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首页> 外文期刊>Biochemistry >Hinge Residue Ile260 of DNA Polymerase j3 Is Important for Enzyme Activity and Fidelity
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Hinge Residue Ile260 of DNA Polymerase j3 Is Important for Enzyme Activity and Fidelity

机译：DNA聚合酶J3的铰链残留髂骨260对于酶活性和保真性很重要

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摘要

DNA polymerases ensure efficient insertion of the correct dNTP into the DNA substrate.They have evolved mechanisms for discriminating among very similar dNTP substrates.DNA polymerase beta is a repair polymerase that provides a model system for a direct study of insertion fidelity.In this study,we examined the role of hinge residue Ile260 of the rat Polbeta on enzyme activity and accuracy.We changed residue 1260 to every other amino acid residue and used genetic screens to assess the activity and fidelity of the resulting mutants.The I260D,-E,-K,-N,and-R mutants are significantly less active than wild-type Polbeta.Interestingly,I260H and I260Q are active but exhibit mutator activity.This suggests that the nonpolar nature of residue 260 is important for maintaining the activity and fidelity of Polbeta.We employ molecular modeling as an aid in explaining the observed phenotypes and propose a mechanism whereby the positioning of the DNA substrate in the enzyme and within the surface of the hinge may be a key player in forming an optimal active site for phosphodiester bond formation between Watson-Crick base pairs.
机译：DNA聚合酶确保将正确的DNTP的有效插入DNA底物。它们具有用于区分非常相似的DNTP底物.DNA聚合酶β是一种修复聚合酶，其提供用于直接研究插入保真度的模型系统。这项研究，我们检查了大鼠脊粒子铰链残留物ILE260对酶活性和精度的作用。我们将残留物1260改为其他氨基酸残基，并使用遗传筛来评估所得突变体的活性和保真度。I260d，-e， - K，-N和-R突变体显着低于野生型POLBETA。互联网，I260H和I260Q是活性的，但表现出致毒剂活性。这表明残留物260的非极性性质对于维持POLBETA的活动和保真度是重要的。我们采用分子建模作为解释所观察到的表型，并提出一种机制，由此在酶和表面的表面内定位DNA底物E Hinge可以是在Watson-Crick碱基对之间形成最佳活性位点的关键球员，用于磷酸二酯键形成。

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  • 来源
    《Biochemistry》 |2005年第10期|共10页
  • 作者

    Daniela Starcevic; Shibani Dalai; Joann Sweasy;

  • 作者单位

    Departments of Genetics and Therapeutic Radiology and Program in Microbiology Yale University School of Medicine New Haven Connecticut 06520;

    Departments of Genetics and Therapeutic Radiology and Program in Microbiology Yale University School of Medicine New Haven Connecticut 06520;

    Departments of Genetics and Therapeutic Radiology and Program in Microbiology Yale University School of Medicine New Haven Connecticut 06520;

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  • 正文语种 eng
  • 中图分类 生物化学;
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