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机械力会对纺锤体微管晶格造成局部损伤、重塑并使其稳定。

Mechanical force locally damages, remodels and stabilizes the lattice of spindle microtubules.

作者信息

Rux Caleb J, Chong Megan K, Myers Sean, Cho Nathan H, Dumont Sophie

机构信息

Department of Bioengineering & Therapeutic Sciences, San Francisco, CA 94158, USA.

UC Berkeley-UCSF Graduate Program in Bioengineering, San Francisco, CA 94158, USA.

出版信息

bioRxiv. 2025 Jun 6:2025.06.05.657915. doi: 10.1101/2025.06.05.657915.

DOI:10.1101/2025.06.05.657915
PMID:40501666
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12157524/
Abstract

To segregate chromosomes at cell division, the spindle must maintain its structure under force. How it does so remains poorly understood. To address this question, we use microneedle manipulation to apply local force to spindle microtubule bundles, kinetochore-fibers (k-fibers), inside mammalian cells. We show that local load directly fractures k-fibers, and that newly created plus-ends often have arrested dynamics, resisting depolymerization. Force alone, without fracture, is sufficient for spindle microtubule stabilization, as revealed by laser ablating k-fibers under local needle force. Doublecortin, which binds a compacted microtubule lattice, is lost around the force application site, suggesting local force-induced structural remodeling. In turn, EB1, which recognizes GTP-tubulin, is locally enriched at stabilization sites, both before and after force-induced fracture. Together, our findings support a model where force-induced damage leads to local spindle microtubule lattice remodeling and stabilization, which we propose reinforces the spindle where it experiences critical loads.

摘要

为了在细胞分裂时分离染色体,纺锤体必须在受力情况下维持其结构。其具体机制仍知之甚少。为了解决这个问题,我们使用微针操作对哺乳动物细胞内的纺锤体微管束,即动粒微管(k纤维)施加局部力。我们发现局部负载会直接使k纤维断裂,并且新产生的正端通常具有停滞的动力学,抵抗解聚。如在局部针力作用下激光消融k纤维所显示的,仅有力而无断裂就足以使纺锤体微管稳定。与紧密微管晶格结合的双皮质素在力施加部位周围消失,表明局部力诱导了结构重塑。反过来,识别GTP微管蛋白的EB1在力诱导断裂前后均在稳定部位局部富集。总之,我们的研究结果支持一种模型,即力诱导的损伤导致局部纺锤体微管晶格重塑和稳定,我们认为这加强了纺锤体承受关键负载的部位。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a997/12157524/fe24781433dd/nihpp-2025.06.05.657915v1-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a997/12157524/efc001b1edd8/nihpp-2025.06.05.657915v1-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a997/12157524/fe24781433dd/nihpp-2025.06.05.657915v1-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a997/12157524/efc001b1edd8/nihpp-2025.06.05.657915v1-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a997/12157524/fe24781433dd/nihpp-2025.06.05.657915v1-f0006.jpg

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本文引用的文献

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J Cell Biol. 2025 Oct 6;224(10). doi: 10.1083/jcb.202501070. Epub 2025 Aug 11.
2
The GTP-tubulin cap is not the determinant of microtubule end stability in cells.GTP-微管蛋白帽并不是细胞中微管末端稳定性的决定因素。
Mol Biol Cell. 2024 Oct 1;35(10):br19. doi: 10.1091/mbc.E24-07-0307. Epub 2024 Sep 11.
3
Kinetochore-fiber lengths are maintained locally but coordinated globally by poles in the mammalian spindle.
着丝点微管的长度在局部得到维持,但在哺乳动物纺锤体中,通过两极进行全局协调。
Elife. 2023 Jul 3;12:e85208. doi: 10.7554/eLife.85208.
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Compressive forces stabilize microtubules in living cells.压缩力使活细胞中的微管稳定。
Nat Mater. 2023 Jul;22(7):913-924. doi: 10.1038/s41563-023-01578-1. Epub 2023 Jun 29.
5
CLASPs stabilize the pre-catastrophe intermediate state between microtubule growth and shrinkage.衔接蛋白稳定微管生长和收缩之间的灾变前中间状态。
J Cell Biol. 2023 Jul 3;222(7). doi: 10.1083/jcb.202107027. Epub 2023 May 15.
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Mechanisms underlying spindle assembly and robustness.纺锤体组装和稳定性的机制。
Nat Rev Mol Cell Biol. 2023 Aug;24(8):523-542. doi: 10.1038/s41580-023-00584-0. Epub 2023 Mar 28.
7
CSPP1 stabilizes growing microtubule ends and damaged lattices from the luminal side.CSPP1 从腔侧稳定生长的微管末端和受损的晶格。
J Cell Biol. 2023 Apr 3;222(4). doi: 10.1083/jcb.202208062. Epub 2023 Feb 8.
8
Modeling and mechanical perturbations reveal how spatially regulated anchorage gives rise to spatially distinct mechanics across the mammalian spindle.建模和力学干扰揭示了空间调节的附着是如何在哺乳动物纺锤体中产生空间上不同的力学特性的。
Elife. 2022 Nov 8;11:e79558. doi: 10.7554/eLife.79558.
9
Evidence for a HURP/EB free mixed-nucleotide zone in kinetochore-microtubules.有证据表明动粒微管中存在 HURP/EB 游离混合核苷酸区。
Nat Commun. 2022 Aug 10;13(1):4704. doi: 10.1038/s41467-022-32421-x.
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A kinesin-1 variant reveals motor-induced microtubule damage in cells.一种肌球蛋白-1 变体揭示了马达诱导的细胞中微管损伤。
Curr Biol. 2022 Jun 6;32(11):2416-2429.e6. doi: 10.1016/j.cub.2022.04.020. Epub 2022 May 2.