Pasolli Milena, Meiring Joyce C M, Conboy James P, Koenderink Gijsje H, Akhmanova Anna
Cell Biology, Neurobiology and Biophysics, Department of Biology, Faculty of Science, Utrecht University, Utrecht, The Netherlands.
Department of Bionanoscience, Kavli Institute of Nanoscience Delft, Delft University of Technology, HZ Delft, The Netherlands.
J Cell Biol. 2025 Sep 1;224(9). doi: 10.1083/jcb.202504004. Epub 2025 Jul 8.
Intermediate filaments (IFs) are a key component of the cytoskeleton, essential for regulating cell mechanics, maintaining nuclear integrity, organelle positioning, and modulating cell signaling. Current insights into IF function primarily come from studies using long-term perturbations, such as protein depletion or mutation. Here, we present tools that allow rapid manipulation of vimentin IFs in the whole cytoplasm or within specific subcellular regions by inducibly coupling them to microtubule motors, either pharmacologically or using light. Rapid perinuclear clustering of vimentin had no major immediate effects on the actin or microtubule organization, cell spreading, or focal adhesion number, but it reduced cell stiffness. Mitochondria and endoplasmic reticulum (ER) sheets were reorganized due to vimentin clustering, whereas lysosomes were only briefly displaced and rapidly regained their normal distribution. Keratin moved along with vimentin in some cell lines but remained intact in others. Our tools help to study the immediate and local effects of vimentin perturbation and identify direct links of vimentin to other cellular structures.
中间丝（IFs）是细胞骨架的关键组成部分，对于调节细胞力学、维持核完整性、细胞器定位以及调节细胞信号传导至关重要。目前对中间丝功能的认识主要来自使用长期扰动的研究，例如蛋白质缺失或突变。在此，我们展示了一些工具，这些工具能够通过药理学方法或利用光将波形蛋白中间丝与微管马达诱导偶联，从而在整个细胞质或特定亚细胞区域快速操纵波形蛋白中间丝。波形蛋白在核周的快速聚集对肌动蛋白或微管组织、细胞铺展或粘着斑数量没有主要的即时影响，但它降低了细胞硬度。线粒体和内质网（ER）片层因波形蛋白聚集而重新组织，而溶酶体仅被短暂移位并迅速恢复其正常分布。角蛋白在一些细胞系中与波形蛋白一起移动，但在其他细胞系中保持完整。我们的工具有助于研究波形蛋白扰动的即时和局部影响，并确定波形蛋白与其他细胞结构的直接联系。
