Department of Cell & Molecular Biology, The University of Rhode Island, Kingston, Rhode Island, USA.
Department of Cell & Molecular Biology, The University of Rhode Island, Kingston, Rhode Island, USA.
J Biol Chem. 2021 Jan-Jun;296:100162. doi: 10.1074/jbc.RA120.013866. Epub 2020 Dec 10.
MinD is a cell division ATPase in Escherichia coli that oscillates from pole to pole and regulates the spatial position of the cell division machinery. Together with MinC and MinE, the Min system restricts assembly of the FtsZ-ring to midcell, oscillating between the opposite ends of the cell and preventing FtsZ-ring misassembly at the poles. Here, we show that the ATP-dependent bacterial proteasome complex ClpXP degrades MinD in reconstituted degradation reactions in vitro and in vivo through direct recognition of the MinD N-terminal region. MinD degradation is enhanced during stationary phase, suggesting that ClpXP regulates levels of MinD in cells that are not actively dividing. ClpXP is a major regulator of growth phase-dependent proteins, and these results suggest that MinD levels are also controlled during stationary phase. In vitro, MinC and MinD are known to coassemble into linear polymers; therefore, we monitored copolymers assembled in vitro after incubation with ClpXP and observed that ClpXP promotes rapid MinCD copolymer destabilization and direct MinD degradation by ClpXP. The N terminus of MinD, including residue Arg 3, which is near the ATP-binding site in sequence, is critical for degradation by ClpXP. Together, these results demonstrate that ClpXP degradation modifies conformational assemblies of MinD in vitro and depresses Min function in vivo during periods of reduced proliferation.
MinD 是大肠杆菌中的一种细胞分裂 ATP 酶,它从极到极来回振荡,调节细胞分裂机制的空间位置。与 MinC 和 MinE 一起,Min 系统将 FtsZ 环的组装限制在细胞中部,在细胞的两端之间振荡,防止 FtsZ 环在极处错误组装。在这里,我们表明,依赖于 ATP 的细菌蛋白酶体复合物 ClpXP 在体外重构的降解反应中和体内通过直接识别 MinD N 端区域来降解 MinD。在静止期,MinD 的降解增强,这表明 ClpXP 调节不活跃分裂的细胞中 MinD 的水平。ClpXP 是生长阶段依赖性蛋白的主要调节剂,这些结果表明 MinD 水平也在静止期受到控制。在体外,已知 MinC 和 MinD 会共同组装成线性聚合物;因此,我们在与 ClpXP 孵育后监测体外组装的共聚物,并观察到 ClpXP 促进 MinCD 共聚物的快速不稳定,并直接由 ClpXP 降解 MinD。MinD 的 N 端,包括序列中靠近 ATP 结合位点的残基 Arg3,对于 ClpXP 的降解至关重要。总之,这些结果表明,ClpXP 降解在体外改变了 MinD 的构象组装,并在增殖减少期间在体内抑制 Min 功能。