...
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Stepwise loading of yeast clamp revealed by ensemble and single-molecule studies
【24h】

Stepwise loading of yeast clamp revealed by ensemble and single-molecule studies

机译:整体和单分子研究揭示了酵母夹的逐步加载

获取原文
获取原文并翻译 | 示例
           

摘要

In ensemble and single-molecule experiments using the yeast proliferating cell nuclear antigen (PCNA, clamp) and replication factor C (RFC, clamp loader), we have examined the assembly of the RFC ? PCNA · DNA complex and its progression to holoenzyme upon addition of polymerase 5 (pol5). We obtained data that indicate (Ⅰ) PCNA loading on DNA proceeds through multiple conforma-tional intermediates and is successful after several failed attempts; (Ⅱ) RFC does not act catalytically on a primed 45-mer templated fork; (Ⅲ) the RFC ? PCNA · DNA complex formed in the presence of ATP is derived from at least two kinetically distinguishable species; (/V) these species disassemble through either unloading of RFC ? PCNA from DNA or dissociation of PCNA into its component subunits; and (v) in the presence of polδ only one species converts to the RFC · PCNA · DNA · polδ holoenzyme. These findings redefine and deepen our understanding of the clamp-loading process and reveal that it is surprisingly one of trial and error to arrive at a heuristic solution.
机译:在使用酵母增殖细胞核抗原(PCNA,钳位)和复制因子C(RFC,钳位加载器)的整体和单分子实验中,我们检查了RFC? PCNA·DNA复合物及其添加聚合酶5(pol5)后向全酶的发展。我们获得的数据表明(Ⅰ)DNA上PCNA的装载是通过多种构象中间体进行的,并且经过几次失败的尝试才成功。 (Ⅱ)RFC对涂有底漆的45-mer模板叉没有催化作用; (三)RFC?在ATP存在下形成的PCNA·DNA络合物至少来自两个动力学上可区分的物种。 (/ V)这些种类通过RFC的卸载而分解。 DNA中的PCNA或PCNA分解成其组成亚基; (v)在存在polδ的情况下,只有一种会转化为RFC·PCNA·DNA·polδ全酶。这些发现重新定义并加深了我们对夹具加载过程的理解,并揭示出启发式解决方案出奇地是反复试验之一。

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