DNA AND PROTEIN DETERMINANTS OF NUCLEOSOME POSITIONING ON SEA-URCHIN 5S RIBOSOMAL-RNA GENE-SEQUENCES INVITRO
DNA AND PROTEIN DETERMINANTS OF NUCLEOSOME POSITIONING ON SEA-URCHIN 5S RIBOSOMAL-RNA GENE-SEQUENCES INVITRO
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DOI：
10.1073/pnas.87.15.5724
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发表时间：
1990-08-01
影响因子：
11.1
通讯作者：
VANHOLDE, KE
中科院分区：
文献类型：
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作者：
DONG, F;HANSEN, JC;VANHOLDE, KE
核小体定位的DNA和蛋白质的决定因素已被检查后，在体外重组的天然或修饰的组蛋白八聚体的串联重复序列的207-和172-碱基对的DNA序列包含的Lytechinus variegatus 5S rRNA基因和单体序列来自这些消化与各种限制性内切酶。在所有情况下，观察到一个主要的核小体位置以及一些次要位置，这表明多个位置的产生是5S rRNA基因序列的固有特性。有趣的是，观察到的所有位置都相差10个碱基对的倍数。在不同的重组条件下获得的数据表明，所观察到的核小体在这些DNA模板上的分布是平衡分布。本研究还研究了组蛋白八聚体的定位，其中组蛋白“尾巴”已被胰蛋白酶消化去除。结果表明，组蛋白尾部不是核小体定位的决定因素。虽然我们的研究结果表明，5S rRNA的机械特性是决定核小体定位的基本因素，但它们不足以将所有核小体引导到单个位置。
DNA and protein determinants of nucleosome positioning have been examined after in vitro reconstitutions of native or modified histone octamers onto tandem repeats of 207- and 172-base-pair DNA sequences containing the Lytechinus variegatus 5S rRNA gene and onto monomeric sequences derived from these by digestion with various restriction endonucleases. In all cases, a major nucleosome position as well as a number of minor positions have been observed, which indicates that the generation of multiple positions is an inherent property of the 5S rRNA gene sequence. Interestingly, all positions observed differ by multiples of 10 base pairs. Data obtained under different reconstitution conditions demonstrate that the observed distributions of nucleosomes on these DNA templates are equilibrium distributions. This study has also examined the positioning of histone octamers from which histone "tails" had been removed by tryptic digestion. Results indicate that the histone tails are not determinants of nucleosome positioning. Although our results suggest that the mechanical properties of the 5S rRNA are the fundamental factors determining nucleosome positioning, they are insufficient to direct all nucleosomes into a single location.