Kent Ian A, Rane Parag S, Dickinson Richard B, Ladd Anthony J C, Lele Tanmay P
Department of Chemical Engineering, University of Florida, Gainesville, FL, United States of America.
PLoS One. 2016 Mar 14;11(3):e0151322. doi: 10.1371/journal.pone.0151322. eCollection 2016.
Microtubules have a persistence length of the order of millimeters in vitro, but inside cells they bend over length scales of microns. It has been proposed that polymerization forces bend microtubules in the vicinity of the cell boundary or other obstacles, yet bends develop even when microtubules are polymerizing freely, unaffected by obstacles and cell boundaries. How these bends are formed remains unclear. By tracking the motions of microtubules marked by photobleaching, we found that in LLC-PK1 epithelial cells local bends develop primarily by plus-end directed transport of portions of the microtubule contour towards stationary locations (termed pinning points) along the length of the microtubule. The pinning points were transient in nature, and their eventual release allowed the bends to relax. The directionality of the transport as well as the overall incidence of local bends decreased when dynein was inhibited, while myosin inhibition had no observable effect. This suggests that dynein generates a tangential force that bends microtubules against stationary pinning points. Simulations of microtubule motion and polymerization accounting for filament mechanics and dynein forces predict the development of bends of size and shape similar to those observed in cells. Furthermore, simulations show that dynein-generated bends at a pinning point near the plus end can cause a persistent rotation of the tip consistent with the observation that bend formation near the tip can change the direction of microtubule growth. Collectively, these results suggest a simple physical mechanism for the bending of growing microtubules by dynein forces accumulating at pinning points.
微管在体外的持久长度约为毫米级,但在细胞内它们会在微米级的长度尺度上弯曲。有人提出,聚合力会使微管在细胞边界或其他障碍物附近弯曲,然而,即使微管在自由聚合时不受障碍物和细胞边界的影响,弯曲也会出现。这些弯曲是如何形成的仍不清楚。通过追踪光漂白标记的微管运动,我们发现,在LLC-PK1上皮细胞中,局部弯曲主要是通过微管轮廓的部分向微管长度方向上的固定位置(称为固定点)进行正端定向运输而形成的。固定点本质上是短暂的,它们最终的释放使弯曲得以松弛。当动力蛋白被抑制时,运输的方向性以及局部弯曲的总体发生率都会降低,而肌球蛋白抑制则没有可观察到的影响。这表明动力蛋白产生了一种切向力,使微管靠着固定点弯曲。考虑到细丝力学和动力蛋白力的微管运动和聚合模拟预测了与细胞中观察到的大小和形状相似的弯曲的形成。此外,模拟表明,在正端附近的固定点处由动力蛋白产生的弯曲会导致尖端持续旋转,这与尖端附近弯曲形成可改变微管生长方向的观察结果一致。总的来说,这些结果表明了一种简单的物理机制,即通过在固定点积累的动力蛋白力使生长中的微管弯曲。