...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >The WYL Domain of the PIF1 Helicase from the Thermophilic Bacterium Thermotoga elfii is an Accessory Single-Stranded DNA Binding Module
【24h】

The WYL Domain of the PIF1 Helicase from the Thermophilic Bacterium Thermotoga elfii is an Accessory Single-Stranded DNA Binding Module

机译：来自嗜热细菌的PIF1螺旋酶的Wyl结构域热敏胶酶热湿度是辅助单链DNA装订模块

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

PIF1 family helicases are conserved from bacteria to man. With the exception of the well-studied yeast PIF1 helicases (e.g., ScPif1 and ScRrm3), however, very little is known about how these enzymes help maintain genome stability. Indeed, we lack a basic understanding of the protein domains found N- and C-terminal to the characteristic central PIF1 helicase domain in these proteins. Here, using chimeric constructs, we show that the ScPif1 and ScRrm3 helicase domains are interchangeable and that the N-terminus of ScRrm3 is important for its function in vivo . This suggests that PIF1 family helicases evolved functional modules fused to a generic motor domain. To investigate this hypothesis, we characterized the biochemical activities of the PIF1 helicase from the thermophilic bacterium Thermotoga elfii (TePif1), which contains a C-terminal WYL domain of unknown function. Like helicases from other thermophiles, recombinant TePif1 was easily prepared, thermostable in vitro , and displayed activities similar to its eukaryotic homologues. We also found that the WYL domain was necessary for high-affinity single-stranded DNA (ssDNA) binding and affected both ATPase and helicase activities. Deleting the WYL domain from TePif1 or mutating conserved residues in the predicted ssDNA binding site uncoupled ATPase activity and DNA unwinding, leading to higher rates of ATP hydrolysis but less efficient DNA helicase activity. Our findings suggest that the domains of unknown function found in eukaryotic PIF1 helicases may also confer functional specificity and additional activities to these enzymes, which should be investigated in future work.
机译：PIF1家庭螺旋酶从细菌保存到男人身上。然而，除了研究良好的酵母PIF1螺旋酶（例如，SCPIF1和SCRRM3）之外，关于这些酶如何有助于维持基因组稳定性的情况很少。实际上，我们缺乏对这些蛋白质中的特征中央PIF1螺旋酶结构域发现N-和C末端的基本理解。这里，使用嵌合构建体，我们表明SCPIF1和SCRRM3旋光酶结构域是可互换的，并且SCRRM3的N-末端对于其在体内的功能很重要。这表明PIF1家族螺旋酶进化到融合到通用电机域的功能模块。为了研究该假设，我们将PIF1螺旋酶的生物化学活性从嗜热嗜热菌热量（TEPIF1）的表征为含有未知功能的C末端Wyl结构域。与来自其他嗜热料的螺旋酶一样，重组TEPIF1易于制备，体外热稳定，并显示出类似于其真核同源物的活动。我们还发现，Wyl结构域是高亲和力单链DNA（SSDNA）结合并影响ATP酶和螺旋酶活性的必要条件。从TEPIF1中删除Wyl结构域或在预测的SSDNA结合位点中突变保守的残留物，未替换的ATP酶活性和DNA展开，导致较高的ATP水解速率，但较低的DNA螺旋酶活性。我们的研究结果表明，真核PIF1螺旋酶中发现的未知功能的结构域也可能赋予这些酶的功能特异性和额外的活动，这应该在将来的工作中调查。

著录项

  • 来源
    《Biochemistry》 |2018年第7期|共11页
  • 作者

    Nicholas M. Andis; Christopher W. Sausen; Ashna Alladin; Matthew L. Bochman;

  • 作者单位

    Molecular and Cellular Biochemistry Department Indiana University Bloomington Indiana 47405 United States;

    Molecular and Cellular Biochemistry Department Indiana University Bloomington Indiana 47405 United States;

    Molecular and Cellular Biochemistry Department Indiana University Bloomington Indiana 47405 United States;

    Molecular and Cellular Biochemistry Department Indiana University Bloomington Indiana 47405 United States;

  • 收录信息
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物化学;
  • 关键词

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