RZZ and Mad1 dynamics in Drosophila mitosis.

RZZ and Mad1 dynamics in Drosophila mitosis.
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DOI:
10.1007/s10577-015-9472-x
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发表时间:
2015-06
影响因子:
2.6
通讯作者:
Karess, Roger E.
Karess, Roger E.
中科院分区:
生物学2区
文献类型:
--
作者:
Defachelles, Lenaig;Raich, Natacha;Terracol, Regine;Baudin, Xavier;Williams, Byron;Goldberg, Michael;Karess, Roger E.

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着丝粒上 Mad1 的存在与否是纺锤体组装检查点 (SAC) 活性的主要决定因素,SAC 是一种监视机制,如果一个或多个着丝粒未附着在纺锤体纤维上,则会延迟后期的发生。 Rod、Zw10 和 Zwilch (RZZ) 复合体是调节着丝粒 Mad1 水平的因素之一,它是 Mad1 通过尚不清楚的机制募集所必需的。尽管有报道 Mad1 被稳定地募集到独立的着丝粒上,但 Mad1 和 RZZ 在着丝粒上的相对动力学和相互作用尚未得到广泛研究。在这项研究中,我们直接比较了未附着的果蝇着丝粒上 Mad1-绿色荧光蛋白 (GFP) 的周转动态与用 GFP-Rod 或 GFP-Zw10 标记的 RZZ 的周转动态。我们发现近 40% 的着丝粒结合 Mad1 具有显着的动态成分,其半衰期为 12 秒。相比之下,RZZ 在未附着的动粒上基本上是稳定的。此外,我们报告说,一小部分 RZZ 和 Mad1 可以共免疫沉淀,这表明遗传决定的招募层次(其中 Mad1 依赖于 RZZ)可能反映了两种复合物的物理关联。本文的在线版本 (doi:10.1007/s10577-015-9472-x) 包含补充材料,可供授权用户使用。
The presence or absence of Mad1 at kinetochores is a major determinant of spindle assembly checkpoint (SAC) activity, the surveillance mechanism that delays anaphase onset if one or more kinetochores remain unattached to spindle fibers. Among the factors regulating the levels of Mad1 at kinetochores is the Rod, Zw10, and Zwilch (RZZ) complex, which is required for Mad1 recruitment through a mechanism that remains unknown. The relative dynamics and interactions of Mad1 and RZZ at kinetochores have not been extensively investigated, although Mad1 has been reported to be stably recruited to unattached kinetochores. In this study, we directly compare Mad1-green fluorescent protein (GFP) turnover dynamics on unattached Drosophila kinetochores with that of RZZ, tagged either with GFP-Rod or GFP-Zw10. We find that nearly 40 % of kinetochore-bound Mad1 has a significant dynamic component, turning over with a half-life of 12 s. RZZ in contrast is essentially stable on unattached kinetochores. In addition, we report that a fraction of RZZ and Mad1 can co-immunoprecipitate, indicating that the genetically determined recruitment hierarchy (in which Mad1 depends on RZZ) may reflect a physical association of the two complexes. The online version of this article (doi:10.1007/s10577-015-9472-x) contains supplementary material, which is available to authorized users.
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