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首页> 外文期刊>Journal of Molecular Biology >Structural Mechanisms of Cooperative DNA Binding by Bacterial Single-Stranded DNA-Binding Proteins
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Structural Mechanisms of Cooperative DNA Binding by Bacterial Single-Stranded DNA-Binding Proteins

机译：基因单链DNA结合蛋白合作DNA结合的结构机制

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摘要

Bacteria encode homooligomeric single-stranded (ss) DNA-binding proteins (SSBs) that coat and protect ssDNA intermediates formed during genome maintenance reactions. The prototypical Escherichia coli SSB tetramer can bind ssDNA using multiple modes that differ by the number of bases bound per tetramer and the magnitude of the binding cooperativity. Our understanding of the mechanisms underlying cooperative ssDNA binding by SSBs has been hampered by the limited amount of structural information available for interfaces that link adjacent SSB proteins on ssDNA. Here we present a crystal structure of Bacillus subtilis SsbA bound to ssDNA. The structure resolves SsbA tetramers joined together by a ssDNA "bridge" and identifies an interface, termed the "bridge interface," that links adjacent SSB tetramers through an evolutionarily conserved surface near the ssDNA-binding site. E. coli SSB variants with altered bridge interface residues bind ssDNA with reduced cooperativity and with an altered distribution of DNA binding modes. These variants are also more readily displaced from ssDNA by RecA than wild-type SSB. In spite of these biochemical differences, each variant is able to complement deletion of the ssb gene in E. coli. Together our data suggest a model in which the bridge interface contributes to cooperative ssDNA binding and SSB function but that destabilization of the bridge interface is tolerated in cells. (C) 2018 Elsevier Ltd. All rights reserved.
机译：细菌编码HomoOligomeric单链（SS）DNA结合蛋白（SSBS），其涂覆并保护在基因组维持反应期间形成的SSDNA中间体。原型大肠杆菌SSB四聚体可以使用多种模式粘合SSDNA，其不同的模式不同的碱基的碱基的数量和结合合作率的大小。我们对由SSBS的合作SSDNA结合的机制的理解已经受到可用于链接在SSDNA上相邻SSB蛋白的接口的结构信息有限的结构信息。在这里，我们呈现枯草芽孢杆菌SSBA的晶体结构与SSDNA结合。该结构通过SSDNA“桥梁”分辨得到的SSBA四聚体，并识别界面，称为“桥接界面”，该接口将相邻的SSB四聚体通过SSDNA结合位点附近的进化水保守的表面连接。大肠杆菌SSB变体具有改变的桥接界面残留物，残留的SSDNA减少合作，并且具有改变的DNA结合模式的分布。这些变体也比野生型SSB从SSDNA置换更容易移位。尽管有这些生化差异，但每个变体能够在大肠杆菌中补充SSB基因的缺失。我们的数据组合在一起，其中桥接界面有助于协作SSDNA绑定和SSB功能，但是在细胞中容忍桥接界面的破坏化。 （c）2018年elestvier有限公司保留所有权利。

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  • 来源
    《Journal of Molecular Biology》 |2019年第2期|共18页
  • 作者

    Dubiel Katarzyna; Myers Angela R.; Kozlov Alexander G.; Yang Olivia; Zhang Jichuan; Ha Taekjip; Lohman Timothy M.; Keck James L.;

  • 作者单位

    Univ Wisconsin Sch Med &

    Publ Hlth Dept Biomol Chem 1135 Biochem Sci Bldg 420 Henry Mall;

    Univ Wisconsin Sch Med &

    Publ Hlth Dept Biomol Chem 1135 Biochem Sci Bldg 420 Henry Mall;

    Washington Univ Sch Med Dept Biochem &

    Mol Biophys 660 South Euclid Ave St Louis MO 63110 USA;

    Johns Hopkins Univ Dept Biophys &

    Biophys Chem 725 N Wolfe St Baltimore MD 21205 USA;

    Johns Hopkins Univ Dept Biophys &

    Biophys Chem 725 N Wolfe St Baltimore MD 21205 USA;

    Johns Hopkins Univ Dept Biophys &

    Biophys Chem 725 N Wolfe St Baltimore MD 21205 USA;

    Washington Univ Sch Med Dept Biochem &

    Mol Biophys 660 South Euclid Ave St Louis MO 63110 USA;

    Univ Wisconsin Sch Med &

    Publ Hlth Dept Biomol Chem 1135 Biochem Sci Bldg 420 Henry Mall;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子生物学;
  • 关键词

    DNA replication; DNA repair; protein-DNA interactions; SSB; RecA;

    机译：DNA复制;DNA修复;蛋白质-DNA相互作用;SSB;RECA;

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