Biological Macromolecules Laboratory, Structural Biology Center, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Cell Rep. 2012 Dec 27;2(6):1645-56. doi: 10.1016/j.celrep.2012.11.008. Epub 2012 Dec 13.
Genome information, which is three-dimensionally organized within cells as chromatin, is searched and read by various proteins for diverse cell functions. Although how the protein factors find their targets remains unclear, the dynamic and flexible nature of chromatin is likely crucial. Using a combined approach of fluorescence correlation spectroscopy, single-nucleosome imaging, and Monte Carlo computer simulations, we demonstrate local chromatin dynamics in living mammalian cells. We show that similar to interphase chromatin, dense mitotic chromosomes also have considerable chromatin accessibility. For both interphase and mitotic chromatin, we observed local fluctuation of individual nucleosomes (~50 nm movement/30 ms), which is caused by confined Brownian motion. Inhibition of these local dynamics by crosslinking impaired accessibility in the dense chromatin regions. Our findings show that local nucleosome dynamics drive chromatin accessibility. We propose that this local nucleosome fluctuation is the basis for scanning genome information.
基因组信息作为染色质在细胞内三维组织,被各种蛋白搜索和读取,以实现多种细胞功能。虽然蛋白因子如何找到其靶标仍不清楚,但染色质的动态和灵活性质可能是至关重要的。我们采用荧光相关光谱法、单核小体成像和蒙特卡罗计算机模拟的联合方法,在活的哺乳动物细胞中展示了局部染色质动力学。我们表明,类似于间期染色质,密集的有丝分裂染色体也具有相当大的染色质可及性。对于间期和有丝分裂染色质,我们观察到单个核小体的局部波动(~50nm 运动/30ms),这是由受限的布朗运动引起的。交联抑制这些局部动力学会损害致密染色质区域的可及性。我们的发现表明,局部核小体动力学驱动染色质可及性。我们提出,这种局部核小体波动是扫描基因组信息的基础。