Kinesin-12 KLP-18 contributes to the kinetochore-microtubule poleward flux during the metaphase of C. elegans one-cell embryo
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| Format: | Preprint |
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2025
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| _version_ | 1866909681332518912 |
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| author | Soler, Nina Da Silva, Mathis Tascon, Christophe Chesneau, Laurent Foliard, Pauline Bouvrais, Hélène Pastezeur, Sylvain Marrec, Loïc Le Pecreaux, Jacques |
| author_facet | Soler, Nina Da Silva, Mathis Tascon, Christophe Chesneau, Laurent Foliard, Pauline Bouvrais, Hélène Pastezeur, Sylvain Marrec, Loïc Le Pecreaux, Jacques |
| contents | The mitotic spindle partitions chromosomes during cell division by connecting the poles to kinetochores through microtubules (MTs). Their plus-ends, facing the chromosomes, exhibit dynamic instability, which is critical for proper attachment. The poleward flux implicates the displacement of Mts towards the spindle poles, while plus-ends polymerise. It may result from minus-end depolymerisation (treadmilling), sliding by kinesins (e.g., Kinesin-5), or pushing by chromokinesins. Intriguingly, such flux had not been reported in the C. elegans zygote, despite homologs of flux-associated proteins being present.
To investigate this, we fluorescently labelled Mts and used photobleaching. We observed no global flux; instead, the bleached zone's edges moved inward. The centrosome-facing front reflected MT dynamic instability, but the chromosome-facing front showed faster recovery, suggesting an additional mechanism. This extra velocity was spatially restricted to the vicinity of chromosomes, suggesting that only the kinetochore Mts may undergo flux. Supporting this, flux required key kinetochore regulators: NDC-80, $\text{CLS-2}^\text{CLASP}$, and $\text{ZYG-9}^\text{XMAP215}$. Flux declined as metaphase progressed, correlating with the attachment maturation from lateral to end-on, and was suppressed by SKA-1 recruitment. Classic treadmilling was unlikely, as most kinetochore MTs in C. $elegans$ do not reach spindle poles. Instead, depleting $\text{KLP-18}^\text{KIF15}$, a kinesin that cross-links and organises Mts during meiosis, reduced front movement. We propose that only kinetochore Mts undergo flux, sliding along the spindle Mts, likely powered by KLP-18. This localised sliding contrasts with global flux seen in other systems, and aligns with observations in human cells showing flux reduction as chromosome-to-pole distance increases. |
| format | Preprint |
| id |
arxiv_https___arxiv_org_abs_2507_06620 |
| institution | arXiv |
| publishDate | 2025 |
| record_format | arxiv |
| spellingShingle | Kinesin-12 KLP-18 contributes to the kinetochore-microtubule poleward flux during the metaphase of C. elegans one-cell embryo Soler, Nina Da Silva, Mathis Tascon, Christophe Chesneau, Laurent Foliard, Pauline Bouvrais, Hélène Pastezeur, Sylvain Marrec, Loïc Le Pecreaux, Jacques Subcellular Processes The mitotic spindle partitions chromosomes during cell division by connecting the poles to kinetochores through microtubules (MTs). Their plus-ends, facing the chromosomes, exhibit dynamic instability, which is critical for proper attachment. The poleward flux implicates the displacement of Mts towards the spindle poles, while plus-ends polymerise. It may result from minus-end depolymerisation (treadmilling), sliding by kinesins (e.g., Kinesin-5), or pushing by chromokinesins. Intriguingly, such flux had not been reported in the C. elegans zygote, despite homologs of flux-associated proteins being present. To investigate this, we fluorescently labelled Mts and used photobleaching. We observed no global flux; instead, the bleached zone's edges moved inward. The centrosome-facing front reflected MT dynamic instability, but the chromosome-facing front showed faster recovery, suggesting an additional mechanism. This extra velocity was spatially restricted to the vicinity of chromosomes, suggesting that only the kinetochore Mts may undergo flux. Supporting this, flux required key kinetochore regulators: NDC-80, $\text{CLS-2}^\text{CLASP}$, and $\text{ZYG-9}^\text{XMAP215}$. Flux declined as metaphase progressed, correlating with the attachment maturation from lateral to end-on, and was suppressed by SKA-1 recruitment. Classic treadmilling was unlikely, as most kinetochore MTs in C. $elegans$ do not reach spindle poles. Instead, depleting $\text{KLP-18}^\text{KIF15}$, a kinesin that cross-links and organises Mts during meiosis, reduced front movement. We propose that only kinetochore Mts undergo flux, sliding along the spindle Mts, likely powered by KLP-18. This localised sliding contrasts with global flux seen in other systems, and aligns with observations in human cells showing flux reduction as chromosome-to-pole distance increases. |
| title | Kinesin-12 KLP-18 contributes to the kinetochore-microtubule poleward flux during the metaphase of C. elegans one-cell embryo |
| topic | Subcellular Processes |
| url | https://arxiv.org/abs/2507.06620 |