...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Cell cycle >Biotin-tag affinity purification of a centromeric nucleosome assembly complex.
【24h】

Biotin-tag affinity purification of a centromeric nucleosome assembly complex.

机译：着丝粒核小体装配复合物的生物素标签亲和纯化。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Centromeres are chromosomal sites of microtubule binding that ensure correct mitotic segregation of chromosomes to daughter cells. This process is mediated by a special centromere-specific histone H3 variant (CenH3), which packages centromeric chromatin and epigenetically maintains the centromere at a distinct chromosomal location. However, CenH3 is present at low abundance relative to canonical histones, presenting a challenge for the isolation and characterization of the chaperone machinery that assembles CenH3 into nucleosomes at centromeres. To address this challenge, we used controlled overexpression of Drosophila CenH3 (CID) and an efficient biochemical purification strategy offered by in vivo biotinylation of CID to successfully purify and characterize the soluble CID nucleosome assembly complex. It consists of a single chaperone protein, RbAp48, complexed with CID and histone H4. RbAp48 is also found in protein complexes that assemble canonical histone H3 and replacement histone H3.3. Here, we highlight the benefits of our improved biotin-mediated purification method, and address the question of how the simple CID/H4-RbAp48 chaperone complex can mediate nucleosome assembly specifically at centromeres.
机译：着丝粒是微管结合的染色体位点，可确保染色体与子细胞的正确有丝分裂分离。此过程由特殊的着丝粒特异性组蛋白H3变体（CenH3）介导，该变体包装着丝粒染色质并在表观遗传上将着丝粒保持在不同的染色体位置。然而，相对于规范组蛋白，CenH3的丰度低，这对分离和表征将着丝粒CenH3组装成核小体的分子伴侣机制提出了挑战。为了解决这一挑战，我们使用了果蝇CenH3（CID）的受控过表达和CID体内生物素化提供的有效生化纯化策略，以成功地纯化和表征可溶性CID核小体装配复合物。它由单一的伴侣蛋白RbAp48与CID和组蛋白H4复合而成。 RbAp48还存在于蛋白质复合物中，该复合物可组装典型的组蛋白H3和替代组蛋白H3.3。在这里，我们着重介绍改进的生物素介导的纯化方法的优势，并解决简单的CID / H4-RbAp48分子伴侣复合物如何介导着丝粒特别是着丝粒的问题。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号