.1007/s00018-025-05821-7 · Cellular and Molecular Life Sciences: CMLS · 2025-07-30

## TL;DR

This study identifies Ccdc69 as a new protein that helps form the spindle during mouse oocyte maturation, similar to a magical tightening band from a Chinese classic.

## Contribution

The study introduces Ccdc69 as a novel regulator of spindle assembly in mouse oocytes.

## Key findings

- Ccdc69 knockout oocytes had elongated MI spindles.
- Overexpression of Ccdc69 disrupted microtubule formation and aMTOCs distribution.
- Ccdc69 overexpression impaired K-MT attachment and delayed meiotic progression.

## Abstract

Meiotic spindle is an intricate structure and required for chromosome segregation and the proper meiotic progression during oocyte maturation, and its function is regulated by a complex network of proteins located at spindle and its peripheral region. However, proteome of meiotic spindle remains poorly characterized. Here, we acquired the proteomic profile of spindles isolated from metaphase I (MI) and metaphase II (MII) mouse oocytes. In particular, we identified Ccdc69 as a novel regulator of spindle assembly in mouse oocytes. Although deletion of Ccdc69 did not affect female fertility, the MI spindles were elongated in Ccdc69 knockout oocytes. Overexpression of Ccdc69 induced spindle defects by reducing microtubule formation and disturbing acentriolar microtubule organization centers (aMTOCs) distribution. Furthermore, Ccdc69 overexpression impaired kinetochore-microtubule (K-MT) attachment and delayed meiotic progression by abnormal activation of spindle assembly checkpoint (SAC). Taken together, our study depicts the proteome of spindles during mouse oocyte maturation and demonstrates that Ccdc69 regulates spindle assembly and meiotic progression the way similar to “The Tightening Spell of Sun Wukong’s Golden Headband” in the famous Chinese Classic Journey to the West.

The online version contains supplementary material available at 10.1007/s00018-025-05821-7.

## Linked entities

- **Genes:** CCDC69 (coiled-coil domain containing 69) [NCBI Gene 26112]
- **Species:** Mus musculus (taxon 10090)

