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首页> 外文期刊>Journal of bacteriology >YqfS from Bacillus subtilis Is a Spore Protein and a New Functional Member of the Type IV Apurinic/Apyrimidinic-Endonuclease Family
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YqfS from Bacillus subtilis Is a Spore Protein and a New Functional Member of the Type IV Apurinic/Apyrimidinic-Endonuclease Family

机译：来自枯草芽孢杆菌的YqfS是一种孢子蛋白，是IV型Apurinic / Apyrimidinic-Endonuclease家族的新功能成员

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The enzymatic properties and the physiological function of the type IV apurinic/apyrimidinic (AP)-endonuclease homolog of Bacillus subtilis, encoded by yqfS, a gene specifically expressed in spores, were studied here. To this end, a recombinant YqfS protein containing an N-terminal His6 tag was synthesized in Escherichia coli and purified to homogeneity. An anti-His6-YqfS polyclonal antibody exclusively localized YqfS in cell extracts prepared from B. subtilis spores. The His6-YqfS protein demonstrated enzymatic properties characteristic of the type IV family of DNA repair enzymes, such as AP-endonucleases and 3′-phosphatases. However, the purified protein lacked both 5′-phosphatase and exonuclease III activities. YqfS showed not only a high level of amino acid identity with E. coli Nfo but also a high resistance to inactivation by EDTA, in the presence of DNA containing AP sites (AP-DNA). These results suggest that YqfS possesses a trinuclear Zn center in which the three metal atoms are intimately coordinated by nine conserved basic residues and two water molecules. Electrophoretic mobility shift assays demonstrated that YqfS possesses structural properties that permit it to bind and scan undamaged DNA as well as to strongly interact with AP-DNA. The ability of yqfS to genetically complement the DNA repair deficiency of an E. coli mutant lacking the major AP-endonucleases Nfo and exonuclease III strongly suggests that its product confers protection to cells against the deleterious effects of oxidative promoters and alkylating agents. Thus, we conclude that YqfS of B. subtilis is a spore-specific protein that has structural and enzymatic properties required to participate in the repair of AP sites and 3′ blocking groups of DNA generated during both spore dormancy and germination.
机译：由枯草芽孢杆菌编码的 yqfS 是一种在孢子中特异性表达的基因，其IV型嘌呤/嘧啶（AP）-核酸内切酶同系物的酶学特性和生理功能是在这里学习。为此，在大肠杆菌中合成了含有N末端His 6 标签的重组YqfS蛋白，并纯化至同质。一种抗His 6 -YqfS多克隆抗体，其在 B 制备的细胞提取物中专门定位于YqfS。 枯草孢子。 His 6 -YqfS蛋白表现出DNA修复酶IV型家族的酶学特性，如AP内切核酸酶和3'-磷酸酶。但是，纯化的蛋白质缺乏5'-磷酸酶和核酸外切酶III活性。 YqfS不仅显示出与 E的高度氨基酸同一性。在含有AP位点的DNA（AP-DNA）的存在下，大肠杆菌Nfo也具有很高的抗EDTA灭活能力。这些结果表明，YqfS具有一个三核的Zn中心，其中的三个金属原子被9个保守的碱性残基和2个水分子紧密地配位。电泳迁移率迁移分析表明，YqfS具有使其能够结合和扫描未损坏的DNA以及与AP-DNA强烈相互作用的结构特性。 yqfS 从基因上补充 E的DNA修复缺陷的能力。缺少主要的AP内切核酸酶Nfo和外切核酸酶III的大肠杆菌突变体强烈表明，其产物赋予细胞保护以抵抗氧化启动子和烷基化剂的有害作用。因此，我们得出 B的YqfS的结论。枯草芽孢杆菌是一种孢子特异性蛋白，具有参与孢子休眠和萌发过程中产生的AP位点和3'封闭DNA的修复基团所需的结构和酶学特性。

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