Physical Plasma Membrane Perturbation Using Subcellular Optogenetics Drives Integrin-Activated Cell Migration
Physical Plasma Membrane Perturbation Using Subcellular Optogenetics Drives Integrin-Activated Cell Migration
复制标题
DOI：
10.1021/acssynbio.8b00356
复制
发表时间：
2019-03-01
影响因子：
4.7
通讯作者：
Gautam, N.
中科院分区：
文献类型：
--
作者：
Meshik, Xenia;O'Neill, Patrick R.;Gautam, N.
细胞经历质膜的物理变形，这可以调节细胞行为，如迁移。了解物理线索如何影响动态细胞反应的分子基础，需要新的方法，可以物理扰动质膜快速，可逆，亚细胞控制。在这里，我们提出了一种基于光诱导二聚化的光遗传学方法，该方法通过将高浓度的胞质蛋白募集到靶位点来改变质膜特性。令人惊讶的是，细胞中蛋白质的这种极化积累诱导了相反方向的定向变形虫迁移。与体外将高浓度蛋白质限制在膜上的已知效应一致，质膜中存在局部曲率和张力降低。对机械力敏感的整合素活性在该区域被激活。整合素与光遗传学的局部机械激活允许分子和细胞反应的同时成像，有助于揭示正反馈回路，包括SFK和ERK依赖性RhoA激活，肌动球蛋白收缩性，向后膜流动和膜张力降低，这是细胞迁移模式的基础。
Cells experience physical deformations to the plasma membrane that can modulate cell behaviors like migration. Understanding the molecular basis for how physical cues affect dynamic cellular responses requires new approaches that can physically perturb the plasma membrane with rapid, reversible, subcellular control. Here we present an optogenetic approach based on light-inducible dimerization that alters plasma membrane properties by recruiting cytosolic proteins at high concentrations to a target site. Surprisingly, this polarized accumulation of proteins in a cell induces directional amoeboid migration in the opposite direction. Consistent with known effects of constraining high concentrations of proteins to a membrane in vitro, there is localized curvature and tension decrease in the plasma membrane. Integrin activity, sensitive to mechanical forces, is activated in this region. Localized mechanical activation of integrin with optogenetics allowed simultaneous imaging of the molecular and cellular response, helping uncover a positive feedback loop comprising SFK- and ERK-dependent RhoA activation, actomyosin contractility, rearward membrane flow, and membrane tension decrease underlying this mode of cell migration.