An W, Zlatanova J, Leuba S H, van Holde K
Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331-7305, USA.
Biochimie. 1999 Jul;81(7):727-32. doi: 10.1016/s0300-9084(99)80130-8.
Using nucleosomes reconstituted on a defined sequence of DNA, we have investigated the question as to whether the N-terminal tails of core histones play a role in determining the site of binding of a linker histone. Reconstitutes used histone cores of three types: intact, lacking the N-terminal H3 tails, or lacking all tails. In each case the same, single defined position for the histone core was observed, using high-resolution mapping. The affinity for binding of linker histone H1(o) was highest for the intact cores, lowest for the tailless cores. However, the location of the linker histone, as judged by micrococcal nuclease protection, was exactly the same in each case, an asymmetric site of about 17 bp to one side of the core particle DNA.
利用在特定DNA序列上重构的核小体,我们研究了核心组蛋白的N端尾巴在决定连接组蛋白结合位点方面是否发挥作用这一问题。重构实验使用了三种类型的组蛋白核心:完整的、缺少N端H3尾巴的或缺少所有尾巴的。在每种情况下,通过高分辨率图谱分析观察到组蛋白核心的位置相同且单一。连接组蛋白H1(o)与完整核心的结合亲和力最高,与无尾核心的结合亲和力最低。然而,通过微球菌核酸酶保护判断,连接组蛋白在每种情况下的位置完全相同,位于核心颗粒DNA一侧约17 bp的不对称位点处。