University of California, San Francisco, San Francisco, CA 94158, USA.
Mol Biol Cell. 2013 Apr;24(7):982-94. doi: 10.1091/mbc.E12-09-0685. Epub 2013 Feb 27.
In addition to its long-studied presence in the cytoplasm, actin is also found in the nuclei of eukaryotic cells. The function and form (monomer, filament, or noncanonical oligomer) of nuclear actin are hotly debated, and its localization and dynamics are largely unknown. To determine the distribution of nuclear actin in live somatic cells and evaluate its potential functions, we constructed and validated fluorescent nuclear actin probes. Monomeric actin probes concentrate in nuclear speckles, suggesting an interaction of monomers with RNA-processing factors. Filamentous actin probes recognize discrete structures with submicron lengths that are excluded from chromatin-rich regions. In time-lapse movies, these actin filament structures exhibit one of two types of mobility: 1) diffusive, with an average diffusion coefficient of 0.06-0.08 μm(2)/s, or (2) subdiffusive, with a mobility coefficient of 0.015 μm(2)/s. Individual filament trajectories exhibit features of particles moving within a viscoelastic mesh. The small size of nuclear actin filaments is inconsistent with a role in micron-scale intranuclear transport, and their localization suggests that they do not participate directly in chromatin-based processes. Our results instead suggest that actin filaments form part of a large, viscoelastic structure in the nucleoplasm and may act as scaffolds that help organize nuclear contents.
除了在细胞质中被长期研究外,肌动蛋白也存在于真核细胞的核内。核内肌动蛋白的功能和形式(单体、纤维或非典型寡聚物)存在激烈争议,其定位和动态变化在很大程度上仍是未知的。为了确定核内肌动蛋白在活体体细胞中的分布并评估其潜在功能,我们构建并验证了荧光核肌动蛋白探针。单体肌动蛋白探针集中在核斑点中,表明单体与 RNA 处理因子相互作用。丝状肌动蛋白探针识别具有亚微米长度的离散结构,这些结构不与富含染色质的区域重叠。在延时电影中,这些肌动蛋白纤维结构表现出两种运动类型之一:1)扩散性,平均扩散系数为 0.06-0.08 μm²/s,或 2)亚扩散性,迁移系数为 0.015 μm²/s。单个纤维轨迹表现出在粘弹性网格中移动的粒子的特征。核内肌动蛋白纤维的小尺寸与微米尺度的核内运输作用不一致,其定位表明它们不直接参与基于染色质的过程。我们的结果表明,肌动蛋白纤维形成核质中一个大的粘弹性结构的一部分,并且可能作为帮助组织核内容物的支架。