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Abstract：
The spindle assembly checkpoint (SAC) is critical for sensing defective microtubule-kinetochore attachments and tension across the kinetochore and functions to arrest cells in prometaphase to allow time to repair any errors before proceeding into anaphase. Dysregulation of the SAC leads to chromosome segregation errors that have been linked to human diseases like cancer. Although much has been learned about the composition of the SAC and the factors that regulate its activity, the proximity associations of core SAC components have not been explored in a systematic manner. Here, we have taken a BioID2-proximity-labeling proteomic approach to define the proximity protein environment for each of the five core SAC proteins BUB1, BUB3, BUBR1, MAD1L1, and MAD2L1 in mitotic-enriched populations of cells where the SAC is active. These five protein association maps were integrated to generate a SAC proximity protein network that contains multiple layers of information related to core SAC protein complexes, protein-protein interactions, and proximity associations. Our analysis validated many known SAC complexes and protein-protein interactions. Additionally, it uncovered new protein associations, including the ELYS-MAD1L1 interaction that we have validated, which lend insight into the functioning of core SAC proteins and highlight future areas of investigation to better understand the SAC.
摘要：
纺锤体组装检查点（SAC）对于感测有缺陷的微管-动粒附着和整个动粒的张力至关重要，并且具有阻止前中期细胞的功能，以便在进入后期之前有时间修复任何错误。SAC的失调导致与癌症等人类疾病有关的染色体分离错误。尽管人们对SAC的组成和调节其活性的因素有了很多了解，核心SAC组件的邻近性关联尚未得到系统的探索。这里,我们采用了BioID2-邻近标记蛋白质组学方法,在有丝分裂富集的SAC有活性的细胞群中,确定了5种核心SAC蛋白BUB1,BUB3,BUBR1,MAD1L1和MAD2L1中每一种的邻近蛋白环境.整合这五个蛋白质关联图，以生成SAC邻近蛋白质网络，该网络包含与核心SAC蛋白质复合物相关的多层信息，蛋白质-蛋白质相互作用，和邻近协会。我们的分析验证了许多已知的SAC复合物和蛋白质-蛋白质相互作用。此外,它发现了新的蛋白质关联，包括我们已经验证的ELYS-MAD1L1相互作用，这有助于深入了解核心SAC蛋白的功能，并突出未来的研究领域，以更好地了解SAC。
