A comparison of in vitro nucleosome positioning mapped with chicken, frog and a variety of yeast core histones.
A comparison of in vitro nucleosome positioning mapped with chicken, frog and a variety of yeast core histones.
复制标题
DOI：
10.1016/j.jmb.2013.07.019
复制
发表时间：
2013-11-15
影响因子：
5.6
通讯作者：
Singh V
中科院分区：
文献类型：
--
作者：
Allan J;Fraser RM;Owen-Hughes T;Docherty K;Singh V
利用高通量测序，我们在四个含有来自绵羊、果蝇、人和酵母基因组的DNA片段的质粒DNA上定位了体外序列定向的核小体定位。以鸡、蛙和酵母核心组蛋白为原料，通过重组法制备了色素。我们还组装了酵母染色质，其中组蛋白H3被着丝粒特异的组蛋白变体Cse4取代。重组蛙和土鸡组蛋白的位置非常相似。相比之下，含有规范的酵母八聚体，特别是Cse4八聚体的核小体在不同的位置组装，这一特性在特定的基因组DNA片段上更加明显。文中还讨论了核小体定位变化的可能因素和行为的含义。用四种组蛋白八聚体在含有绵羊、果蝇、人和酵母基因组DNA片段的质粒上形成染色质。每一次重组都要测量核小体的位置。虽然相似，但用酵母组蛋白八聚体获得的结合图谱，包括一个含有Cse4的结合图谱，与用鸡或青蛙组蛋白获得的结合图谱不同。核小体定位的差异在富含AT的DNA底物上表现得最明显。
Using high-throughput sequencing, we have mapped sequence-directed nucleosome positioning in vitro on four plasmid DNAs containing DNA fragments derived from the genomes of sheep, drosophila, human and yeast. Chromatins were prepared by reconstitution using chicken, frog and yeast core histones. We also assembled yeast chromatin in which histone H3 was replaced by the centromere-specific histone variant, Cse4. The positions occupied by recombinant frog and native chicken histones were found to be very similar. In contrast, nucleosomes containing the canonical yeast octamer or, in particular, the Cse4 octamer were assembled at distinct populations of locations, a property that was more apparent on particular genomic DNA fragments. The factors that may contribute to this variation in nucleosome positioning and the implications of the behavior are discussed. Chromatins were formed on plasmids containing fragments of sheep, drosophila, human and yeast genomic DNAs with four types of histone octamer. Nucleosome positioning was measured for each reconstitute. Although similar, the binding profiles obtained with the yeast histone octamer, including one containing Cse4, were distinct from those obtained with chick or frog histones. The difference in nucleosome positioning was best seen on AT-rich DNA substrates.