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首页> 外文期刊>Biochemistry >Kinetic Basis of Nucleotide Selection Employed by a Protein Template-Dependent DNA Polymerase
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Kinetic Basis of Nucleotide Selection Employed by a Protein Template-Dependent DNA Polymerase

机译：蛋白质模板依赖性DNA聚合酶采用核苷酸选择的动力学基础

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摘要

Rev 1 , a Y-family DNA polymerase, contributes to spontaneous and DNA damage-induced mutagenic events. In this paper, we have employed pre-steady-state kinetic methodology to establish a kinetic basis for nucleotide selection by human Rev!, a unique nucleotidyl transferase that uses a protein templatedirected mechanism to preferentially instruct dCTP incorporation. This work demonstrated that the high incorporation efficiency of dCTP is dependent on both substrates: an incoming dCTP and a templating base dG. The extremely low base substitution fidelity of human Revl (10~0 to 10~(-5)) was due to the preferred misincorporation of dCTP with templating bases dA, dT, and dC over correct dNTPs. Using non-natural nucleotide analogues, we showed that hydrogen bonding interactions between residue R357 of human Revl and an incoming dNTP are not essential for DNA synthesis. Lastly, human Rev I discriminates between ribonucleotides and deoxyribonucleotides mainly by reducing the rate of incorporation, and the sugar selectivity of human Revl is sensitive to both the size and orientation of the 2'-substituent of a ribonucleotide.
机译：Rev 1，Y家族DNA聚合酶，有助于自发和DNA损伤诱导的诱变事件。在本文中，我们采用了预稳态的动力学方法，为人类Rev的核苷酸选择来建立动力学基础！，一种独特的核苷酸转移酶，其使用蛋白质模板化的机制优先指示DCTP掺入。这项工作表明，DCTP的高掺入效率取决于两个基板：进入的DCTP和模板基础DG。人Revl的极低基础取代保真度（10〜0至10〜（-5））是由于DCTP的优选MISCloration，具有模板碱基DA，DT和DC在正确的DNTPS上。使用非天然核苷酸类似物，我们表明，人Revl的残基R357与进入的DNTP之间的氢键相互作用对DNA合成不是必需的。最后，通过降低掺入速率，人类Revi主要通过降低掺入速率而歧视核糖核苷酸和脱氧核糖核苷酸，并且人Revl的糖选择性对核糖核苷酸的2'-取代基的尺寸和取向敏感。

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  • 来源
    《Biochemistry》 |2010年第26期|共7页
  • 作者

    Jessica A. Brown; Jason D. Fowler; Zucai Suo;

  • 作者单位

    Department of Biochentistry The Ohio State University Columbus Ohio 43210;

    Department of Biochentistry The Ohio State University Columbus Ohio 43210;

    Department of Biochentistry The Ohio State University Columbus Ohio 43210;

  • 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

    Basis; Selection; Protein;

    机译：基础;选择;蛋白质;

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