Zheng Ge, Ouyang Zhuqing, Yu Hongtao
Howard Hughes Medical Institute, Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX, 75390, USA.
Methods Mol Biol. 2017;1515:37-53. doi: 10.1007/978-1-4939-6545-8_3.
During the cell cycle, duplicated sister chromatids become physically connected during S phase through a process called sister-chromatid cohesion. Cohesion is terminated during the metaphase-to-anaphase transition to trigger sister-chromatid segregation. The establishment and dissolution of cohesion are highly regulated by the cohesin complex and its multitude of regulators. In particular, the cohesin regulator Wapl promotes the release of cohesin from chromosomes during both interphase and mitosis. Here, we describe in vitro protein binding assays between Wapl and a cohesin subcomplex, and cellular assays in human cells that probe the functions of Wapl in cohesin release.
在细胞周期中，复制后的姐妹染色单体在S期通过一个称为姐妹染色单体黏连的过程实现物理连接。黏连在中期到后期的转变过程中终止，从而触发姐妹染色单体的分离。黏连的建立和解除受到黏连蛋白复合体及其众多调节因子的高度调控。特别是，黏连蛋白调节因子Wapl在间期和有丝分裂期间都能促进黏连蛋白从染色体上释放。在这里，我们描述了Wapl与黏连蛋白亚复合体之间的体外蛋白质结合试验，以及在人类细胞中探究Wapl在黏连蛋白释放中的功能的细胞试验。
