Structural Cellular Biology Unit, Okinawa Institute of Science and Technology, 904-0495, Okinawa, Japan.
Mechanical Engineering and Microfabrication Support Section, Okinawa Institute of Science and Technology, 904-0495, Okinawa, Japan.
Nat Commun. 2018 Oct 18;9(1):4323. doi: 10.1038/s41467-018-06887-7.
FtsZ is the main regulator of bacterial cell division. It has been implicated in acting as a scaffolding protein for other division proteins, a force generator during constriction, and more recently, as an active regulator of septal cell wall production. FtsZ assembles into a heterogeneous structure coined the Z-ring due to its resemblance to a ring confined by the midcell geometry. Here, to establish a framework for examining geometrical influences on proper Z-ring assembly and dynamics, we sculpted Escherichia coli cells into unnatural shapes using division- and cell wall-specific inhibitors in a micro-fabrication scheme. This approach allowed us to examine FtsZ behavior in engineered Z-squares and Z-hearts. We use stimulated emission depletion (STED) nanoscopy to show that FtsZ clusters in sculpted cells maintain the same dimensions as their wild-type counterparts. Based on our results, we propose that the underlying membrane geometry is not a deciding factor for FtsZ cluster maintenance and dynamics in vivo.
FtsZ 是细菌细胞分裂的主要调节蛋白。它被认为是其他分裂蛋白的支架蛋白，在收缩过程中充当力发生器，最近还被认为是隔膜细胞壁生成的活性调节剂。FtsZ 组装成一种不均匀的结构，称为 Z 环，因为它类似于由中隔几何形状限制的环。在这里，为了建立一个框架来研究几何形状对适当的 Z 环组装和动力学的影响，我们使用了一种微制造方案，使用分裂和细胞壁特异性抑制剂将大肠杆菌细胞塑造成非自然形状。这种方法使我们能够在工程 Z 正方形和 Z 心脏中检查 FtsZ 的行为。我们使用受激发射损耗（STED）纳米显微镜显示，在塑形细胞中，FtsZ 簇保持与野生型细胞相同的尺寸。根据我们的结果，我们提出，潜在的膜几何形状不是体内 FtsZ 簇维持和动力学的决定因素。
