Regulation of outer kinetochore assembly during meiosis I and II by CENP-A and KNL-2/M18BP1 in C. elegans oocytes.

IF 8.1 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY Current Biology Pub Date : 2024-11-04 Epub Date: 2024-09-30 DOI:10.1016/j.cub.2024.09.004
Laura Bellutti, Nicolas Macaisne, Layla El Mossadeq, Thadshagine Ganeswaran, Julie C Canman, Julien Dumont
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Abstract

During cell division, chromosomes build kinetochores that attach to spindle microtubules. Kinetochores usually form at the centromeres, which contain CENP-A nucleosomes. The outer kinetochore, which is the core attachment site for microtubules, is composed of the KMN network (Knl1c, Mis12c, and Ndc80c complexes) and is recruited downstream of CENP-A and its partner CENP-C. In C. elegans oocytes, kinetochores have been suggested to form independently of CENP-A nucleosomes. Yet kinetochore formation requires CENP-C, which acts in parallel to the nucleoporin MEL-28ELYS. Here, we used a combination of RNAi and Degron-based depletion of CENP-A (or downstream CENP-C) to demonstrate that both proteins are in fact responsible for a portion of outer kinetochore assembly during meiosis I and are essential for accurate chromosome segregation. The remaining part requires the coordinated action of KNL-2 (ortholog of human M18BP1) and of the nucleoporin MEL-28ELYS. Accordingly, co-depletion of CENP-A (or CENP-C) and KNL-2M18BP1 (or MEL-28ELYS) prevented outer kinetochore assembly in oocytes during meiosis I. We further found that KNL-2M18BP1 and MEL-28ELYS are interdependent for kinetochore localization. Using engineered mutants, we demonstrated that KNL-2M18BP1 recruits MEL-28ELYS at meiotic kinetochores through a specific N-terminal domain, independently of its canonical CENP-A loading factor activity. Finally, we found that meiosis II outer kinetochore assembly was solely dependent on the canonical CENP-A/CENP-C pathway. Thus, like in most cells, outer kinetochore assembly in C. elegans oocytes depends on centromeric chromatin. However, during meiosis I, an additional KNL-2M18BP1 and MEL-28ELYS pathway acts in a non-redundant manner and in parallel to canonical centromeric chromatin.

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优雅小鼠卵母细胞减数分裂 I 和 II 期间 CENP-A 和 KNL-2/M18BP1 对外动点组装的调控。
在细胞分裂过程中,染色体会形成连接到纺锤体微管上的动点。动点通常在中心粒上形成,中心粒上含有 CENP-A 核小体。外侧动核是微管的核心附着点,由 KMN 网络(Knl1c、Mis12c 和 Ndc80c 复合物)组成,并在 CENP-A 及其伙伴 CENP-C 的下游被招募。在秀丽隐杆线虫卵母细胞中,有人认为动核的形成与 CENP-A 核小体无关。然而,动芯的形成需要 CENP-C,它与核小体 MEL-28ELYS 平行作用。在这里,我们利用 RNAi 和基于 Degron 的 CENP-A(或下游 CENP-C)耗竭相结合的方法证明,这两种蛋白实际上都负责减数分裂 I 期间外动点心组装的一部分,并且对染色体的准确分离至关重要。其余部分则需要 KNL-2(人类 M18BP1 的直系同源物)和核多聚蛋白 MEL-28ELYS 的协调作用。因此,共同缺失 CENP-A(或 CENP-C)和 KNL-2M18BP1(或 MEL-28ELYS)会阻止减数分裂 I 期卵母细胞中外动核的组装。我们利用工程突变体证明,KNL-2M18BP1 通过一个特异的 N 端结构域将 MEL-28ELYS 募集到减数分裂动点上,而不依赖于其典型的 CENP-A 负载因子活性。最后,我们发现减数分裂 II 的外侧着丝点组装完全依赖于典型的 CENP-A/CENP-C 途径。因此,与大多数细胞一样,优雅小鼠卵母细胞中外动核的组装也依赖于中心染色质。然而,在减数分裂 I 期间,另有一条 KNL-2M18BP1 和 MEL-28ELYS 途径以一种非冗余的方式与典型的中心染色质并行发挥作用。
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来源期刊
Current Biology
Current Biology 生物-生化与分子生物学
CiteScore
11.80
自引率
2.20%
发文量
869
审稿时长
46 days
期刊介绍: Current Biology is a comprehensive journal that showcases original research in various disciplines of biology. It provides a platform for scientists to disseminate their groundbreaking findings and promotes interdisciplinary communication. The journal publishes articles of general interest, encompassing diverse fields of biology. Moreover, it offers accessible editorial pieces that are specifically designed to enlighten non-specialist readers.
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