Pennings S, Meersseman G, Bradbury E M
Department of Biological Chemistry, School of Medicine, University of California, Davis 95616.
J Mol Biol. 1991 Jul 5;220(1):101-10. doi: 10.1016/0022-2836(91)90384-i.
We report on a dynamic aspect of nucleosome positioning, in the absence of transcription-related events, on sea urchin 5 S rDNA. On tandem repeats of nucleosome length DNA of this strongly positioning sequence, histone octamers assemble in one dominant position surrounded by minor positions, ten base-pairs apart and therefore with identical rotational setting of the DNA coil. The existence of this cluster of positions, determined using micrococcal nuclease is confirmed by the results from DNase I footprinting and restriction enzyme analysis. The results from these techniques and from two-dimensional nucleoprotein polyacrylamide gel analysis indicate that the cluster of octamer positions is in dynamic equilibrium, in low ionic conditions, suggesting that the minor positions reflect fluctuations around the major nucleosome site. Histone octamer mobility appears to be temperature dependent and is reversibly inhibited by Mg2+.
我们报道了在没有转录相关事件的情况下,海胆5S核糖体DNA上核小体定位的一个动态方面。在这个强定位序列的核小体长度DNA的串联重复序列上,组蛋白八聚体组装在一个主要位置,周围是次要位置,相隔十个碱基对,因此DNA螺旋具有相同的旋转设置。使用微球菌核酸酶确定的这一位置簇的存在,通过DNA酶I足迹分析和限制性内切酶分析的结果得到了证实。这些技术以及二维核蛋白聚丙烯酰胺凝胶分析的结果表明,在低离子条件下,八聚体位置簇处于动态平衡,这表明次要位置反映了围绕主要核小体位点的波动。组蛋白八聚体的迁移率似乎与温度有关,并受到Mg2+的可逆抑制。