CENP-E as an essential component of the mitotic checkpoint in vitro
CENP-E as an essential component of the mitotic checkpoint in vitro
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DOI：
10.1016/s0092-8674(00)00070-2
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发表时间：
2000-09-15
期刊：
影响因子：
64.5
通讯作者：
Cleveland, DW
中科院分区：
文献类型：
--
作者：
Abrieu, A;Kahana, JA;Cleveland, DW
准确的染色单体分离由检查点机制监测，该机制延迟后期开始，直到所有着丝粒正确地附着在有丝分裂纺锤体上。使用爪蟾卵提取物，现在发现着丝点相关的微管马达蛋白CENP-E是建立和维持这个检查点所必需的。当CENP-E功能被免疫消耗或抗体添加破坏时，提取物不能抑制纺锤体损伤。有丝分裂阻滞可以通过添加高水平的可溶性MAD2来恢复，这表明CENP-E的缺失消除了着丝酶依赖的信号传导，但没有消除检查点信号转导的下游步骤。因为它直接结合纺锤体微管和着丝点相关的检查点激酶BUBR1，所以CENP-E是脊椎动物检查点的中心成分，以微管依赖的方式调节信号活性。
Accurate chromatid separation is monitored by a checkpoint mechanism that delays anaphase onset until all centromeres are correctly attached to the mitotic spindle. Using Xenopus egg extracts, the kinetochore-associated microtubule motor protein CENP-E is now found to be required for establishing and maintaining this checkpoint. When CENP-E function is disrupted by immunodepletion or antibody addition, extracts fail to arrest in response to spindle damage. Mitotic arrest can be restored by addition of high levels of soluble MAD2, demonstrating that the absence of CENP-E eliminates kinetochore-dependent signaling but not the downstream steps in checkpoint signal transduction. Because it directly binds both to spindle microtubules and to the kinetochore-associated checkpoint kinase BUBR1, CENP-E is a central component in the vertebrate checkpoint that modulates signaling activity in a microtubule-dependent manner.