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Expansion of Chromosome Territories with Chromatin Decompaction in BAF53-depleted Interphase Cells

机译:贫乏BAF53间期细胞中的染色质失配与染色体领土的扩大。

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摘要

Chromosomes are compartmentalized into discrete chromosome territories during interphase in mammalian cells. A chromosome territory is generated by the tendency of chromatin to occupy the smallest shell volume, which is determined by the polymeric properties and interactions of the internal meshwork of the chromatin fiber. Here, we show that BAF53 knockdown by small interfering RNA interference led to the expansion of chromosome territories. This was accompanied by a reduction in chromatin compaction, an increase in the micrococcal nuclease sensitivity of the chromatin, and an alteration in H3-K9 and H3-K79 dimethylation. Interestingly, the BAF53 knockdown cells suffer a cell cycle defect. Despite the significant irregularity and decompaction of the polynucleosomes isolated from the BAF53 knockdown cells, the chromatin loading of H1 and core histones remained unaltered, as did the nucleosome spacing. The histone hyperacetylation and down-regulation of BRG-1, mBrm, and Tip49, the catalytic components of the SWI/SNF complex and the TIP60 complex, respectively, did not expand chromosome territories. These results indicate that BAF53 contributes to the polymeric properties and/or the internal meshwork interactions of the chromatin fiber probably via a novel mechanism.
机译:在哺乳动物细胞的相间期,染色体被划分为离散的染色体区域。染色体区域是由染色质占据最小外壳体积的趋势生成的,该趋势由染色质纤维的聚合特性和内部网状结构的相互作用决定。在这里,我们表明小干扰RNA干扰BAF53击倒导致染色体领土的扩大。这伴随着染色质紧实度的降低,染色质的微球菌核酸酶敏感性的增加以及H3-K9和H3-K79二甲基化的改变。有趣的是,BAF53敲低细胞遭受细胞周期缺陷。尽管从BAF53敲低细胞中分离出的多核小体存在明显的不规则性和分解性,但H1和核心组蛋白的染色质负载仍保持不变,核小体间距也保持不变。 BRWI-1,mBrm和Tip49的组蛋白高度乙酰化和下调(分别是SWI / SNF复合体和TIP60复合体的催化成分)没有扩展染色体区域。这些结果表明BAF53可能通过一种新颖的机制对染色质纤维的聚合物性质和/或内部网孔相互作用做出了贡献。

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