## Full-text entities

- **Genes:** Aurka (aurora kinase A) [NCBI Gene 20878] {aka AIRK1, ARK-1, Ark1, Aurora-A, Ayk1, IAK}, Aurkb (aurora kinase B) [NCBI Gene 20877] {aka AIM-1, AIRK2, Aik2, Aim1, Ark2, AurB}, Pttg1 (pituitary tumor-transforming gene 1) [NCBI Gene 30939] {aka Pttg, Pttg3}, Ran (RAN, member RAS oncogene family) [NCBI Gene 19384], Myc (Myc proto-oncogene, bHLH transcription factor) [NCBI Gene 17869] {aka Myc2, Niard, Nird, bHLHe39}, Plk4 (polo like kinase 4) [NCBI Gene 20873] {aka 1700028H20, Sak, Stk18}, Kif2a (kinesin family member 2A) [NCBI Gene 16563] {aka C530030B14Rik, Kif2, Kns2, M-kinesin}, Kif2c (kinesin family member 2C) [NCBI Gene 73804] {aka 4930402F02Rik, ESTM5, Knsl6, MCAK}, Cdc20 (cell division cycle 20) [NCBI Gene 107995] {aka 2310042N09Rik, p55CDC}, BUB3 (BUB3 mitotic checkpoint protein) [NCBI Gene 9184] {aka BUB3L, hBUB3}, Pcnt (pericentrin (kendrin)) [NCBI Gene 18541] {aka KEN, Pcnt2, kendrin, m239Asp, m275Asp}, Ndc80 (NDC80 kinetochore complex component) [NCBI Gene 67052] {aka 2610020P18Rik, HEC, HEC1, Kntc2}, Bub1b (BUB1B, mitotic checkpoint serine/threonine kinase) [NCBI Gene 12236] {aka BUBR1}, Ccdc69 (coiled-coil domain containing 69) [NCBI Gene 52570] {aka 2210021E03Rik, D11Ertd461e}, Cga (glycoprotein hormones, alpha subunit) [NCBI Gene 12640] {aka CG-alpha, FSHA, GPHA1, GPHalpha, HCG, LHA}, TACC3 (transforming acidic coiled-coil containing protein 3) [NCBI Gene 10460] {aka ERIC-1, ERIC1, Tacc4, maskin}, Incenp (inner centromere protein) [NCBI Gene 16319] {aka 2700067E22Rik, C130081E20}, Kif11 (kinesin family member 11) [NCBI Gene 16551] {aka Eg5, Kif8, Kifl1, Knsl1}, Cenpe (centromere protein E) [NCBI Gene 229841] {aka 312kDa, C530022J18, CENP-E, Kif10}, TPX2 (TPX2 microtubule nucleation factor) [NCBI Gene 22974] {aka C20orf1, C20orf2, DIL-2, DIL2, FLS353, GD:C20orf1}, Mtus1 (mitochondrial tumor suppressor 1) [NCBI Gene 102103] {aka ATBP135, Atip1, B430010I23Rik, B430305I03Rik, Cctsg1-440, MD44}, Tpx2 (TPX2, microtubule-associated) [NCBI Gene 72119] {aka 2610005B21Rik, DIL2, REPP86, p100}, Pbrm1 (polybromo 1) [NCBI Gene 66923] {aka 2610016F04Rik, BAF180, Pb1}, Actb (actin, beta) [NCBI Gene 11461] {aka Actx, E430023M04Rik, beta-actin}, Bub3 (BUB3 mitotic checkpoint protein) [NCBI Gene 12237], Tacc3 (transforming, acidic coiled-coil containing protein 3) [NCBI Gene 21335] {aka Aint, Eric1}, Birc5 (baculoviral IAP repeat-containing 5) [NCBI Gene 11799] {aka AAC-11, Api4, TIAP, survivin40}, AURKA (aurora kinase A) [NCBI Gene 6790] {aka AIK, ARK1, AURA, BTAK, PPP1R47, STK15}, CCDC69 (coiled-coil domain containing 69) [NCBI Gene 26112], Rb1cc1 (RB1-inducible coiled-coil 1) [NCBI Gene 12421] {aka 2900055E04Rik, 5930404L04Rik, Cc1, FIP200, LaXp180}, Mtus2 (microtubule associated tumor suppressor candidate 2) [NCBI Gene 77521] {aka 5730592G18Rik, A730013O20Rik, C130038G02Rik, Cazip, Gm763, mKIAA0774}, Cat4 (dominant cataract 4) [NCBI Gene 109738] {aka Anc, Apc-1, Apcat-1, Apcat1, Apyc, Pcs-2}
- **Diseases:** aneuploidy (MESH:D000782), tumors (MESH:D009369), SAC (MESH:D002277), MII (MESH:C537730), WT (MESH:D009396), MI (MESH:D006969), GVBD (MESH:D054331)
- **Chemicals:** PBS (MESH:D007854), H2O (MESH:D014867), Triton X-100 (MESH:D017830), CO2 (MESH:D002245), Cy5 (MESH:C085321), SDS (MESH:D012967), PFA (MESH:C003043), TCEP (MESH:C080938), H (MESH:D006859), ACN (MESH:C084683), K (MESH:D011188), Alexa Fluor 488 (MESH:C000711379), IAA (MESH:D007460), Tween 20 (MESH:D011136), cysteine (MESH:D003545), nocodazole (MESH:D015739), methionine (MESH:D008715), M2 (MESH:C034584), DMEM (-), mineral oil (MESH:D008899), DAPI (MESH:C007293), GTP (MESH:D006160), dithiothreitol (MESH:D004229), PVDF (MESH:C024865), FA (MESH:C030544), 3-Isobutyl-1-methylxanthine (MESH:D015056)
- **Species:** Mus musculus (house mouse, species) [taxon 10090], Homo sapiens (human, species) [taxon 9606], Drosophila melanogaster (fruit fly, species) [taxon 7227], Xenopus laevis (African clawed frog, species) [taxon 8355], C.elegans [taxon 328850]
- **Cell lines:** HeLa — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_0030), PAN — Homo sapiens (Human), Chronic myelogenous leukemia, BCR-ABL1 positive, Cancer cell line (CVCL_XR32), HEK293T — Homo sapiens (Human), Transformed cell line (CVCL_0063), C57BL/6J — Mus musculus (Mouse), Transformed cell line (CVCL_C0MW)

## Full text

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## Figures

8 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12311101/full.md

## References

12 references — full list in the complete paper: https://tomesphere.com/paper/PMC12311101/full.md

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Source: https://tomesphere.com/paper/PMC12311101