Ye Anna A, Maresca Thomas J
Biology Department, University of Massachusetts, Amherst, MA, United States; Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, MA, United States.
Biology Department, University of Massachusetts, Amherst, MA, United States; Molecular and Cellular Biology Graduate Program, University of Massachusetts, Amherst, MA, United States.
Methods Cell Biol. 2018;144:165-184. doi: 10.1016/bs.mcb.2018.03.007. Epub 2018 Apr 10.
Productive chromosome movements require that a large multiprotein complex called the kinetochore assemble on sister centromeres. The kinetochore fulfills two critical functions as (1) the physical linkage between chromosomes and spindle microtubules and (2) a mechanomolecular sensor that relays a spindle assembly checkpoint signal delaying anaphase onset until chromosomes are attached to spindle microtubules and bioriented. Given its central roles in such a vital process, the kinetochore is one of the most important force-transducing structures in cells; yet it has been technically challenging to measure kinetochore forces. Barriers to measuring cellular forces have begun to be broken by the development of fluorescence-based tension sensors. In this chapter, two methods will be described for measuring kinetochore forces in living cells and strategies for applying these sensors to other force-transducing processes and molecules will be discussed.
有效的染色体运动需要一种名为动粒的大型多蛋白复合体在姐妹着丝粒上组装。动粒发挥着两个关键功能：一是作为染色体与纺锤体微管之间的物理连接；二是作为一种机械分子传感器，传递纺锤体组装检查点信号，延迟后期开始，直到染色体附着于纺锤体微管并实现双极定向。鉴于其在这一重要过程中的核心作用，动粒是细胞中最重要的力转导结构之一；然而，测量动粒力在技术上一直具有挑战性。基于荧光的张力传感器的发展已开始打破测量细胞力的障碍。在本章中，将描述两种在活细胞中测量动粒力的方法，并讨论将这些传感器应用于其他力转导过程和分子的策略。
