Department of Molecular Biophysics and Biochemistry, Nanobiology Institute, Yale University, New Haven, United States.
Nanobiology Institute, Yale University, West Haven, United States.
Elife. 2021 May 13;10:e62084. doi: 10.7554/eLife.62084.
During clathrin-mediated endocytosis (CME) in eukaryotes, actin assembly is required to overcome large membrane tension and turgor pressure. However, the molecular mechanisms by which the actin machinery adapts to varying membrane tension remain unknown. In addition, how cells reduce their membrane tension when they are challenged by hypotonic shocks remains unclear. We used quantitative microscopy to demonstrate that cells rapidly reduce their membrane tension using three parallel mechanisms. In addition to using their cell wall for mechanical protection, yeast cells disassemble eisosomes to buffer moderate changes in membrane tension on a minute time scale. Meanwhile, a temporary reduction in the rate of endocytosis for 2-6 min and an increase in the rate of exocytosis for at least 5 min allow cells to add large pools of membrane to the plasma membrane. We built on these results to submit the cells to abrupt increases in membrane tension and determine that the endocytic actin machinery of fission yeast cells rapidly adapts to perform CME. Our study sheds light on the tight connection between membrane tension regulation, endocytosis, and exocytosis.
在真核生物的网格蛋白介导的胞吞作用(CME)中,肌动蛋白组装是克服大的膜张力和膨压差所必需的。然而,肌动蛋白机制如何适应变化的膜张力的分子机制尚不清楚。此外,当细胞受到低渗冲击时,它们如何降低膜张力也不清楚。我们使用定量显微镜技术证明,细胞通过三种平行的机制来快速降低膜张力。除了使用细胞壁进行机械保护外,酵母细胞还会解体埃斯莫斯体(eisosomes),以在分钟级的时间尺度上缓冲膜张力的适度变化。同时,短暂地将胞吞作用的速率降低 2-6 分钟,并将胞吐作用的速率至少增加 5 分钟,使细胞能够将大量的膜添加到质膜中。在此基础上,我们进一步让细胞突然增加膜张力,并确定裂殖酵母细胞的内吞作用肌动蛋白机制能够迅速适应 CME。我们的研究揭示了膜张力调节、内吞作用和胞吐作用之间的紧密联系。