In vitro reconstitution of motor-driven nuclear oscillations

分子马达在细胞分裂和细胞内运输等基本细胞功能中起着至关重要的作用。虽然我们对单个马达的行为已经了解很多，但对于导致细胞协同运动的多个马达之间的相互作用还没有很好的理解。裂殖酵母裂殖酵母（Schizosophylomycespombe）的减数分裂核振荡是一种研究微管动力蛋白马达驱动的细胞内运动的简单模型过程，本项目将建立一个“人工细胞”系统，首次在体外可控和简化的环境中研究核振荡。我们将开发一种新的检测方法，其中电机可以动态地附着和脱离人工细胞壁，这一过程被定义为对体内振荡至关重要。通过研究这些表面上人工制备的反平行微管对的滑动运动，将确定获得振荡所需的最小组件和条件，并研究各个参数（如马达的结合强度）对振荡的影响。这些研究的结果将阐明导致活细胞大规模运动的马达集体行为的基本原理。

Molecular motors play a crucial role in fundamental cell functions such as cell division and intracellular transport. Although much is known about the behavior of individual motors, the interplay of a multitude of motors, which leads to concerted movements in the cell, is not yet well understood. Meiotic nuclear oscillations in the fission yeast Schizosaccharomyces pombe have been used as an easily accessible model process to study intracellular movements driven by dynein motors operating on microtubules.In this project, an “artificial cell” system will be set up which will for the first time allow for investigating nuclear oscillations in a controlled and simplified environment in vitro. We will develop a novel assay in which motors can dynamically attach and detach from the artificial cell wall, a process defined to be essential for the oscillations in vivo. By studying the gliding motion of artificially prepared, anti-parallel microtubule doublets on these surfaces, the minimal set of components and conditions required to obtain oscillations will be identified, and the impact of individual parameters, such as the binding strength of the motors, on the oscillations will be studied. The results of these investigations will elucidate the fundamentals of the collective behavior of motors leading to large-scale movements in living cells.