Davey C, Pennings S, Meersseman G, Wess T J, Allan J
Department of Biochemistry, University of Edinburgh, Scotland.
Proc Natl Acad Sci U S A. 1995 Nov 21;92(24):11210-4. doi: 10.1073/pnas.92.24.11210.
Positioned nucleosomes contribute to both the structure and the function of the chromatin fiber and can play a decisive role in controlling gene expression. We have mapped, at high resolution, the translational positions adopted by limiting amounts of core histone octamers reconstituted onto 4.4 kb of DNA comprising the entire chicken adult beta-globin gene, its enhancer, and flanking sequences. The octamer displays extensive variation in its affinity for different positioning sites, the range exhibited being about 2 orders of magnitude greater than that of the initial binding of the octamer. Strong positioning sites are located 5' and 3' of the globin gene and in the second intron but are absent from the coding regions. These sites exhibit a periodicity (approximately 200 bp) similar to the average spacing of nucleosomes on the inactive beta-globin gene in vivo, which could indicate their involvement in packaging the gene into higher-order chromatin structure. Overlapping, alternative octamer positioning sites commonly exhibit spacings of 20 and 40 bp, but not of 10 bp. These short-range periodicities could reflect features of the core particle structure contributing to the pronounced sequence-dependent manner in which the core histone octamer interacts with DNA.
定位核小体对染色质纤维的结构和功能均有贡献,并且在控制基因表达中可能起决定性作用。我们已在高分辨率下绘制了将有限数量的核心组蛋白八聚体重组到包含整个鸡成年β-珠蛋白基因、其增强子和侧翼序列的4.4 kb DNA上所采用的平移位置。八聚体对不同定位位点的亲和力表现出广泛差异,其表现出的范围比八聚体的初始结合范围大约大2个数量级。强定位位点位于珠蛋白基因的5'和3'端以及第二个内含子中,但编码区域没有。这些位点表现出与体内无活性β-珠蛋白基因上核小体的平均间距相似的周期性(约200 bp),这可能表明它们参与将基因包装成高阶染色质结构。重叠的、替代性的八聚体定位位点通常表现出20和40 bp的间距,但没有10 bp的间距。这些短程周期性可能反映了核心颗粒结构的特征,这些特征有助于核心组蛋白八聚体与DNA相互作用的明显序列依赖性方式。