Zhang Huaiying, Aonbangkhen Chanat, Tarasovetc Ekaterina V, Ballister Edward R, Chenoweth David M, Lampson Michael A
Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Department of Chemistry, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Nat Chem Biol. 2017 Oct;13(10):1096-1101. doi: 10.1038/nchembio.2456. Epub 2017 Aug 14.
Kinetochores act as hubs for multiple activities during cell division, including microtubule interactions and spindle checkpoint signaling. Each kinetochore can act autonomously, and activities change rapidly as proteins are recruited to, or removed from, kinetochores. Understanding this dynamic system requires tools that can manipulate kinetochores on biologically relevant temporal and spatial scales. Optogenetic approaches have the potential to provide temporal and spatial control with molecular specificity. Here we report new chemical inducers of protein dimerization that allow us to both recruit proteins to and release them from kinetochores using light. We use these dimerizers to manipulate checkpoint signaling and molecular motor activity. Our findings demonstrate specialized properties of the CENP-E (kinesin-7) motor for directional chromosome transport to the spindle equator and for maintenance of metaphase alignment. This work establishes a foundation for optogenetic control of kinetochore function, which is broadly applicable to experimental probing of other dynamic cellular processes.
动粒在细胞分裂过程中作为多种活动的枢纽，包括微管相互作用和纺锤体检查点信号传导。每个动粒都能自主发挥作用，并且随着蛋白质被招募到动粒或从动粒上移除，其活动会迅速变化。要理解这个动态系统，需要能够在生物学相关的时间和空间尺度上操纵动粒的工具。光遗传学方法有潜力提供具有分子特异性的时间和空间控制。在此，我们报告了新的蛋白质二聚化化学诱导剂，它们使我们能够利用光将蛋白质招募到动粒上以及从动粒上释放。我们使用这些二聚化剂来操纵检查点信号传导和分子马达活性。我们的研究结果证明了驱动蛋白7（CENP-E）马达在将染色体定向运输到纺锤体赤道以及维持中期排列方面的特殊性质。这项工作为动粒功能的光遗传学控制奠定了基础，这广泛适用于对其他动态细胞过程的实验探究。
