IDENTIFICATION OF PHOSPHORYLATION SITES AURORA B

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。 在每一次细胞分裂过程中，复制的基因组必须准确地分配给子细胞。细胞使用多种机制来确保染色体通过基于微管的有丝分裂纺锤体高保真地分离。一个关键的检查点涉及Aurora B激酶，它破坏了没有处于张力状态的有缺陷的动粒-微管连接的稳定性。极光B是染色体乘客复合体(CPC)的一员，CPC是一组相互作用的蛋白质，它们通过中期定位到染色体着丝粒区域，然后在后期重新定位到中心纺锤体。我们正在研究在萌芽酵母中通过翻译后修饰对CPC的调控。我们将在体外通过候选激酶磷酸化的纯化CPC的质谱学来鉴定翻译后修饰。到目前为止，我们已经在Aurora B的激活剂INCENP上确定了一些潜在的自我调节磷酸化位点。未来的工作将探索这些位点的功能，并确定其他调节激酶的修饰，如Mps1和CDK1。这种方法将深入了解细胞分裂过程中运行的局部信号机制，以确保细胞中所有染色体的协调排列和分离。

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. During every cell division, the replicated genome must be distributed accurately to daughter cells. Cells use multiple mechanisms to ensure that chromosomes are segregated with high fidelity by the microtubule-based mitotic spindle. One key checkpoint involves the kinase Aurora B, which destabilizes defective kinetochore-microtubule attachments that are not under tension. Aurora B is a member of the chromosomal passenger complex (CPC), a set of interacting proteins named for their localization to the centromere region of chromosomes through metaphase and subsequent relocalization to the central spindle at anaphase. We are studying the regulation of the CPC by post-translation modification in the budding yeast Saccharomyces cerevisiae. We will identify of post-translational modifications by mass spectroscopy of purified CPC phosphorylated by candidate kinases in vitro. Thus far we have identified a number of potential self-regulatory phosphorylation sites on the activator of Aurora B, INCENP. Future work will explore the function of these sites and identify modifications by other regulatory kinases such as Mps1 and Cdk1. This approach will give insight into the local signaling mechanisms that operate during cell division to ensure the coordinated alignment and segregation of all of the chromosomes in a cell.