Center for Soft Matter Research, Department of Physics, New York University, New York, NY 10003.
Center for Soft Matter Research, Department of Physics, New York University, New York, NY 10003
Proc Natl Acad Sci U S A. 2017 Sep 26;114(39):10338-10343. doi: 10.1073/pnas.1702226114. Epub 2017 Sep 12.
The nuclear envelope (NE) presents a physical boundary between the cytoplasm and the nucleoplasm, sandwiched in between two highly active systems inside the cell: cytoskeleton and chromatin. NE defines the shape and size of the cell nucleus, which increases during the cell cycle, accommodating for chromosome decondensation followed by genome duplication. In this work, we study nuclear shape fluctuations at short time scales of seconds in human cells. Using spinning disk confocal microscopy, we observe fast fluctuations of the NE, visualized by fluorescently labeled lamin A, and of the chromatin globule surface (CGS) underneath the NE, visualized by fluorescently labeled histone H2B. Our findings reveal that fluctuation amplitudes of both CGS and NE monotonously decrease during the cell cycle, serving as a reliable cell cycle stage indicator. Remarkably, we find that, while CGS and NE typically fluctuate in phase, they do exhibit localized regions of out-of-phase motion, which lead to separation of NE and CGS. To explore the mechanism behind these shape fluctuations, we use biochemical perturbations. We find the shape fluctuations of CGS and NE to be both thermally and actively driven, the latter caused by forces from chromatin and cytoskeleton. Such undulations might affect gene regulation as well as contribute to the anomalously high rates of nuclear transport by, e.g., stirring of molecules next to NE, or increasing flux of molecules through the nuclear pores.
核膜(NE)在细胞质和核质之间提供了一个物理边界,夹在细胞内两个高度活跃的系统之间:细胞骨架和染色质。NE 定义了细胞核的形状和大小,在细胞周期中会增加,以容纳染色体解凝聚和基因组复制。在这项工作中,我们研究了人类细胞中秒级短时间尺度的核形状波动。使用旋转盘共聚焦显微镜,我们观察到由荧光标记的核纤层 A 可视化的 NE 和在其下的染色质球表面(CGS)的快速波动,由荧光标记的组蛋白 H2B 可视化。我们的发现表明,CGS 和 NE 的波动幅度在细胞周期中单调减小,可作为可靠的细胞周期阶段指标。值得注意的是,我们发现,虽然 CGS 和 NE 通常相位波动,但它们确实表现出局部的非相位运动区域,导致 NE 和 CGS 分离。为了探索这些形状波动的机制,我们使用生化扰动。我们发现 CGS 和 NE 的形状波动既受热驱动又受主动驱动,后者是由染色质和细胞骨架的力引起的。这种波动可能会影响基因调控,并通过例如在 NE 附近搅拌分子或增加通过核孔的分子通量,为异常高的核转运速率做出贡献。