Horowitz R A, Agard D A, Sedat J W, Woodcock C L
Department of Biology, University of Massachusetts, Amherst 01003.
J Cell Biol. 1994 Apr;125(1):1-10. doi: 10.1083/jcb.125.1.1.
The three dimensional (3D) structure of chromatin fibers in sections of nuclei has been determined using electron tomography. Low temperature embedding and nucleic acid-specific staining allowed individual nucleosomes to be clearly seen, and the tomographic data collection parameters provided a reconstruction resolution of 2.5 nm. Chromatin fibers have complex 3D trajectories, with smoothly bending regions interspersed with abrupt changes in direction, and U turns. Nucleosomes are located predominantly at the fiber periphery, and linker DNA tends to project toward the fiber interior. Within the fibers, a unifying structural motif is a two nucleosome-wide ribbon that is variably bent and twisted, and in which there is little face-to-face contact between nucleosomes. It is suggested that this asymmetric 3D zig-zag of nucleosomes and linker DNA represents a basic principle of chromatin folding that is determined by the properties of the nucleosome-linker unit. This concept of chromatin fiber architecture is contrasted with helical models in which specific nucleosome-nucleosome contacts play a major role in generating a symmetrical higher order structure. The transcriptional control implications of a more open and irregular chromatin structure are discussed.
利用电子断层扫描技术已确定了细胞核切片中染色质纤维的三维(3D)结构。低温包埋和核酸特异性染色使得单个核小体清晰可见,断层扫描数据采集参数提供了2.5纳米的重建分辨率。染色质纤维具有复杂的3D轨迹,有平滑弯曲的区域,其间穿插着方向的突然变化和U形转弯。核小体主要位于纤维周边,连接DNA倾向于向纤维内部突出。在纤维内部,一个统一的结构基序是一条由两个核小体宽的带,其弯曲和扭曲程度各不相同,并且核小体之间几乎没有面对面的接触。有人提出,核小体和连接DNA的这种不对称3D之字形结构代表了染色质折叠的一个基本原理,该原理由核小体 - 连接体单元的特性决定。这种染色质纤维结构的概念与螺旋模型形成对比,在螺旋模型中,特定的核小体 - 核小体接触在产生对称的高阶结构中起主要作用。文中还讨论了更开放和不规则的染色质结构对转录控制的影响。