...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>The FEBS journal >Molecular modeling and functional characterization of the monomeric primase-polymerase domain from the Sulfolobus solfataricus plasmid pIT3.
【24h】

Molecular modeling and functional characterization of the monomeric primase-polymerase domain from the Sulfolobus solfataricus plasmid pIT3.

机译：来自Sulfolobus solfataricus质粒pIT3的单体引发酶-聚合酶结构域的分子建模和功能表征。

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

A tri-functional monomeric primase-polymerase domain encoded by the plasmid pIT3 from Sulfolobus solfataricus strain IT3 was identified using a structural-functional approach. The N-terminal domain of the pIT3 replication protein encompassing residues 31-245 (i.e. Rep245) was modeled onto the crystallographic structure of the bifunctional primase-polymerase domain of the archaeal plasmid pRN1 and refined by molecular dynamics in solution. The Rep245 protein was purified following overexpression in Escherichia coli and its nucleic acid synthesis activity was characterized. The biochemical properties of the polymerase activity such as pH, temperature optima and divalent cation metal dependence were described. Rep245 was capable of utilizing both ribonucleotides and deoxyribonucleotides for de novo primer synthesis and it synthesized DNA products up to several kb in length in a template-dependent manner. Interestingly, the Rep245 primase-polymerase domain harbors also a terminal nucleotidyl transferase activity, being able to elongate the 3'-end of synthetic oligonucleotides in a non-templated manner. Comparative sequence-structural analysis of the modeled Rep245 domain with other archaeal primase-polymerases revealed some distinctive features that could account for the multifaceted activities exhibited by this domain. To the best of our knowledge, Rep245 typifies the shortest functional domain from a crenarchaeal plasmid endowed with DNA and RNA synthesis and terminal transferase activity.
机译：使用结构功能方法鉴定了由Sulfolobus solfataricus菌株IT3的质粒pIT3编码的三功能单体引发酶-聚合酶结构域。将包含残基31-245（即Rep245）的pIT3复制蛋白的N末端结构域建模到古细菌质粒pRN1的双功能引发酶-聚合酶结构域的晶体结构上，并通过溶液中的分子动力学进行精制。在大肠杆菌中过量表达后，纯化Rep245蛋白，并对其核酸合成活性进行表征。描述了聚合酶活性的生化特性，例如pH，最适温度和二价阳离子金属依赖性。 Rep245能够利用核糖核苷酸和脱氧核糖核苷酸进行从头引物合成，并以模板依赖性方式合成了长达几kb的DNA产物。有趣的是，Rep245 primase-聚合酶结构域还具有末端核苷酸转移酶活性，能够以非模板方式延长合成寡核苷酸的3'-末端。对建模的Rep245结构域与其他古细菌引发酶-聚合酶的比较序列结构分析表明，一些独特的特征可以解释该结构域表现出的多方面活动。据我们所知，Rep245代表的是具有DNA和RNA合成以及末端转移酶活性的克雷纳丘菌质粒中最短的功能域。

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