Laboratory for Bio- and Nano-Photonics, Department of Microsystems Engineering-IMTEK, University of Freiburg, Freiburg, Germany.
Biophys J. 2013 Sep 3;105(5):1171-81. doi: 10.1016/j.bpj.2013.07.038.
The cytoskeletal protein MreB is an essential component of the bacterial cell-shape generation system. Using a superresolution variant of total internal reflection microscopy with structured illumination, as well as three-dimensional stacks of deconvolved epifluorescence microscopy, we found that inside living Bacillus subtilis cells, MreB forms filamentous structures of variable lengths, typically not longer than 1 μm. These filaments move along their orientation and mainly perpendicular to the long bacterial axis, revealing a maximal velocity at an intermediate length and a decreasing velocity with increasing filament length. Filaments move along straight trajectories but can reverse or alter their direction of propagation. Based on our measurements, we provide a mechanistic model that is consistent with all observations. In this model, MreB filaments mechanically couple several motors that putatively synthesize the cell wall, whereas the filaments' traces mirror the trajectories of the motors. On the basis of our mechanistic model, we developed a mathematical model that can explain the nonlinear velocity length dependence. We deduce that the coupling of cell wall synthesis motors determines the MreB filament transport velocity, and the filament mechanically controls a concerted synthesis of parallel peptidoglycan strands to improve cell wall stability.
细胞骨架蛋白 MreB 是细菌细胞形状生成系统的重要组成部分。我们使用带有结构照明的超分辨率全内反射显微镜以及经过反卷积的荧光显微镜的三维堆栈,发现活的枯草芽孢杆菌细胞内,MreB 形成长度可变的丝状结构,通常不超过 1 微米。这些纤维沿其方向并主要垂直于长细菌轴移动,在中间长度处显示出最大速度,并随着纤维长度的增加而速度降低。纤维沿直线轨迹移动,但可以反转或改变其传播方向。根据我们的测量结果,我们提供了一个与所有观察结果一致的机械模型。在该模型中,MreB 纤维机械地连接几个假定合成细胞壁的马达,而纤维的轨迹反映了马达的轨迹。基于我们的机械模型,我们开发了一个可以解释非线性速度长度依赖性的数学模型。我们推断出细胞壁合成马达的耦合决定了 MreB 纤维的运输速度,并且纤维机械地控制平行肽聚糖链的协同合成,以提高细胞壁稳定性。