Bionanoscience Department, Kavli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, 2628, CJ, The Netherlands.
Small. 2015 Feb 25;11(8):976-84. doi: 10.1002/smll.201401318. Epub 2014 Oct 21.
The accessibility of DNA is determined by the number, position, and stability of nucleosomes, complexes consisting of a core of 8 histone proteins with DNA wrapped around it. Since the structure and dynamics of nucleosomes affects essential cellular processes, they are the subject of many current studies. Here, high-speed atomic force microscopy is used to visualize dynamic processes in nucleosomes and tetrasomes (subnucleosomal structures that contain 4 rather than 8 histones in the protein core). Nucleosomes can spontaneously disassemble in a process (at a 1 second timescale). For tetrasomes, multiple dynamic phenomena are observed. For example, during disassembly the formation of a DNA loop (∼25 nm in length) is seen, which remains stable for several minutes. For intact tetrasomes, dynamics in the form of sliding and reversible hopping between stable positions along the DNA are observed. The data emphasize that tetrasomes are not merely static objects but highly dynamic. Since tetrasomes (in contrast to nucleosomes) can stay on the DNA during transcription, the observed tetrasome dynamics is relevant for an understanding of the nucleosomal dynamics during transcription. These results illustrate the diversity of nucleosome dynamics and demonstrate the ability of high speed AFM to characterize protein-DNA interactions.
DNA 的可及性由核小体的数量、位置和稳定性决定,核小体是由 8 个核心组蛋白与包裹在其周围的 DNA 组成的复合物。由于核小体的结构和动力学影响着基本的细胞过程,因此它们是许多当前研究的主题。在这里,高速原子力显微镜被用于可视化核小体和四联体(包含核心蛋白中 4 个而不是 8 个组蛋白的亚核小体结构)中的动态过程。核小体可以在一个过程中(在 1 秒的时间尺度上)自发地解体。对于四联体,观察到多种动态现象。例如,在解体过程中,形成了一个 DNA 环(长度约为 25nm),它可以稳定几分钟。对于完整的四联体,观察到沿着 DNA 滑动和在稳定位置之间可逆跳跃的动力学。这些数据强调了四联体不仅仅是静态的物体,而是高度动态的。由于四联体(与核小体相比)在转录过程中可以留在 DNA 上,因此观察到的四联体动力学对于理解转录过程中核小体的动力学是相关的。这些结果说明了核小体动力学的多样性,并展示了高速原子力显微镜在表征蛋白质-DNA 相互作用方面的能力。