Hayes J J, Tullius T D, Wolffe A P
Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1990 Oct;87(19):7405-9. doi: 10.1073/pnas.87.19.7405.
We describe the application of the hydroxyl radical footprinting technique to examine the histone-DNA interactions of a nucleosome that includes part of the 5S ribosomal RNA gene of Xenopus borealis. We establish that two distinct regions of DNA with different helical periodicities exist within the nucleosome and demonstrate a change in the helical periodicity of this DNA upon nucleosome formation. In particular, we find that on average the helical periodicity of DNA in this nucleosome is 10.18 +/- 0.05 base pairs per turn. The same DNA, when bound to a calcium phosphate surface, has a periodicity of 10.49 +/- 0.05 base pairs per turn, similar to that of random sequence DNA. Modulations in minor groove width within the naked DNA detected by the hydroxyl radical are maintained and exaggerated in nucleosomal DNA. These features correlate with regions in the DNA previously suggested to be important for nucleosome positioning.
我们描述了羟基自由基足迹技术在研究包含北极爪蟾5S核糖体RNA基因一部分的核小体的组蛋白-DNA相互作用中的应用。我们确定在核小体内存在两个具有不同螺旋周期性的不同DNA区域,并证明了该DNA在核小体形成时螺旋周期性的变化。特别地,我们发现该核小体中DNA的平均螺旋周期性为每圈10.18±0.05个碱基对。相同的DNA与磷酸钙表面结合时,每圈的周期性为10.49±0.05个碱基对,与随机序列DNA相似。羟基自由基检测到的裸露DNA中小沟宽度的调制在核小体DNA中得以维持并被放大。这些特征与先前认为对核小体定位很重要的DNA区域相关。