Klebig Christiane, Toso Alberto, Borusu Satyarebala, Meraldi Patrick
Institute of Biochemistry, ETH Zurich, Zurich, Switzerland.
Methods Mol Biol. 2009;545:205-20. doi: 10.1007/978-1-60327-993-2_12.
During cell division microtubules of the mitotic spindle segregate the duplicated chromosomes into the two daughter cells. Chromosome-microtubule attachment is mediated by kinetochores, multiprotein complexes assembled on specialized regions of the DNA. Kinetochores modulate microtubule dynamics to generate the forces necessary to power chromosome movement and regulate the spindle checkpoint. Errors in kinetochore function can cause aneuploidy, a hallmark of 80% of solid tumors in humans, suggesting a fundamental link to tumorigenesis. Human kinetochores are complex protein machines with over 100 different proteins. Here we present fixed- and live-cell-based assays used to functionally categorize kinetochore proteins with regard to spindle checkpoint activity and kinetochore-microtubule attachment.
在细胞分裂过程中，有丝分裂纺锤体的微管将复制后的染色体分离到两个子细胞中。染色体与微管的附着由动粒介导，动粒是在DNA特定区域组装的多蛋白复合体。动粒调节微管动力学，以产生驱动染色体移动所需的力，并调节纺锤体检查点。动粒功能异常可导致非整倍体，这是80%人类实体瘤的一个标志，提示其与肿瘤发生存在根本联系。人类动粒是由100多种不同蛋白质组成的复杂蛋白质机器。在此，我们展示了基于固定细胞和活细胞的检测方法，用于根据纺锤体检查点活性和动粒与微管的附着对动粒蛋白进行功能分类。
