KINASE REGULATION DURING THE METAPHASE-ANAPHASE TRANSITION IN MITOSIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 忠实的染色体分离依赖于监测微管-动粒附着的细胞机制。正确的附着允许细胞进入下一个细胞周期，而错误会导致细胞周期停滞，直到它们被纠正。触发这种停滞的信号要么是动粒和微管之间缺乏连接，要么是纺锤体缺乏张力。已知这一信号涉及纺锤体检查点蛋白和其他有丝分裂酶和磷酸酶。然而，这些组分如何协调作用来调节细胞周期还没有完全了解，部分原因是许多有丝分裂酶在整个细胞周期中执行多种功能。为了解决这个问题，我们正在对来自不同细胞周期阶段和微管-动粒附着的不同状态的有丝分裂酶复合体进行亲和纯化和质谱分析，以确定蛋白质-蛋白质相互作用和翻译后修饰的任何差异。这将使我们能够理解这些复合体是如何调节的，以响应微管-动粒连接的错误。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Faithful chromosome segregation relies on the cellular mechanism that monitors microtubule-kinetochore attachment. The correct attachment allows cells to progress to the next cell cycle, while errors cause cell cycle arrest until they become corrected. The signal to trigger this arrest is either lack of attachment between kinetochore and microtubule or lack of tension in the spindle. This signaling is known to involve spindle checkpoint proteins and other mitotic kinases and phosphatases. However, how these components function coordinately to regulate the cell cycle has not been fully understood, partly because many mitotic kinases conduct multiple functions throughout the cell cycle. To address this issue, we are conducting affinity purification and mass spectrometric analysis of mitotic kinase complexes from different cells cycle stages and in different states of microtubule-kinetochore attachment to identify any differences in protein-protein interactions and post-translational modifications. This will allow us to understand how these complexes are regulated in response to errors in microtubule-kinetochore attachment.