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首页> 外文期刊>Cell cycle >Spindle checkpoint maintenance requires Ame1 and Okp1.
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Spindle checkpoint maintenance requires Ame1 and Okp1.

机译：主轴检查点维护需要Ame1和Okp1。

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Kinetochore proteins are required for high fidelity chromosome segregation and as a platform for checkpoint signaling. Ame1 is an essential component of the COMA (Ctf19, Okp1, Mcm21, Ame1) sub-complex of the central kinetochore of budding yeast. In this study, we describe the isolation and characterization of an Ame1 conditional mutant, ame1-4. ame1-4 cells exhibit chromosome segregation defects and Mad2-dependent cell cycle delay similar to okp1-5 cells. However, the viability of ame1-4 cells is markedly reduced relative to wild type and okp1-5 cells after three hours at restrictive temperature. To determine if ame1-4 cells enter anaphase with mis-segregated chromosomes, we monitored the localization of Bub3:VFP as a marker for anaphase onset. ame1-4 cells containing mis-segregated sister chromatids initially accumulate Bub3:VFP at kinetochores, indicating checkpoint activation and a metaphase arrest. Subsequently, Bub3:VFP de-localizes and cells reinitiate DNA duplication and budding without cytokinesis in the presence of un-segregated chromosomes. Overexpression of OKP1 in ame1-4 cells restores ame1-4 protein localization and a stable arrest. Based on our results, we propose that Ame1 and Okp1 are required for a sustained checkpoint arrest in the presence of mis-segregated chromosomes. Our results suggest that checkpoint response might be controlled not only at the level of activation but also via signals that ensure maintenance of the response.
机译：线粒体蛋白是高保真染色体分离和检查站信号转导的平台所必需的。 Ame1是萌芽酵母中心动粒的COMA（Ctf19，Okp1，Mcm21，Ame1）亚复合物的重要组成部分。在这项研究中，我们描述了Ame1条件突变蛋白ame1-4的分离和表征。 ame1-4细胞与okp1-5细胞相似，表现出染色体分离缺陷和Mad2依赖性细胞周期延迟。然而，在限制性温度下三小时后，相对于野生型和okp1-5细胞，ame1-4细胞的活力明显降低。为了确定ame1-4细胞是否进入染色体分离错误的后期，我们监测了Bub3：VFP的定位，作为后期发病的标记。包含错误分离的姐妹染色单体的ame1-4细胞最初在动植物体内积累Bub3：VFP，表明检查点激活和中期停滞。随后，在存在未分离的染色体的情况下，Bub3：VFP脱位，细胞重新启动DNA复制和出芽，而没有胞质分裂。 OKP1在ame1-4细胞中的过表达恢复了ame1-4蛋白的定位和稳定的阻滞。根据我们的结果，我们建议在存在错误分离的染色体的情况下，Ame1和Okp1对于持续的检查点逮捕是必需的。我们的结果表明，检查点响应不仅可以在激活级别进行控制，还可以通过确保响应保持的信号进行控制。

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