光漂白动态坐标系中马达驱动的微管扩散

Soichi Hirokawa, Heun Jin Lee, Rachel A Banks, Ana I Duarte, Bibi Najma, Matt Thomson, Rob Phillips
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引用次数: 0

摘要

在细胞系统中,马达驱动的细胞骨架重塑往往伴随着局部尺度上的类似扩散效应,但是很难将有序化过程(如网络的主动收缩)与这种主动扩散区分开来。利用可光二聚化的驱动蛋白马达在空间上控制微管网络的形成和收缩,我们特意将网格图案光漂白到长丝网络上,作为一个瞬时动态坐标系来观察微管剩余荧光方格的变形和平移。我们发现,微管网络以马达速度设定的速率收缩,但在整个收缩网络中伴随着类似扩散的扩散,其有效扩散常数比自由扩散的微管低两个数量级。我们进一步发现,在微米尺度上,无论条件如何,扩散时间尺度仅比平流时间尺度慢约 3 倍,这表明这种扩散行为的全局收缩和长时间松弛都是由马达驱动的,但在网络主体内表现出局部竞争。
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Motor-driven microtubule diffusion in a photobleached dynamical coordinate system
Motor-driven cytoskeletal remodeling in cellular systems can often be accompanied by a diffusive-like effect at local scales, but distinguishing the contributions of the ordering process, such as active contraction of a network, from this active diffusion is difficult to achieve. Using light-dimerizable kinesin motors to spatially control the formation and contraction of a microtubule network, we deliberately photobleach a grid pattern onto the filament network serving as a transient and dynamic coordinate system to observe the deformation and translation of the remaining fluorescent squares of microtubules. We find that the network contracts at a rate set by motor speed but is accompanied by a diffusive-like spread throughout the bulk of the contracting network with effective diffusion constant two orders of magnitude lower than that for a freely-diffusing microtubule. We further find that on micron scales, the diffusive timescale is only a factor of approximately 3 slower than that of advection regardless of conditions, showing that the global contraction and long-time relaxation from this diffusive behavior are both motor-driven but exhibit local competition within the network bulk.
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