...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>FEBS Open Bio >Proliferating cell nuclear antigen restores the enzymatic activity of a DNA ligase I deficient in DNA binding
【24h】

Proliferating cell nuclear antigen restores the enzymatic activity of a DNA ligase I deficient in DNA binding

机译：增殖的细胞核抗原恢复了DNA结合不足的DNA连接酶I的酶促活性

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Proliferating cell nuclear antigen (PCNA) coordinates multienzymatic reactions by interacting with a variety of protein partners. Family I DNA ligases are multidomain proteins involved in sealing of DNA nicks during Okazaki fragment maturation and DNA repair. The interaction of DNA ligases with the interdomain connector loop (IDCL) of PCNA through its PCNA‐interacting peptide (PIP box) is well studied but the role of the interacting surface between both proteins is not well characterized. In this work, we used a minimal DNA ligase I and two N‐terminal deletions to establish that DNA binding and nick‐sealing stimulation of DNA ligase I by PCNA are not solely dependent on the PIP box–IDCL interaction. We found that a truncated DNA ligase I with a deleted PIP box is stimulated by PCNA. Furthermore, the activity of a DNA ligase defective in DNA binding is rescued upon PCNA addition. As the rate constants for single‐turnover ligation for the full‐length and truncated DNA ligases are not affected by PCNA, our data suggest that PCNA stimulation is achieved by increasing the affinity for nicked DNA substrate and not by increasing catalytic efficiency. Surprisingly C‐terminal mutants of PCNA are not able to stimulate nick‐sealing activity of Entamoeba histolytica DNA ligase I. Our data support the notion that the C‐terminal region of PCNA may be involved in promoting an allosteric transition in E. histolytica DNA ligase I from a spread‐shaped to a ring‐shaped structure. This study suggests that the ring‐shaped PCNA is a binding platform able to stabilize coevolved protein–protein interactions, in this case an interaction with DNA ligase I.
机译：增殖细胞核抗原（PCNA）通过与多种蛋白质伴侣相互作用来协调多酶反应。家族I DNA连接酶是在Okazaki片段成熟和DNA修复过程中涉及DNA缺口密封的多域蛋白。 DNA连接酶通过其PCNA相互作用肽（PIP盒）与PCNA的域间连接环（IDCL）之间的相互作用已得到了很好的研究，但两种蛋白之间相互作用表面的作用尚不十分清楚。在这项工作中，我们使用了最小的DNA连接酶I和两个N末端缺失来确定PCNA的DNA结合和对DNA连接酶I的切口密封刺激并不仅仅依赖于PIP box-IDCL相互作用。我们发现PCNA刺激具有删除的PIP框的截短的DNA连接酶I。此外，添加PCNA后可以挽救DNA结合缺陷的DNA连接酶的活性。由于全长和截短的DNA连接酶的单周转连接速率常数不受PCNA的影响，我们的数据表明PCNA刺激是通过增加对带切口的DNA底物的亲和力而不是通过提高催化效率来实现的。出乎意料的是，PCNA的C末端突变体不能刺激组织解脂Entamoeba DNA连接酶I的切口封闭活性。我从散布型到环形结构。这项研究表明，环形PCNA是能够稳定共同进化的蛋白质间相互作用的结合平台，在这种情况下，这种相互作用是与DNA连接酶I相互作用的。

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